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ObjectiveTo investigate the regulatory effect of Danggui Shaoyaosan on adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR)/Unc-51-like kinase-1 (ULK1) signaling pathway in the rat model of metabolism-associated fatty liver disease (MAFLD). MethodSixty SD rats were randomized into control, model, western medicine (polyene phosphatidylcholine capsules,0.144 g·kg-1), and low-, medium-, and high-dose (2.44, 4.88, 9.76 g·kg-1, respectively) Danggui Shaoyaosan groups. After being fed with a high-fat diet for 8 weeks, the rats in each group were administrated with corresponding drugs for 4 weeks. At the end of drug treatment, serum and liver tissue were collected for subsequent determination of related indicators. ResultCompared with the control group, the model group showed increased contents of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum, increased contents of TC, TG, and free fatty acids (FFAs) in the liver (P<0.01), and decreased content of high-density lipoprotein cholesterol (HDL-C) in the serum (P<0.01). Furthermore, the model group showed down-regulated protein levels of p-AMPK, microtubule-associated protein 1 light chain 3B (LC3B) Ⅱ, Beclin1, and ULK1 (P<0.01) and up-regulated protein levels of p-mTOR and ubiquitin-binding protein p62 in the liver (P<0.01). The hepatic steatosis was obvious and the NAFLD activity score (NAS) and oil red O staining area increased in the model group, (P<0.05, P<0.01). Compared with the model group, Danggui Shaoyaosan reduced the contents of TC and TG and the activities of ALT and AST in the serum, lowered the levels of TC, TG, and FFA in the liver, down-regulated the protein levels of p-mTOR and p62 (P<0.01), elevated the serum HDL-C level, and up-regulated the protein levels of p-AMPK, LCBⅡ, Beclin1, and ULK1 in the liver (P<0.05, P<0.01). Moreover, it alleviated hepatic steatosis and decreased the NAS and oil red O staining area (P<0.05, P<0.01). ConclusionDanggui Shaoyaosan has therapeutic effect on MAFLD rats by regulating AMPK/mTOR/ULK1 signaling pathway to enhance autophagy.
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ObjectiveTo observe the effects of Hirudo, Notoginseng Radix et Rhizoma, and drug pair on renal pathological morphology and protein phosphatase 2A (PP2A)/adenylate activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signal pathway in rats with chronic renal failure (CRF). MethodThe 55 male SD rats were randomly divided into a normal group (n=11) and a modeling group (n=44). The normal group was fed conventionally, and the modeling group was given 0.25 g·kg-1·d-1 adenine by gavage for 28 days to replicate the CRF model. After successful modeling, rats were randomly divided into model group, Hirudo group (3 g·kg-1·d-1), Notoginseng Radix et Rhizoma group (3 g·kg-1·d-1), and Hirudo + Notoginseng Radix et Rhizoma group (3 g·kg-1·d-1), with 9 rats in each group. The normal group and model group were given a constant volume of normal saline by intragastric administration for 30 days. At the end of the experiment, the levels of serum creatinine (SCr) and urea nitrogen (BUN) in all groups were measured. The renal pathological morphology changes were observed by hematoxylin-eosin (HE) staining, Masson staining, and electron microscopy. The mRNA expressions of PP2A, AMPK, and mTOR were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). The protein expression levels of PP2A, AMPK, phosphorylation(p)-AMPK, mTOR, and p-mTOR in renal tissue were detected by Western blot. ResultCompared with the normal group, the renal pathological structure changes were obvious, and the levels of SCr and BUN were significantly increased. The mRNA expression of PP2A, protein expression of PP2A, and p-mTOR/mTOR expression were significantly increased, and the p-AMPK/AMPK was significantly decreased in the model group (P<0.05). Compared with the model group, the renal pathological morphology changes were significantly improved, and the levels of SCr and BUN were significantly decreased. The mRNA expression of PP2A, protein expression of PP2A, and p-mTOR/mTOR expression in the renal tissue were significantly decreased, and the p-AMPK/AMPK was significantly increased (P<0.05) in all groups after drug intervention. In addition, the effect in the Hirudo+Notoginseng Radix et Rhizoma group was better. The mRNA expression levels of AMPK and mTOR in the renal tissue were not significantly different among the normal group, model group, and other groups. ConclusionThe efficacy of Hirudo and Notoginseng Radix et Rhizoma pairs in improving renal fibrosis in rats with CRF is significantly better than that of the single drug, and its improvement on renal fibrosis in rats with CRF may be related to the regulation of PP2A/AMPK/mTOR signaling pathway.
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ObjectiveTo explore the protective mechanism of paeoniflorin on mice with ulcerative colitis (UC) through the adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) autophagy pathway. MethodUC mouse model was established by allowing mice freely drink 4% DSS, and 56 BALB/c male mice were randomly divided into model group, AMPK inhibitor group (20 mg·kg-1), paeoniflorin (50 mg·kg-1) + inhibitor (20 mg·kg-1) group, and high dose (50 mg·kg-1), medium dose (25 mg·kg-1), and low dose (12.5 mg·kg-1) paeoniflorin groups. After seven days of drug intervention, the protective effect of paeoniflorin on mice with UC was determined by comparing the body weight, disease activity index (DAI) changes, and Hematoxylin-eosin (HE) staining results. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the serum of mice in each group, and immunofluorescence was utilized to detect microtubule-associated protein 1 light chain 3 (LC3) content in the colon, AMPK, mTOR proteins, and their phosphorylated proteins including p-AMPK and p-mTOR in the colon tissue were detected by Western blot, and the mRNA expression levels of AMPK, mTOR, Beclin1, LC3, and p62 were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultCompared with the blank group, the model group showed a decrease in body mass, an increase in DAI score, and severe pathological damage to the colon. The levels of inflammatory factors including TNF-α and IL-6 increased in serum (P<0.01), while the protein levels of LC3 and p-AMPK/AMPK were down-regulated in colon tissue, and those of p-mTOR/mTOR were up-regulated (P<0.01). The mRNA expression levels of AMPK and LC3 were down-regulated, while the mRNA expression levels of mTOR and p62 were up-regulated (P<0.01). Compared with the model group and the paeoniflorin + inhibitor group, the mice treated with paeoniflorin showed an increase in body mass, a decrease in DAI score, a reduction in pathological damage to colon tissue, and a reduction in the levels of inflammatory factors of TNF-α and IL-6 in serum (P<0.05). The protein levels of LC3 and p-AMPK/AMPK in colon tissue were up-regulated, while the protein levels of p-mTOR/mTOR were down-regulated (P<0.01). The mRNA expression levels of AMPK, Beclin1, and LC3 were up-regulated, while the mRNA expression of mTOR and p62 were down-regulated (P<0.01). The colon tissue of the inhibitor group was severely damaged, and the trend of various indicators was completely opposite to that of the high dose paeoniflorin group. ConclusionPaeoniflorin can enhance autophagy and reduce inflammatory damage in mice with UC by activating the AMPK/mTOR signaling pathway and thus play a protective role.
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Solid organ transplantation has significantly prolonged the survival of patients with end-stage diseases. However, long-term use of immunosuppressants will increase the risk of post-transplantation diabetes mellitus (PTDM) in the recipients, thereby elevating the risk of infection, cardiovascular disease and death. In recent years, with persistent improvement of diagnostic criteria of PTDM, clinicians have deepened the understanding of this disease. Compared with type 2 diabetes mellitus, PTDM significantly differs in pathophysiological characteristics and clinical progression. Hence, different treatment strategies should be adopted. Early identification of risk factors of organ transplant recipients, early diagnosis and intervention are of significance for improving the quality of life of recipients, prolonging the survival of grafts and reducing the fatality of recipients. Therefore, the diagnosis, incidence and risk factors of PTDM were reviewed in this article, aiming to provide reference for clinicians to deliver prompt diagnosis and intervention for PTDM.
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Ischemic stroke (IS) is a serious cerebrovascular disease common in clinical practice. Targeting the pathogenesis of IS, intravenous thrombolysis for restoring blood flow is still the most effective therapy. However, intravenous thrombolysis has shortcomings such as increased bleeding risk, narrow therapeutic window, and contraindications, which limited its clinical application. Protection of the ischemic brain tissue before full recovery of blood flow is associated with the prognosis of IS. Studies have identified multiple pathways in the alleviation of the brain injury caused by IS, such as the mammalian target of rapamycin (mTOR) signaling pathway. Traditional Chinese medicine (TCM) has abundant therapies and unique advantages in the treatment of IS, especially in alleviating symptoms and improving the quality of life of patients. After the onset of IS, TCM can be integrated with Western medicine to play a role in the whole process of treatment, rehabilitation, and recurrence prevention as soon as possible, thus maximizing patient benefits. TCM has clinical significance for the recovery of neurological and motor functions after IS. Studies have shown that TCM can reduce the cerebral injury caused by IS by regulating and activating the mTOR signaling pathway, thereby regulating autophagy, inhibiting apoptosis of nerve cells, and reducing oxidative stress and inflammation. TCM exerts a positive effect for achieving cerebral protection and improving the prognosis of IS and provides new ideas for the prevention and treatment of IS. This article introduces the role of the mTOR signaling pathway in the pathogenesis of IS and reviews the research progress in the TCM regulation of this pathway in the treatment of IS, aiming to provide new therapeutic ideas and systematic scientific reference for the treatment of IS with TCM.
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Objective To explore the mechanism of malignant behavior of cervical cancer(CC)cells based on AMP-activated protein kinase(AMPK)/rapamycin target protein(mTOR)signaling pathway mediated by nucleo-tide-binding oligomerization domain receptor 2(NOD2).Methods Bioinformatics analysis was performed to deter-mine the expression of NOD2 in CC tissue.Plasmids targeting NOD2(shNOD2)and shRNAs negative control(shNC),NOD2 overexpression(NOD2)and vectors(Vec)were transfected into CC cells.The effect of NOD2 on the growth of CC cells was determined by cell counting kit-8 assay,colony formation and Transwell cell invasion as-say.Transcriptome analysis was performed by high throughput RNA sequencing(RNA-Seq).Western blot was used to detect the expression of NOD2,AMPK/mTOR signaling pathway and autophagy protein in the cell line.24 female BALB/c nude mice were randomly divided into four groups,with 6 mice in each group:vector group(Vec group),NOD2 overexpression group(NOD2 group),shNC group and shNOD2 group.The distant metastasis mod-el was established in mice,and the fluorescence intensity of lung metastasis was monitored and the number of lung metastasis nodules was counted.Results On-line database analysis showed that the expression of NOD2 in CC tis-sues was significantly higher than that in normal tissues,and there were significant differences in the mRNA expres-sion of NOD2 in different stages of CC(P<0.05).In addition,the high expression of NOD2 was associated with poor overall survival and disease-free survival(P<0.05).NOD2 overexpression promoted the proliferation,colony formation,migration and invasion of CC cells,while NOD2 knock-down was the opposite.Consistent with the re-sults in vitro,the lung colonization and lung metastasis of CC cells in NOD2 group were significantly higher than those in Vec group(P<0.05),while those in shNOD2 group were significantly lower than those in shNC group(P<0.05).RNA-Seq results showed that the expression of NOD2 was significantly related to AMPK signal activa-tion,mTOR signal inhibition,autophagy regulation pathway activation and autophagy formation.Compared with shNC group,the expression of phosphorylated AMPK and LC3 protein decreased significantly in shNOD2 group(P<0.05),and the expression levels of phosphorylated mTOR and p62 protein increased significantly(P<0.05).Compared with Vec group,the expression levels of LC3 and AMPK protein in NOD2 group increased significantly(P<0.05),and the expression levels of phosphorylated mTOR and p62 protein decreased significantly(P<0.05).Compared with shNC group,the point accumulation of GFP-mRFP-LC3 in shNOD2 group decreased signifi-cantly(P<0.05).Compared with Vec group,the point accumulation of GFP-mRFP-LC3 increased significantly(P<0.05).Conclusion NOD2 may promote the proliferation,migration and invasion of CC through AMPK/mTOR signal,and its mechanism partly involves autophagy activation.
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Objective:To discuss the effect of downregulating of high mobility group box protein 2(HMGB2)expression on the biological behavior of the liver cancer cells and the epithelial-mesenchymal transition(EMT)process,and to clarify its mechanism.Methods:The human liver cancer LM3 cells at logarithmic growth phase were divided into negative control group and HMGB2 RNA interference group(HMGB2 siRNA group);the cells in two groups were transfected with RNA oligonucleotides(RNA oligos)with irrelevant sequences and RNA oligos designed to knock down HMGB2,and the Lipofectamine 2000 was regarded as the vector.The expression levels of HMGB2 mRNA and protein in the cells in two groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods;cell scratch assay and Transwell chamber assay were used to detect the migration and invasion abilities of the cells in two groups;the expression levels of E-cadherin,N-cadherin,and Vimentin proteins and protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway related proteins in the cells in two groups were detected by Western blotting method.Results:Compared with negative control group,the expression levels of HMGB2 mRNA and protein in the cells in HMGB2 siRNA group were significantly decreased(P<0.05),the cell scratch healing rate was significantly decreased(P<0.01),the number of invasion cells was significantly decreased(P<0.01),and the expression level of E-cadherin protein in the cells was significantly increased(P<0.01),while the expression levels of N-cadherin,Vimentin,mTOR,AKT,and phosphorylated AKT(p-AKT)proteins in the cells were significantly decreased(P<0.05 or P<0.01).Conclusion:Downregulating the expression of HMGB2 can reduce the migration and invasion abilities of the liver cancer LM3 cells and inhibit the EMT,and its mechanism may be related to regulating the expression of the AKT/mTOR pathway related proteins.
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Objective To investigate the clinical effect of rapamycin-eluting vertebral artery stent in the treatment of severe ostial vertebral artery stenosis(OV AS),and to analyze the incidence of postoperative in-stent restenosis(ISR).Methods A total of 96 patients with severe OVAS,who received stenting angioplasty at authors'hospital between November 2020 and May 2022,were retrospectively collected.The patients were divided into the observation group(n=48)and the control group(n=48).For the patients of the observation group implantation of rapamycin-eluting vertebral artery stent was carried out,while for the patients of the control group implantation of peripheral balloon dilatation bare metal stent(BMS)was performed.The perioperative basic data,the incidence of complications during follow-up period,and the postoperative incidence of ISR were compared between the two groups.Results Successful stent implantation was achieved in all patients of both groups.During perioperative period no complications such as transient ischemia attack(TIA),dropping-off or fracture of the stent,vertebral artery or stent-related stroke occurred.No statistically significant differences in the length and the diameter of the implanted stents,in the preoperative vertebral artery stenosis ratio,and in the postoperative residual stenosis ratio existed between the two groups(all P>0.05).In both groups,the postoperative residual stenosis ratio was<20%.The patients were followed up for a mean period of(12.33±5.82)months(range of 6-18 months),the incidence of postoperative vertebral artery or stent-related stroke in the observation group and the control group was 0%and 4.17%respectively,the difference between the two groups was not statistically significant(P>0.05).The improvement of clinical symptoms such as dizziness,vertigo,etc.was observed in 47 patients of the observation group and in 45 patients of the control group,and no recurrent posterior circulation TIA or stent-related thrombotic event occurred.The incidence of postoperative restenosis in the observation group was 10.42%,which was significantly lower than 29.17%in the control group(P<0.05).Conclusion Rapamycin-eluting vertebral artery stent can safely and effectively treat severe OVAS and reduce the incidence of postoperative ISR.(J Intervent Radiol,2024,33:275-279)
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Objective:To investigate the influences of lupinol on the proliferation,apoptosis and invasion of cer-vical cancer cells by regulating autophagy mediated by phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway.Methods:The proliferation rate of human cervical cancer cell line HeLa cells treated with 0,10,25,50,70,90 μmol/L lupinol was determined,and the appropriate concentration of lupinol was screened out.HeLa cells cultured in vitro were randomly grouped into control group,low-dose lupinol group,high-dose lupinol group,740 Y-P group(PI3K activator),and high-dose lupinol+740 Y-P group.After group intervention with lupinol and 740 Y-P,MDC fluorescence staining was used to detect the forma-tion of autophagic vacuolation of HeLa cells in each group;western blot was used to detect the expression of au-tophagy and PI3K/AKT/mTOR pathway-related proteins in HeLa cells in each group.HeLa cells cultured in vitro were randomly grouped into control group,low-dose lupinol group,high-dose lupinol group,high-dose lupinol+rapamycin(Rapa),and high-dose lupinol+3-methyladenine(3-MA)group.After the intervention of high dose of lupinol,Rapa and 3-MA,the proliferation of HeLa cells in each group was detected by MTT assay and plate colony formation assay;flow cytometry was used to detect the apoptosis of HeLa cells in each group;transwell assay was used to detect the invasion of HeLa cells in each group;western blot was used to detect the expressions of proliferation,apoptosis and epithelial-mesenchymal transition-related proteins in HeLa cells in each group.Re-sults:Compared with the control group,the relative content of autophagic vacuoles,the protein expressions of Mi-crotubule-associated protein 1A/1 B-light chain 3(LC3)Ⅱ/LC3Ⅰ,and Beclin-1 in the low and high dose lupinol groups were all increased(P<0.05),the phosphorylated PI3K(p-PI3K)/PI3K,phosphorylated AKT(p-AKT)/AKT,and phosphorylated mTOR(p-mTOR)/mTOR decreased(P<0.05);the relative content of autophagic vac-uoles,the protein expressions of LC3Ⅱ/LC3Ⅰ,and Beclin-1 in the high-dose lupinol group were further increased compared with the low-dose lupinol group(P<0.05),the p-PI3K/PI3K,p-AKT/AKT,and p-mTOR/mTOR were further decreased(P<0.05);the relative content of autophagic vacuoles,the protein expressions of LC3Ⅱ/LC3Ⅰ,and Beclin-1 in 740 Y-P group decreased compared with the control group(P<0.05),the p-PI3K/PI3K,p-AKT/AKT,and p-mTOR/mTOR increased(P<0.05).Compared with the high-dose lupinol group,the relative content of autophagic vacuoles,the protein expressions of LC3Ⅱ/LC3Ⅰ,and Beclin-1 in the high-dose lupinol+740 Y-P group decreased(P<0.05),the p-PI3K/PI3K,p-AKT/AKT,and p-mTOR/mTOR increased(P<0.05).Com-pared with the control group,the cell proliferation rate,colony formation rate,invasion number,and the protein ex-pressions of proliferating cell nuclear antigen(PCNA),B cell lymphoma 2(Bcl-2)and Vimentin in the low and high dose groups of lupinol were all decreased(P<0.05),the apoptosis rate,and the protein expressions of Bcl-2 as-sociated x protein(Bax)and zonula occludens protein 1(ZO-1)were all increased(P<0.05);compared with the low-dose lupinol group,the cell proliferation rate,colony formation rate,invasion number,and the protein expres-sions of PCNA,Bcl-2 and Vimentin in the high-dose lupinol group were further decreased(P<0.05),the apopto-sis rate,and the protein expressions of Bax and ZO-1 were further increased(P<0.05).Compared with the high-dose lupinol group,the cell proliferation rate,colony formation rate,invasion number,and the protein expres-sions of PCNA,Bcl-2 and Vimentin in the high-dose lupinol+Rapa group were increased(P<0.05),the apopto-sis rate,and the protein expressions of Bax and ZO-1 were decreased(P<0.05);the cell proliferation rate,colo-ny formation rate,invasion number,and the protein expressions of PCNA,Bcl-2 and Vimentin in the high-dose lu-pinol+3-MA group were decreased(P<0.05),the apoptosis rate,and the protein expressions of Bax and ZO-1 were increased(P<0.05).Conclusions:Lupinol induces protective autophagy by inhibiting the PI3K/AKT/mTOR pathway,thereby promoting the apoptosis of cervical cancer cells and inhibiting their proliferation and inva-sion.Activation of autophagy attenuates the effects of lupinol on the proliferation,apoptosis and invasion of cervi-cal cancer cells.
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BACKGROUND:In recent years,it has been found that some traditional Chinese medicine monomers can alleviate oxidative stress and apoptosis of the skin flap,promote vascular regeneration of the skin flap and prevent skin flap necrosis by activating autophagy. OBJECTIVE:To review the research progress of traditional Chinese medicine monomer regulating autophagy in preventing flap necrosis. METHODS:The Chinese and English key words were"traditional Chinese medicine(TCM),autophagy,skin flaps".The first author searched the relevant articles published in CNKI and PubMed databases from January 2010 to October 2022.A total of 196 articles were retrieved in the preliminary screening and then screened according to the inclusion and exclusion criteria.The quality assessment was conducted by reading the literature titles and abstracts.Finally,55 articles were summarized. RESULTS AND CONCLUSION:(1)The regulation of autophagy is mediated by AMPK/mTOR,PI3K/AKT and other signaling pathways.Activation of autophagy can alleviate the oxidative stress and apoptosis of the flap,promote the regeneration of blood vessels in the flap,and prevent flap necrosis.(2)Terpenoids(Betulinic acid,Andrographolide,Notoginseng Triterpenes,Catalpa),phenolic compounds(Resveratrol,Curcumin,Gastrodin),phenolic acids(Salvianolic acid B)and steroid compounds(Pseudoginsenoside F11)in traditional Chinese medicine monomers can alleviate oxidative stress and apoptosis of skin flap by regulating related signaling pathways to activate autophagy,promote skin flap angiogenesis and promote skin flap survival.(3)Studying the research progress of traditional Chinese medicine monomer to prevent flap necrosis by regulating autophagy can provide a reference and theoretical basis for traditional Chinese medicine to prevent flap necrosis and promote flap healing in the clinic.
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Objective To investigate the effect of atractylenolideⅠon lung injury in rats with recurrent respiratory tract infection(RRTI)of lung and spleen qi deficiency by regulating phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)pathway.Methods Eighty-four rats were randomly separated into a control group,a model group,a low-dose atractylenolideⅠgroup,a high-dose atractylenolideⅠgroup,a positive drug group,an insulin-like growth factor-1(IGF-1)group,and a high-dose atractylenolide Ⅰ+IGF-1 group,with 12 rats in each group.Except for the control group,the RRTI rat model of lung and spleen qi deficiency was constructed using a combination of fatigue,dietary disorders,and fumigation method with shavings and tobacco among rats in other groups.After the model is successfully copied,the model was administered once a day for 6 weeks.Animal lung function instrument was applied to detect the changes of peak expiratory flow(PEF),forced expiratory volume in first second(FEV1),forced vital capacity(FVC)in rats.The changes of wet/dry mass ratio of lungs in rats were detected.HE staining was applied to detect pathological changes of lung tissue in rats of each group.ELISA was applied to detect the levels of interleukin(IL)-6,tumor necrosis factor-α(TNF-α),malondialdehyde(MDA)and the activity of superoxide dismutase(SOD)in rat lung tissue.Western Blot was applied to determine the protein expressions of p-PI3K,p-Akt,and p-mTOR in rat lung tissue.Results Compared with the control group,rats in the model group showed a decrease in PEF,FEV1 and FVC(P<0.01)and an increase in the wet/dry mass ratio of lungs(P<0.01).The alveolar septa in lung tissues had become larger.Pulmonary interstitial edema and a large amount of inflammatory cell infiltration were found.The levels of IL-6,TNF-α and MDA in lung tissue increased(P<0.01),and the SOD activity decreased(P<0.01).The protein expressions of p-PI3K,p-Akt,and p-mTOR in lung tissue increased(P<0.01).Compared with the model group,rats in the low-,high-dose atractylenolideⅠgroups,and positive drug group showed an increase in PEF,FEV1,and FVC,and a decrease in the wet/dry mass ratio of lungs(P<0.01).Pathologic damage in lung tissue was alleviated.The levels of IL-6,TNF-α,MDA decreased and SOD activity in lung tissue increased(P<0.01).The protein expressions of p-PI3K,p-Akt,and p-mTOR in lung tissue decreased(P<0.01),while the corresponding indicators in the IGF-1 group showed opposite trends(P<0.01).Compared with the high-dose group of atractylenolide I,rats in the high-dose atractylenolide I+IGF-1 group showed a decrease in PEF,FEV1 and FVC,and an increase in the wet/dry mass ratio of lungs(P<0.01).Pathologic damage in lung tissue was increased.The levels of IL-6,TNF-α,MDA increased and the SOD activity in lung tissue decreased(P<0.01).The protein expressions of p-PI3K,p-Akt,and p-mTOR in lung tissue increased(P<0.05,P<0.01).Conclusion AtractylenolideⅠmay improve lung injury in RRTI rats of lung and spleen qi deficiency by inhibiting the PI3K/Akt/mTOR pathway.
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ObjectiveTo determine the syndrome of a rat model of follicular dysplasia induced by Tripterygium glycosides based on prescriptions and investigate the mechanism of traditional Chinese medicine intervention via the adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR)/hypoxia-inducible factor-1 (HIF-1)/vascular endothelial growth factor (VEGF) pathway. MethodForty-eight rats with regular estrous cycles were randomly assigned into a normal group (n=8) and a modeling group (n=40). The rats in the modeling group were administrated with Tripterygium glycoside suspension (75 mL·kg-1) by gavage for 30 days. The modeled rats were assigned into model, Siwutang (3.69 g·kg-1), Youguiyin (3.11 g·kg-1), Zuoguiyin (7.29 g·kg-1), and Guishenwan (10.35 g·kg-1) groups, with 8 rats in each group. The drug intervention lasted for 14 days. The changes of estrous cycle were detected by Pap staining, and a stereoscope was used to observe the morphology of the ovarian tissue. Hematoxylin-eosin staining was employed to observe the pathological changes and follicle count in the ovarian tissue. Enzyme-related immunosorbent assay (ELISA) was used to measure the levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) in the serum. Real-time fluorescence quantitative polymerase chain reaction and Western blot were employed to determine the mRNA and protein levels, respectively, of AMPK, mTOR, HIF-1, and VEGF in the ovarian tissue. ResultCompared with the normal group, the model group had a disordered estrous cycle, reduced secondary and mature follicles, increased atretic follicles, elevated FSH and LH levels, lowered E2 level, up-regulated mRNA and protein levels of AMPK, and down-regulated mRNA and protein levels of mTOR, HIF-1, and VEGF (P<0.01). Compared with the model group, Guishenwan increased secondary and mature follicles, decreased atretic follicles, lowered the FSH and LH levels, elevated the E2 level, down-regulated the mRNA and protein levels of AMPK, and up-regulated the mRNA and protein levels of mTOR, HIF-1, and VEGF (P<0.01). Compared with Guishenwan group, Siwutang, Youguiyin, and Zuoguiyin decreased mature follicles, increased atretic follicles (P<0.01), elevated the LH (P<0.01) and FSH (P<0.05) levels, and lowered the E2 level (P<0.05). In addition, Youguiyin up-regulated the protein level of AMPK (P<0.05) and down-regulated the mRNA levels of mTOR and HIF-1 (P<0.01) as well as the mRNA and protein levels of VEGF (P<0.01). Siwutang down-regulated the mRNA levels of mTOR and HIF-1 as well as the mRNA and protein levels of VEGF (P<0.05). Zuoguiyin down-regulated the mRNA level of mTOR and the protein and mRNA levels of VEGF (P<0.05). ConclusionGuishenwan may improve the ovarian function and promote follicle maturation in a rat model of follicular dysplasia by inhibiting the AMPK/mTOR/HIF-1/VEGF pathway, with the therapeutic effect superior to Zuoguiyin, Youguiyin, and Siwutang. It was hypothesized that this model presented the syndrome of kidney-essence deficiency.
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ObjectiveTo investigate the effects of berbamine hydrochloride on sorafenib resistance in hepatocellular carcinoma cells and the underlying mechanisms. MethodThe sorafenib-resistant cell line SMMC-7721/S was selected by the concentration increment method starting at 1.25 μmol·L-1 sorafenib. Both SMMC-7721 and SMMC-7721/S cells were treated with 0, 2.5, 5, 10, 15, 20 μmol·L-1 sorafenib, and the cell counting kit-8 (CCK-8) assay was employed to determine the half maximal inhibitory concentration (IC50) and calculate the resistance index (RI). Western blot was conducted to compare the expression of proteins involved in autophagy and phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathway between SMMC-7721 and SMMC-7721/S cells. Furthermore, SMMC-7721/S cells were treated with 5 μmol·L-1 berbamine hydrochloride alone or in combination with 2.5, 5, 10 μmol·L-1 sorafenib, and the cell growth was assessed by the CCK-8 assay. In addition, SMMC-7721 and SMMC-7721/S cells were treated with 5 μmol·L-1 berbamine hydrochloride alone or in combination with 5 μmol·L-1 sorafenib, and the cell proliferation was examined by the colony formation assay. The immunofluorescence assays with Microtubule-associated protein 1 light chain 3 (LC3) and LysoTracker as probes were employed to assess the lysosomal acidification in SMMC-7721 cells treated with 5 μmol·L-1 berbamine hydrochloride or 0.1 μmol·L-1 autophagy inhibitor bafilomycin A1 (Baf). Further, the expression of proteins involved in autophagy and PI3K/Akt/mTOR signaling pathway was determined by Western blot and compared between groups. ResultSorafenib showed the IC50 of 9.56 mol·L-1 (P<0.01) and 7.99 mol·L-1 for SMMC-7721/S and SMMC-7721 cells, respectively, at 24 h. The resistance index (RI) of SMMC-7721/S for sorafenib was 1.20 (P<0.01), which indicated mild resistance. Compared with SMMC-7721 cells, SMMC-7721/S cells exhibited up-regulated expression of p-mTOR, p-Akt, and LC3Ⅱ, down-regulated expression of p62 protein (P<0.01), and unchanged Akt protein level. CCK-8 and colony formation assays demonstrated that the combination of berbamine hydrochloride and sorafenib exhibited a synergistic effect (Q>1.15), with berbamine hydrochloride partially reversing the resistance of liver cancer cells to sorafenib. The immunofluorescence detection of LC3 revealed that berbamine hydrochloride and Baf significantly increased LC3 in SMMC-7721 cells. The detection with LysoTracker as the probe showed that berbamine hydrochloride inhibited the acidity of lysosomes in SMMC-7721 cells (P<0.01), indicating the suppression of autophagy. Berbamine hydrochloride further enhanced the downregulation of p-mTOR and p-Akt protein levels and did not change the Akt protein level in SMMC-7721 cells exposed to sorafenib. Berbamine hydrochloride inhibited the increase in p-mTOR expression, down-regulated the p-Akt protein level, and did not change the total Akt protein level in the SMMC-7721/S cells exposed to sorafenib. ConclusionBerbamine hydrochloride can ameliorate the resistance of liver cancer cells to sorafenib by inhibiting cellular autophagy and the PI3K/Akt/mTOR signaling pathway.
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ObjectiveTo explore the intervention effect and molecular mechanism of Dabufei decoction in Dunhuang formula combined with cisplatin on Lewis lung adenocarcinoma-bearing mice. MethodFifty C57BL/6J mice were used, with 10 randomly assigned to the blank group (without modeling), and 40 subcutaneously inoculated with Lewis cells to establish a Lewis lung adenocarcinoma-bearing mouse model. These 40 mice were randomly divided into the following four groups (with 10 mice in each group): Model group (equal volume of physiological saline), cisplatin group (5 mg·kg-1), Dabufei decoction group (14.35 g·kg-1·d-1), and Dabufei decoction combined with cisplatin group (Dabufei decoction 14.35 g·kg-1·d-1 + cisplatin 5 mg·kg-1). Each group was treated continuously for 14 days. The general condition of the mice was observed, body weight changes were recorded, and the tumor inhibition rate, spleen index, and thymus index were calculated. Peripheral blood white blood cell (WBC), platelet (PLT), and hemoglobin (HGB) were detected by routine blood tests. Flow cytometry was used to detect the expression of CD4+CD25+FoxP3+ regulatory T cells (Treg) and natural killer (NK) cells in the spleen. Western blot and real-time quantitative polymerase chain reaction (Real-time PCR) were used to determine the expression of proteins and mRNA related to the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway in tumor tissues. ResultCompared with the blank group, the model group showed decreased body weight (P<0.05), spleen index, and thymus index (P<0.05), decreased percentage of NK cells in the spleen (P<0.05), increased percentage of Treg cells (P<0.05), and decreased counts of WBC, PLT, and HGB (P<0.05). Compared with the model group, the Dabufei decoction group exhibited significant tumor growth inhibition, increased body weight, and reduced tumor weight (P<0.05), increased percentage of NK cells (P<0.05), decreased proportion of Treg cells (P<0.05), and increased counts of WBC, PLT, and HGB (P<0.05). In the cisplatin group, tumor growth was significantly inhibited, body weight significantly decreased (P<0.05), and tumor weight significantly reduced (P<0.05). The spleen index and thymus index decreased (P<0.05), and the percentage of Treg cells significantly decreased (P<0.05). The counts of WBC, PLT, and HGB significantly decreased (P<0.05). In the Dabufei decoction combined with cisplatin group, tumor growth was significantly inhibited, and tumor weight significantly reduced (P<0.05). The levels of phosphorylated PI3K, Akt, and mTOR proteins and mRNA in tumor tissues were significantly reduced in all medication groups (P<0.05). Compared with the cisplatin group, the Dabufei decoction combined with cisplatin group showed significantly inhibited tumor growth, reduced tumor weight (P<0.05), increased body weight (P<0.05), increased spleen index and thymus index (P<0.05), increased percentage of NK cells (P<0.05), decreased percentage of Treg cells (P<0.05), significantly increased counts of WBC, PLT, and HGB (P<0.05), and reduced levels of phosphorylated PI3K, Akt, and mTOR and their mRNA (P<0.05). ConclusionDabufei decoction combined with cisplatin has a synergistic effect with reduced toxicity, effectively regulating immune function, increasing the proportion of NK cells, reducing the proportion of Treg cells, improving bone marrow suppression, and downregulating the PI3K/Akt/mTOR signaling pathway to inhibit tumor growth in Lewis lung adenocarcinoma-bearing mice.
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ObjectiveTo explore the therapeutic mechanism of Faeces Bombycis on diabetic gastroparesis (DGP) rats based on phosphatidylinositol 3-kinase/protein kinase B/mammalian rapamycin target protein (PI3K/Akt/mTOR) signaling pathway. MethodDGP rat model was prepared by random selection of 15 out of 105 rats as blank group. The rats successfully constructed were randomly divided into model group, high-,medium- and low- dose groups (3.2, 1.6, 0.8 g·kg-1) and moxapride group (1.5 mg·kg-1), with 12 rats in each group, and were given gavage for 4 weeks. The gastric emptying rate and random blood glucose were measured. The morphological changes of gastric antrum were observed by hematoxylin-eosin (HE) staining, and the expression of the c-Kit gene was analyzed by immunohistochemistry. The apoptosis of Cajal interstitial cells was observed by in situ end labeling (TUNEL) staining, and the protein expressions of PI3K, phosphorylation(p)-PI3K, Akt, p-Akt, mTOR, and p-mTOR were detected by Western blot. ResultCompared with the blank group, the gastric emptying rate of the model group decreased significantly (P<0.01), and the glandular structure of the gastric antrum was destroyed. The expression of c-Kit decreased (P<0.01), and the apoptosis of Cajal interstitial cells (ICC) increased. Compared with the model group, the gastric emptying rate in the high, middle, and low-dose groups of Faeces Bombycis extract and mosapride group increased significantly (P<0.01). The glandular structure of the gastric antrum became closer, and the apoptosis of ICC decreased. The expression of c-Kit in the high dose group of Faeces Bombycis extract increased significantly. After Western blot testing, compared with the blank group, the protein expression of p-Akt/Akt, p-PI3K/PI3K, and p-mTOR/mTOR in the model group increased. Compared with the model group, the protein expression of p-Akt/Akt in the high dose group of Faeces Bombycis extract decreased (P<0.01), and the protein expression of p-PI3K/PI3K decreased in the middle and low dose groups of Faeces Bombycis extract and mosapride group decreased (P<0.05, P<0.01). The protein expression of p-mTOR/mTOR decreased in the low dose group of Faeces Bombycis extract (P<0.05). In terms of random blood glucose, compared with the blank group, the random blood glucose in the model group increased significantly (P<0.01), and compared with the model group, the random blood glucose in the high and middle dose groups of Faeces Bombycis extract decreased significantly (P<0.05). Compared with mosapride group, the protein expression of p-Akt/Akt decreased in the high dose group of Faeces Bombycis extract (P<0.05), and the protein expression of p-PI3K/PI3K increased in the high, middle, and low dose groups of Faeces Bombycis extract (P<0.05, P<0.01). ConclusionFaeces Bombycis extract can increase gastric emptying rate, reduce ICC apoptosis, and lower random blood glucose in DGP rats. The high dose group of Faeces Bombycis extract has a significant effect on inhibiting ICC apoptosis, and its mechanism may be related to the regulation of the PI3K/Akt/mTOR signaling pathway.
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ObjectiveTo investigate the clinical efficacy of Qihuang Jianpi Zishen Granules in the treatment of systemic lupus erythematosus (SLE) and its effect on the signal transducer and activator of tranSCription 3/mammalian target of rapamycin (STAT3/mTOR) signaling pathway, and to decipher the possible mechanism. MethodSixty female SLE patients who met the criteria in the First Affiliated Hospital of Anhui University of Chinese Medicine from May 2022 to May 2023 were selected and randomized into a control group and an observation group (30 cases in each group). The control group was treated with prednisone acetate + hydroxychloroquine sulfate orally, and the observation group was additionally treated with Qihuang Jianpi Zishen granules. The treatment lasted for 8 weeks. The SLE disease activity (SLEDAI), TCM syndrome score, erythrocyte sedimentation rate (ESR), hypersensitive C-reactive protein (hs-CRP), immune indexes [immunoglobulin G (IgG), C3, C4, CD4+, and CD8+], interleukin (IL)-17, IL-23, interferon (IFN)-γ, 24 h urinary protein (24 h PRO), serum creatinine (SCr), and expression of proteins [STAT3, phosphorylated (p)-STAT3, mTOR protein and STAT3,mTOR mRNA] in the STAT3/mTOR signaling pathway were determined before and after treatment. In addition, the adverse reactions were recorded. ResultAfter 8 weeks of treatment, the total response rate in the observation group was 93.33% (28/30), which was higher than that (70.00%, 21/30) in the control group (χ2=4.007, P<0.05). After treatment, both groups showed declined SLEDAI, TCM syndrome score, ESR, hs-CRP, IgG, CD8+, IL-17, IL-23, IFN-γ, 24 h PRO, SCr, and expression of proteins in the STAT3/mTOR pathway (P<0.01) and elevated levels of C3, C4, and CD4+ (P<0.01). Moreover, the observation group had lower SLEDAI, TCM syndrome score, ESR, hs-CRP, IgG, CD8+, IL-17, IL-23, IFN-γ, 24 h PRO, SCr, and expression of proteins in the STAT3/mTOR pathway (P<0.05, P<0.01) and higher levels of C3, C4, and CD4+ (P<0.05, P<0.01) than the control group after treatment. Neither group showed serious adverse reactions during the treatment period. ConclusionQihuang Jianpi Zishen Granules can ameliorate the inflammatory response, reduce the disease activity, and mitigate the kidney injury in SLE by inhibiting the STAT3/mTOR signaling pathway to regulate the immune function.
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ObjectiveExploring the role of microRNA126 (miRNA126) in chronic kidney disease combined with atherosclerosis (CKD AS) by regulating the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway and the mechanism of Shenshuai Xiezhuo decoction in the intervention of CKD AS rats with 5/6 nephrectomy combined with high-fat feeding. MethodA total of 60 SD rats were randomly divided into sham operation group, model group, losartan group, and low, medium, and high dose groups of Shenshuai Xiezhuo decoction. The CKD AS rat model was established by 5/6 nephrectomy combined with high-fat feeding for 10 weeks. The low, medium, and high dose groups (6.0, 12.0, 24.0 g·kg-1·d-1) of Shenshuai Xiezhuo decoction and the losartan group (20 mg·kg-1·d-1) were gavaged, and the corresponding intervention was carried out for eight weeks. Then, the rats were killed, and samples were collected for corresponding detection. Fully automated biochemical analyzers were used to detect kidney function and blood lipids in rats: blood creatinine (SCr), blood urea nitrogen (BUN), total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) levels. Hematoxylin-eosin (HE) and Masson staining of aortic tissue and pathological observation under a light microscope were carried out, and autophagosomes and autophagy lysosomes were observed by transmission electron microscopy. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to determine the mRNA levels of miRNA126, PI3K, Akt, and mTOR in rats, and Western blot was used to determine the protein expression levels of phosphorylated (p)-PI3K, PI3K, p-Akt, Akt, p -mTOR, mTOR, benzyl chloride 1 (Beclin-1), and microtubule-associated protein light chain 3Ⅱ/Ⅰ (LC3Ⅱ/LC3Ⅰ). ResultCompared with the sham operation group, the serum SCr, BUN, TC, TG, and LDL-C in the model group were significantly increased (P<0.01). Compared with the model group, the SCr, BUN, TC, TG, and LDL-C were decreased in the losartan group and low, medium, and high dose groups of Shenshuai Xiezhuo decoction (P<0.05). Compared with the sham operation group, thickening plaques, infiltration of mononuclear macrophages, a small number of foam cells, disordered arrangement of smooth muscle fibers in the tunica media, and increased collagen fibers were observed in the model group, and the lesions in the losartan group and Shenshuai Xiezhuo decoction groups were alleviated compared with those in the model group. Compared with the model group, the number of autophagosomes and autophagy lysosomes increased in the medium and high dose groups of Shenshuai Xiezhuo decoction. Compared with the sham operation group, the expression of miRNA126 in the aortic tissue of the model group was significantly decreased (P<0.01), and the mRNA expressions of PI3K, Akt, and mTOR were significantly increased (P<0.01). Compared with the model group, the expression of miRNA126 in the aortic tissue of rats in high, medium, and low dose groups of Shenshuai Xiezhuo decoction and losartan group was significantly increased (P<0.01), while the mRNA expressions of PI3K, Akt, and mTOR were significantly decreased (P<0.01). Compared with the sham operation group, the protein expressions of p-PI3K, PI3K, p-Akt, Akt, p-mTOR, and mTOR in the model group were significantly increased (P<0.01), while the protein levels of Beclin-1, LC3Ⅰ, and LC3Ⅱ were significantly decreased (P<0.01). Compared with the model group, the protein expressions of p-PI3K, PI3K, p-Akt, Akt, p-mTOR, and mTOR in the losartan group and low, medium, and high dose groups of Shenshuai Xiezhuo decoction were decreased (P<0.05), while the protein levels of Beclin-1 and LC3Ⅱ/LC3Ⅰ were increased (P<0.05). ConclusionThe expression of miRNA126 is decreased in the aortic tissue of CKD AS rats, and the PI3K/Akt/mTOR pathway is activated to inhibit autophagy flux. Shenshuai Xiezhuo decoction regulates the PI3K/Akt/mTOR signaling pathway through miRNA126, restores the autophagy of aortic endothelial cells, protects the damage of CKD vessels, reduces the formation of As plaques, and slows the development of cardiovascular complications.
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AIM: To explore the intervention effect of Dahuangtang pellets (DHT) on diabetic nephropathy (DN) based on the AMP-activated protein kinase/mammalian target of rapamycin/unc-51-like kinase 1 (AMPK/mTOR/ULK1) signaling pathway. METHODS: Eight mice were randomly assigned to the model group, the dapagliflozin group, and the DHT (high, medium, and low dosage) group out of a total of 40 C57BL/KSJ-db/db (hereafter referred to as db/db) mice; another 10 C57BL/KSJ-db/dm mice were used as the normal group, saline was provided to the normal and model groups, and the mice in the treatment group received the appropriate medications. The medications were given for 10 consecutive weeks, once per day, to the mice in the treatment group. At weeks 0, 4, 8, and 10 of administration, fasting blood glucose (FBG) was assessed by drawing blood at a predetermined time from the tail vein; Urine samples were taken at 0, 5, and 10 weeks after treatment to evaluate the levels of albumin and creatinine, and the urinary albumin-creatinine ratio (ACR) was computed. After 10 weeks, mice in each group were assayed for 24 h total urine protein, serum creatinine (Scr), urea nitrogen (BUN) levels; Western blotting analysis was conducted to detect the expression of p-AMPK, p-mTOR, and p-ULK1, as well as the expression of autophagy related proteins homolog of yeast Atg6 (Beclin-1), autophagy-related proteins microtubule-associated protein 1 light chain 3 (LC3), P62 in renal tissue; Immunohistochemistry was used to measure the expression of podocyte lacunar membrane proteins (Nephrin, Podocin) in renal tissues; The pathological morphology of renal tissue was observed by light microscopy and transmission electron microscopy. RESULTS: Compared with the model group, FBG, ACR, and 24 h total urine protein were reduced in the dapagliflozin group and DHT groups of mice, and there was no statistically significant difference in Scr and BUN; In renal tissues, there is increased expression of p-AMPK and p-ULK1, decreased expression of p-mTOR, increased expression of LC3II / LC3I and Beclin-1, and decreased expression of P62 (P<0.01, P< 0.05); differentially upregulated in glomeruli are the podocyte lacunar membrane proteins Nephrin and Podocin (P<0.01, P<0.05); renal pathologic damage was reduced to varying degrees; transmission electron microscopy showed an increase in the number of autophagic vesicles and autophagic lysosomes. CONCLUSION: DHT can delay the development of DN by regulating the AMPK / mTOR / ULK1 signaling pathway, enhancing podocyte autophagy, and protecting glomeruli.
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Non-infectious chronic diseases in human including diabetes, non-alcoholic fatty liver disease (NAFLD), atherosclerosis (AS), neurodegenerative diseases, osteoporosis, as well as malignant tumors may have some common pathogenic mechanisms such as non-resolved inflammation (NRI), gut microbiota dysfunction, endoplasmic reticulum stress, mitochondria dysfunction, and abnormality of the mammalian target of rapamycin (mTOR) pathway. These pathogenic mechanisms could be the basis for "homotherapy for heteropathy" in clinic. Some commonly used clinical drugs, such as metformin, berberine, aspirin, statins, and rapamycin may execute therapeutic effect on their targeted diseases,and also have the effect of "homotherapy for heteropathy". The mechanisms of the above drugs may include anti-inflammation, modulation of gut microbiota, suppression of endoplasmic reticulum stress, improvement of mitochondria function, and inhibition of mTOR. For virus infectious diseases, as some viruses need certain commonly used replicases, the inhibitors of the replicases become examples of "homotherapy for heteropathy" for antiviral therapy in clinic (for example tenofovir for both AIDS and HBV infection). Especially, in case of outbreak of new emerging viruses, these viral enzyme inhibitors such as azvudine and sofibuvir, could be rapidly used in controlling viral epidemic or pandemic, based on the principle of "homotherapy for heteropathy". In this review article, we show the research progress of the biological basis for "homotherapy for heteropathy" and the possible mechanisms of some well-known drugs, in order to provide insights and new references for innovative drug R&D.
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ObjectiveTo explore the mechanism of Wenyang Jieyu prescription in regulating hippocampal neuron apoptosis and improving synaptic plasticity in the mouse model of depression induced by maternal separation combined with restraint stress. MethodThe mice on postnatal day 0 (PD0) were randomly assigned into a control group (n=10) and a modeling group (n=50). Maternal separation combined with restraint stress was adopted to establish the mouse model of depression, and the modeled mice were randomized into model, Wenyang prescription, Jieyu prescription, Wenyang Jieyu prescription, and fluoxetine groups (n=10) on the weaning day (PD21). From PD21 to PD111, the mice were fed with the diets mixed with corresponding medicines. The sucrose preference test, open field test, O-maze test, and novel object recognition test were then conducted to evaluate the depression, memory, and learning abilities of mice. Immunohistochemistry (IHC) was employed to measure the atomic absorbance (AA) of postsynaptic density protein 95 (PSD95) in the hippocampus. Terminal-deoxynucleoitidyl transferase-mediated nick-end labeling (TUNEL) was employed to detect the apoptosis of hippocampal neurons. Western blot was employed to determine the protein levels of brain-derived neurotrophic factor (BDNF), phosphorylated tyrosine kinase receptor B/tyrosine kinase receptor B (p-TrkB/TrkB), phosphorylated protein kinase B/protein kinase B (p-Akt/Akt), phosphorylated mammalian target of rapamycin/mammalian target of rapamycin (p-mTOR/mTOR), B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), cysteinyl aspartate-specific proteinase-3 (Caspase-3), synaptophysin (Syn), and PSD95. ResultCompared with the control group, the modeling decreased the sucrose preference rate, time spent in central zone within 5 min, total movement distance, time spent in the open arm, and cognition index (P<0.01). Furthermore, it decreased the expression of PSD95, increased the neuron apoptosis in the hippocampus (P<0.01), down-regulated the protein levels of BDNF, p-TrkB/TrkB, p-Akt/Akt, p-mTOR/mTOR, Bcl-2, PSD95, and Syn (P<0.01), and up-regulated the protein levels of Bax and Caspase-3 (P<0.05) in the hippocampus. Compared with the model group, Wenyang Jieyu prescription and fluoxetine increased the sucrose preference rate, time spent in central zone within 5 min, total movement distance, time spent in the open arm, and cognition index (P<0.05, P<0.01). Moreover, the drugs increased the expression of PSD95, reduced the neuron apoptosis (P<0.01), up-regulated the protein levels of BDNF, p-TrkB/TrkB, p-Akt/Akt, p-mTOR/mTOR, Bcl-2, PSD95, and Syn (P<0.01), and down-regulated the protein levels of Bax and Caspase-3 (P<0.01). ConclusionWenyang Jieyu prescription outperformed Wenyang prescription and Jieyu prescription in the treatment of the depressive behavior induced by maternal separation combined with restraint stress in mice. It exerted the therapeutic effect by reducing the hippocampal neuron apoptosis and improving the synaptic plasticity via the BDNF/Akt/mTOR pathway.