الملخص
Arbutin is utilized in traditional remedies to cure numerous syndromes because of its anti-microbial, antioxidant, and anti-inflammatory properties. This study aimed to evaluate chemopreventive effects of arbutin on azoxymethane (AOM)-induced colon aberrant crypt foci (ACF) in rats. Five groups of rats were used: normal control group (rats injected hypodermically with sterile phosphate-buffered saline once per week for two weeks) and groups 2-5, which were subcutaneously inoculated with 15 mg/kg AOM once a week for two weeks. AOM control and 5-fluorouracil (5-FU) control groups were fed 10% Tween orally daily for 8 weeks using a feeding tube. The treated groups were fed 30 and 60 mg/kg arbutin every day for 2 months. ACF from the AOM control group had aberrant nuclei in addition to multilayered cells and an absence of goblet cells. The negative control group displayed spherical cells and nuclei in basal positions. Histological examination revealed a reduced number of AFC cells from colon tissues of the 5-FU reference group. Arbutin-fed animals showed down-regulation of proliferating cell nuclear antigen (PCNA) and up-regulation of Bax protein compared to AOM control. Rats fed with arbutin displayed a significant increase of superoxide dismutase (SOD) and catalase (CAT) activities in colon tissue homogenates compared to the AOM control group. In conclusion, arbutin showed therapeutic effects against colorectal cancer, explained by its ability to significantly decrease ACF, down-regulate PCNA protein, and up-regulate Bax protein. In addition, arbutin significantly increased SOD and CAT, and decreased malondialdehyde (MDA) levels, which might be due to its anti-proliferative and antioxidant properties.
الملخص
Osteoarthritis (OA) is a highly prevalent joint disorder characterized by progressive degeneration of articular cartilage, subchondral bone remodeling, osteophyte formation, synovial inflammation, and meniscal damage. Although the etiology of OA is multifactorial, pro-inflammatory processes appear to play a key role in disease pathogenesis. Previous studies indicate that electroacupuncture (EA) exerts chondroprotective, anti-inflammatory, and analgesic effects in preclinical models of OA, but the mechanisms underlying these potential therapeutic benefits remain incompletely defined. This study aimed to investigate the effects of EA on OA development in a rat model, as well as to explore associated molecular mechanisms modulated by EA treatment. Forty rats were divided into OA, EA, antagomiR-214, and control groups. Following intra-articular injection of monosodium iodoacetate to induce OA, EA and antagomiR-214 groups received daily EA stimulation at acupoints around the knee joint for 21 days. Functional pain behaviors and chondrocyte apoptosis were assessed as outcome measures. The expression of microRNA-214 (miR-214) and its downstream targets involved in apoptosis and nociception, BAX and TRPV4, were examined. Results demonstrated that EA treatment upregulated miR-214 expression in OA knee cartilage. By suppressing pro-apoptotic BAX and pro-nociceptive TRPV4, this EA-induced miR-214 upregulation ameliorated articular pain and prevented chondrocyte apoptosis. These findings suggested that miR-214 plays a key role mediating EA's therapeutic effects in OA pathophysiology, and represents a promising OA treatment target for modulation by acupuncture.
الملخص
ObjectiveTo investigate the mechanism of modified Shenhong Tongluo prescription on cell apoptosis in rats with myocardial ischemia-reperfusion injury (MIRI). MethodSixty Sprague-Dawley (SD) rats were randomly divided into a blank group, a model group, low-, medium-, and high-dose groups of modified Shenhong Tongluo prescription, and a simvastatin group. Except for the blank group, a rat model of MIRI was prepared by ligating the left anterior descending coronary artery. Starting from the first day after successful modeling, the blank group (1.0 mL·kg-1 physiological saline), model group (1.0 mL·kg-1 physiological saline), low-, medium-, and high-dose groups of modified Shenhong Tongluo prescription (1.031, 2.063, and 4.126 g·kg-1 Shenhong Tongluo prescriptiona standard concentrate), and simvastatin group (0.71 mg·kg-1 simvastatin) were orally administered once daily for 2 weeks. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of cardiomyocytes. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum creatine kinase isoenzyme (CK-MB), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). TdT-mediated dUTP nick-end labeling(TUNEL) staining was used to detect the apoptosis rate of rat cardiomyocytes. Western blot was used to detect the expression levels of apoptosis-related proteins B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and caspase-3. ResultCompared with the blank group, in the model group, HE staining showed disturbed arrangement of cardiomyocytes, incomplete fibers, focal necrosis of cardiomyocytes, and inflammatory cell infiltration; serum CK-MB, IL-6, and TNF-α levels were significantly increased (P<0.05); apoptosis rate of cardiomyocytes was significantly increased (P<0.01), with significantly increased expression levels of Bax and Caspase-3 proteins, and significantly decreased Bcl-2 expression (P<0.05). Compared with the model group, the low-, medium-, and high-dose groups of modified Shenhong Tongluo prescription significantly reduced CK-MB, IL-6, and TNF-α levels (P<0.05), significantly downregulated cardiomyocyte apoptosis rate (P<0.05), significantly decreased Bax and Caspase-3 proteins, and significantly increased Bcl-2 expression levels (P<0.01). In the modified Shenhong Tongluo prescription groups, the expression levels of Bax and Caspase-3 proteins significantly decreased with increasing dosage, while the expression level of Bcl-2 significantly increased with increasing dosage of modified Shenhong Tongluo prescription (P<0.05). ConclusionShenhong Tongluo prescription can alleviate myocardial tissue pathological damage and reduce myocardial cell apoptosis, possibly by inhibiting Caspase-3 and Bax expression and promoting Bcl-2 expression.
الملخص
BACKGROUND:The clinical manifestation of acute radiation skin injury is recurrent necrotic ulcers,and its pathogenesis is still not fully understood.The establishment of a suitable animal model will have important clinical implications for the study of its pathogenesis,prevention and treatment. OBJECTIVE:To establish a model of acute β-ray radiation skin injury and to investigate the mechanism of injury. METHODS:Sixty-nine Sprague-Dawley rats were randomly divided into 30,45,60 Gy 32P-β-ray groups(n=21 per group)and control group(n=6).A single local irradiation of the back of the rats was performed using 32P radionuclide.The control group was operated in the same way as the irradiated groups except that it was not irradiated.The body mass and skin appearance of the rats were measured at 7,15,30,45,and 60 days after irradiation.Three rats from each group were selected at each observation time point.The skin injury was observed by hematoxylin-eosin staining,Masson staining,transmission electron microscopy,and TUNEL assay.P53,Bcl-2 and Bax protein levels in the skin were measured by immunohistochemistry and western blot assay. RESULTS AND CONCLUSION:There was no accidental death after irradiation,and the body mass of rats showed a gradual increase.The rats showed different degrees of epidermal necrosis,inflammatory cell infiltration,reduction of hair follicles and appendages,and collagen fibrillation,which were evident at 60 and 45 Gy.The levels of serum inflammatory factors,interleukin-6 and tumor necrosis factor-α,were significantly increased in a dose-dependent manner.Under the electron microscope,there are varying degrees of mitochondrial reduction,vacuolization and nuclear pyknosis in the cells.The degree of cell apoptosis showed a certain dose-dependence.Immunohistochemistry and western blot results showed an increase in the expression of P53 and Bax proteins and a decrease in the expression of Bcl-2 protein in the skin after irradiation.There were significant differences between the 60 Gy group and the 45 Gy and 30 Gy groups(P<0.05).To conclude,irradiation with 60 Gy and 45 Gy 32P radionuclide on the back of rats could successfully establish a practically pre-clinical animal model,and the mechanism is related to the up-regulation of P53 and Bax and the down-regulation of Bcl-2.This model can provide a reference for the establishment of animal models for the study of the mechanism of radiation skin injury and its prevention and treatment.
الملخص
BACKGROUND:The treatment of post-stroke dysphagia with Lipopharyngeal Qibi Formula has achieved good efficacy,and 5-hydroxytryptamine in peripheral serum and neurotransmitters in the nucleus tractus solitarius are closely related to swallowing.Therefore,this study was conducted to explore the modulatory effects of peripheral serum and nucleus tractus solitarius neurotransmitters in swallowing by using modern medical experimental methods such as molecular biology,thereby developing new ideas for the exploration of their mechanisms. OBJECTIVE:To verify the therapeutic effect of Lipopharyngeal Qibi Formula on post-stroke dysphagia and to investigate its mechanism of action. METHODS:Thirty-eight Sprague-Dawley rats were randomly divided into model group(n=14),treatment group(n=14)and sham-operated group(n=10).Animals in the model and treatment groups were modeled by reperfusion after 90 minutes of transient cerebral ischemia by wire bolus method.At 6 hours after modeling,neurological function was scored,and rats with a score of 2 were selected for subsequent experiments.The treatment group was given compound Lipopharyngeal Qibi Formula by gavage starting from the 2nd day after modeling and the remaining two groups were given normal saline by gavage.Changes in body mass,24-hour food and water intake were recorded on days 2,7,14 and 30.The swallowing initiation response time and the number of swallows were detected using a biosignal collector and a tonic transducer on days 14 and 30.After the swallowing test,the ischemic area of the brain in each group was measured by TTC staining.The expression of 5-hydroxytryptamine in the nucleus tractus solitarius of the medulla oblongata was measured by immunohistochemistry.The mRNA and protein expression levels of BCL-2 and BAX in the insula,premotor cortex,cingulate cortex and thalamus of rats in each group were measured by RT-PCR and Western blot,respectively. RESULTS AND CONCLUSION:Compared with the sham-operated group,the body mass,24-hour food intake and water intake were reduced,the swallow initiation response time was prolonged,and the number of swallows was reduced in the treatment and model groups at day 14 of gavage(P<0.05).Compared with the model group,the body mass,24-hour food intake and water intake of rats were increased in the treatment group at day 30 of gavage(P<0.05),but were still lower than those in the sham-operated group.Compared with the model group,the swallow initiation reaction time was shortened and the number of swallows increased in the treatment group,but the number of swallows was still significantly lower than that in the sham-operated group(P<0.05).Cerebral ischemia area was reduced in the treatment group compared with the model group,and the number of 5-hydroxytryptamine-positive cells in the nucleus tractus solitarius of the medulla oblongata was increased in the treatment group compared with the model group,but it was still significantly lower than that in the sham-operated group(P<0.05).Compared with the model group,the expression of BCL-2 mRNA and protein in the insula,cingulate cortex and thalamus of rats in the treatment group were significantly increased,the expression of BAX mRNA and protein were significantly decreased,and the BCL-2/BAX ratio was significantly increased(P<0.05).To conclude,the Chinese herbal compound Lipopharyngeal Qibi Formula could improve the number of swallows and swallowing initiation response time,as well as 24-hour food intake,body mass and other swallowing-related indexes in rats with post-stroke dysphagia.The mechanism of action may be achieved by improving the area of cerebral ischemia,inhibiting the apoptosis of neuronal cells in the insula,cingulate cortex and thalamus of rats,thus improving the regulation of the higher centers on the medulla oblongata swallowing center,and regulating the level of 5-hydroxytryptamine in the nucleus tractus solitarius.
الملخص
Objective @#To investigate whether norepinephrine (NE) regulates the oxidative stress in human endometrial epithelial cells (hEECs) by activating nuclear factor E2⁃related factor 2(Nrf2)/ heme oxygenase ⁃1(HO⁃1) signal pathway.@*Methods @#C ultured hEECs were used. The expression of α and β adrenergic receptors was detected by reverse transcription⁃polymerase chain reaction (RT⁃PCR) . Cell counting kit ⁃8(CCK⁃8) assay was applied to test the effect of NE on cell viability , then the cells were divided into C ontrol group and NE treatment group , and the appropriate concentrations were chosen. The expression of tight j unction proteins Occludin and zona occludens-1 (ZO⁃1) , apoptosis⁃related proteins apoptosis⁃related protein B ⁃cell lymphoma⁃2 protein(Bcl ⁃2) and Bcl ⁃2 associated X protein(Bax) , antioxidant proteins Nrf2 and HO⁃1 were examined by Western blot. The apoptosis was detected by flow cytometry. The malonaldehyde (MDA) and superoxide dismutase(SOD) in the cell culture medium were detected by enzyme⁃linked immunosorbent assays kit ( ELISA) .@*Results @#The mRNA expression of α1 a ,α1 b , α2 a , α2 b , α2 c , β1 , β3 was detected in the hEECs. After the NE treatment , no significant change in cell viability was ob served in low concentration (5 μmol/L and 10 μmol/L) groups , while 15 μmol/L and 20 μmol/L NE treatments for 6 h or 24 h promoted the cell viability significantly. The expression of ZO⁃1 and Occludin increased significantly in 15 μmol/L group after 24 h treatment , the expression of ZO⁃1 decreased in 6 h treatment group , significant down regulation was ob served after 15 μmol/L NE application , the expression of Occludin increased in 6 h group. The cell apoptosis increased compared with the control group after NE stimulation , espeserved after 24 h treatment. The ration of Bcl ⁃2/Bax > 1 . The expression of Nrf2 and HO⁃1 was elevated by NE. There was no obvious change in MDA level while significant elevation in SOD was detected in cell culture medium.@*Conclusion@#Nrf2/H0-1 signal is activated after application of NE to the hEECs, which may responsible for the upregulation of SOD, antioxidant and anti-apoptotic effect in the hEECs.
الملخص
ObjectiveTo investigate the mechanism of salvianolic acid F (Sal F) in repairing the high glucose-induced injury in human kidney-2 (HK-2) cells via the B-cell lymphoma-2 (Bcl-2)-associated X protein (Bax)/cysteinyl aspartate-specific proteinase 3 (Caspase-3)/gasdermin-E (GSDME) pathway. MethodThe cell counting kit-8 (CCK-8) was used to measure the relative viability of HK-2 cells exposed to high glucose and different concentrations (2.5, 5, 10, 20 μmol·L-1) of Sal F and the relative viability of HK-2 cells treated with Sal F for different time periods. The levels of lactate dehydrogenase (LDH) and interleukin-1β (IL-1β) in the supernatant of the cell culture were measured by the LDH assay kit and enzyme-linked immunosorbent assay (ELISA) kit, respectively. Flow cytometry combined with Annexin V-FITC/propidium iodide (PI) and Hoechst 33342/PI staining was employed to reveal the proportion of PI-positive HK-2 cells exposed to high glucose. Western blotting was employed to determine the protein levels of Bax, Bcl-2, cytochrome C, cysteinyl aspartate-specific proteinase (Caspase)-9, Caspase-3, and GSDME in the HK-2 cells exposed to high glucose and treated with Sal F. The 2,7-dichlorodihydrofluorescein diacetate fluorescence probe (DCFH-DA) and mitochondrial membrane potential assay kit (JC-1) were used to determine the production of reactive oxygen species (ROS) and the mitochondrial membrane potential in the HK-2 cells exposed to high glucose and treated with Sal F. ResultCompared with the blank group, the model group showed decreased cell viability (P<0.01), elevated levels LDH and IL-1β, increased proportion of PI-positive cells (P<0.01), up-regulated protein levels of Bax, cytochrome C, Caspase-9, Caspase-3, and GSDME (P<0.01), down-regulated protein level of Bcl-2 (P<0.01), decreased mitochondrial membrane potential, and excessive ROS accumulation. Compared with the model group, Sal F repaired the high glucose-induced injury in HK-2 cells (P<0.05), lowered the levels of LDH and IL-1β (P<0.05, P<0.01), and decreased the proportion of PI-positive cells (P<0.01). In addition, Sal F down-regulated the protein levels of Bax, cytochrome C, Caspase-9, Caspase-3, and GSDME and up-regulated the protein level of Bcl-2 (P<0.05, P<0.01), increased the mitochondrial membrane potential, and decreased the accumulation of ROS in HK-2 cells. ConclusionSal F can reduce the production of ROS, restore the balance of mitochondrial membrane potential, and inhibit pyroptosis via the Bax/Caspase-3/GSDME signaling pathway to repair the high glucose-induced injury in HK-2 cells.
الملخص
Aim To predict the mechanism of Fufang Congrong Yizhi Capsules (FCYC) in the treatment of mild cognitive impairment (MCI) by network pharmacology method, and further validate it in combination with cellular experiments. Methods TCMSP, Gene-Cards, OMIM and TTD databases, Chinese Pharmacopoeia and related literature were used to screen the active ingredients of FCYC and the targets of MCI treatment. The TCM-compound-target-disease network and PPI of intersection targets were constructed, and the GO and KEGG analysis were performed by the Ehamb bioinformation platform. GO and KEGG analysis were performed through Yihanbo biological information platform. Cell model of MCI was established by PC-12 injury induced by Aβ
الملخص
Introdução: O líquen plano oral é uma doença crônica imunologicamente mediada relativamente comum, que acomete a mucosa oral. Clinicamente, o LPO é classificado em seis padrões bem identificados: placa, reticular, bolhoso, atrófico, papular e erosivo.Sendo os mais comuns oos tipos reticulares e erosivos. A ativação dos linfócitos TCD4+ no LPO, pode induzir os ceratinócitos ao processo de apoptose através da respostaimunológica citotóxica. A proteína Bax desempenha uma função relevante para o processo apoptótico. Deste modo, a presente pesquisa consistiu em um estudo transversal retrospectivo, descritivo, quantitativo e comparativo. Objetivo: Avaliar a expressão imuno-histoquímica das proteínas MMP9 e Bax no LPO. Método: Foram utilizados 43 casos de LPO para análise da imunoexpressão de Bax e MMP-9. Os resultados foram analisados através dos testes estatísticos apropriados e serão considerados significativos, valores onde p<0,05. Resultado: A imunoexpressão de MMP9 foi significativamente maior nos ceratinócitos e quando analisados os subtipos de líquen plano oral, não foram observados diferenças estatísticas entre os tipos reticulares e erosivos para as proteínas analisadas. Conclusões: Com essas observações, infere-se que a alteração na expressão das proteínas estudadas sugere um distúrbio nos mecanismos apoptóticos, os quais estão associados às lesões de LPO, e podemos concluir também que as imunoexpressões dessas proteínas não apresentaram diferença, quando relacionada ao tipo clínico reticular ou erosivo. Com esse resultado pode-se contribuir para um maior entendimento sobre os possíveis mecanismos celulares envolvidos na etiopatogenia dessa lesão (AU).
Background: Oral lichen planus is a relatively common immune-mediated chronic disease that affects the oral mucosa. Clinically, OLP is classified into six well-identified patterns: plaque, reticular, bullous, atrophic, papular, and erosive. The most common being the reticular and erosive types. The activation of TCD4+ lymphocytes in the OLP can induce keratinocytes to the process of apoptosis through the cytotoxic immune response. Thus, the present research consisted of a retrospective, descriptive, quantitative and comparative crosssectional study. Objective: to evaluate the immunohistochemical expression of MMP-9 and Bax proteins in OLP. Methods: We used 20 cases of Inflammatory Fibrous Hyperplasia as control. The results were analyzed through the appropriate statistical tests and will be considered significant, values where p<0.05. Results: The immunoexpression of MMP-9 was significantly higher in keratinocytes and when the subtypes of oral lichen planus were analyzed, no statistical differences were observed between the reticular and erosive types for the proteins analyzed. Conclusions: With these observations, it is inferred that the alteration in the expression of the studied proteins suggests a disturbance in the proliferative and apoptotic mechanisms, which are associated with a pathological behavior of the oral mucosa, and consequently with a repercussion on the lesions of OLP, and we can also conclude that the immunoexpression of these proteins had no difference, when related to the reticular or erosive clinical type. This research aims to contribute to a greater understanding of the possible cellular mechanisms involved in the etiopathogenesis of this lesion, thus enabling the understanding of the clinical aspects of the pathology (AU).
الموضوعات
Lichen Planus, Oral/metabolism , Matrix Metalloproteinase 9/metabolism , bcl-2-Associated X Protein/metabolism , Medical Records , Epidemiology, Descriptive , Cross-Sectional Studies/methods , Retrospective Studies , Analysis of Variance , Data Interpretation, Statistical , Statistics, Nonparametric , Diagnosis, Differential , Mouth Mucosa/injuriesالملخص
ObjectiveTo study the effect of Bushen Huoxuetang on the apoptosis and the expression of B-cell lymphoma (Bcl-2)-associated X protein (Bax)/ Bcl-2 and cleaved cysteine-containing aspartate proteolytic enzyme-3 (cleaved Caspase-3) in the nude mouse model of bone metastasis of breast cancer, and explore the mechanism of Bushen Huoxuetang in inhibiting bone destruction. MethodThirty BALB/c female nude mice were randomly assigned into blank group (n=6) and model group (n=24). The suspension of 4T1 breast cancer cells was injected into the tibia of mouse right lower limb to establish model of bone metastasis of breast cancer. The successfully modeled nude mice were randomly assigned into model group, Bushen Huoxuetang group, zoledronic acid group, and combined drug group, with 6 mice in each group. Bushen Huoxuetang was administrated at a dose of 36.67 g·kg-1, once a day, and zoledronic acid was administrated by subcutaneous injection at a dose of 100 μg·kg-1, twice a week. The combined drug group was administrated with the same doses of Bushen Huoxuetang group by gavage and zoledronic acid by subcutaneous injection. The mice in the blank group and the model group were administrated with the same volume of distilled water by gavage for 14 days. On the next day at the end of drug administration, the mice were sacrificed by cervical dislocation. The general situation and weight changes of the mice were examined. The right lower limb was collected, and X-ray scanning and hematoxylin-eosin (HE) staining methods were used for observation of pathological changes in the bone. The terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) was employed to detect the apoptosis of bone tissue in nude mice, and Western blot to determine the expression of Bax/Bcl-2 and cleaved Caspase-3 in the bone tissue. ResultCompared with the blank group, the modeling reduced the body weight (P<0.01) and increased the right lower limb weight of the nude mice (P<0.01). Compared with the model group, Bushen Huoxuetang, zoledronic acid, and their combination increased the body weight (P<0.01) and decreased the right lower limb weight (P<0.01). Compared with the blank group, the other groups showed obvious tumor cell atypia, deep nuclear staining, and clear bone metastasis, and the model group showed obvious osteolytic damage in right lower limb and loss of proximal tibia and knee joint. Compared with the model group, Bushen Huoxuetang, zoledronic acid, and their combination reduced the osteolytic lesions in the right lower limb and recovered part of the bone structure, demonstrating an inhibitory effect on bone destruction. The TUNEL assay showed that the model group had lower apoptosis rate of bone metastatic tumor cells than the blank group, Bushen Huoxuetang group, zoledronic acid group, and combined drug group (P<0.01). Compared with the blank group, the modeling down-regulated the expression of Bax and cleaved Caspase-3 (P<0.01) and up-regulated the expression of Bcl-2 (P<0.01). Compared with the model group, Bushen Huoxuetang, zoledronic acid, and their combination up-regulated the expression of Bax (P<0.01) and cleaved Caspase-3 (P<0.05, P<0.01) and down-regulated the expression of Bcl-2 (P<0.05, P<0.01). ConclusionBushen Huoxuetang may inhibit bone destruction in the nude mouse model of bone metastasis of breast cancer by up-regulating the expression of Bax, down-regulating the expression of Bcl-2, activating cleaved Caspase-3, and further inducing apoptosis.
الملخص
Islet transplantation is a promising treatment of diabetes mellitus and its complications. Nevertheless, dysfunction post-transplantation, rejection and shortage of donors are the bottleneck issues in the field of islet transplantation. Optimizing the preservation method of pancreas plays a positive role in obtaining a sufficient quantity of effective islets and maintaining their functions. During the culture stage, anti-rejection and anti-apoptosis treatment of islets, including mesenchymal stem cell (MSC), MSC-derived exosomes, anti-apoptosis drugs and gene modification, may become major approaches for islet protection and functional maintenance in clinical islet transplantation. Use of anti-instant blood-mediated inflammatory reaction (IBMIR) drugs after islet transplantation also plays a critical role in protecting islet function. In this article, the whole process from islet preparation to islet transplantation was illustrated, and relevant strategies of islet protection and functional maintenance were reviewed, aiming to provide reference for improving the quality of donors to compensate for the shortage of absolute quantity of donors and elevating the efficiency of islet transplantation.
الملخص
ObjectiveTo investigate the protective effect of Geju Hugan tablets on the liver of mice with alcohol-induced liver injury, and explore the underlying mechanism based on nuclear factor-κB p65 (NF-κB p65) and B-cell lymphoma-2 (Bcl-2)/Bcl-2-associated X protein (Bax) signaling pathways. MethodAccording to the body weight, 60 SPF-grade male ICR mice were randomized into normal, model, Compound Yiganling tablets (0.16 g·kg-1), and low-, medium-, and high-dose (0.2, 0.4, 0.8 g·kg-1, respectively) Geju Hugan tablets groups. The drugs were administrated at the corresponding doses by gavage, and the normal and model groups with equal volume of pure water once a day for 28 consecutive days. On day 29, the mice in other groups except the normal group were administrated with liquor (53% Vol) by gavage twice a day at the doses of 20, 10 mL·kg-1 and with the interval of 6 h. Samples were harvested on day 30. The histopathological changes in the liver were observed by hematoxylin-eosin (HE) staining, and the ultrastructural changes in hepatocytes were observed by transmission electron microscopy. The enzyme-linked immunosorbent assay was employed to measure the levels of malonaldehyde (MDA), reduced glutathione (GSH), and triglycerides (TG) in the liver tissue and the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum. Western blotting was employed to determine the protein levels of NF-κB p65, phosphorylated p-inhibitor kappa B alpha (p-IκBα), Bcl-2, and Bax in the liver tissue. ResultCompared with the normal group, the model group showed increases in the ALT, AST, MDA, and TG levels, a decrease in the GSH level, and increases in the liver injury scores evaluated based on the HE, oil red O, and transmission electron microscopy (P<0.01). Moreover, the model group showed up-regulated expression of NF-κB, p-IκBα, and Bax (P<0.05, P<0.01) and down-regulated expression of Bcl-2 (P<0.05) in the liver tissue. Compared with the model group, Geju Hugan tablets of all the doses lowered the ALT, AST, MDA, and TG levels and elevated the GSH level (P<0.01). The liver injury scores assessed based on HE staining and transmission electron microscopy in the medium- and high-dose Geju Hugan tablets groups were lower than those in the model group (P<0.01). Compared with the model group, medium- and high-dose Geju Hugan tablets down-regulated the protein levels of NF-κB, p-IκBα, and Bax (P<0.01) and all doses of Geju Hugan tablets up-regulated the protein level of Bcl-2 (P<0.01). ConclusionGeju Hugan tablets protect mice from alcohol-induced liver injury by down-regulating NF-κB signaling pathway to alleviate inflammation in the liver tissue and down-regulating the expression of Bax and up-regulating the expression of Bcl-2 to inhibit hepatocyte apoptosis.
الملخص
ObjectiveTo investigate the mechanism of ethyl acetate extract of Tibetan medicine dampness bud Gentianopsis paludosa in the prevention and treatment of recurrent ulcerative colitis (UC) in rats with dampness-heat in large intestine syndrome based on the apoptotic pathway mediated by the B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax). MethodUsing the disease-syndrome combination method, a recurrent UC model of dampness-heat in large intestine syndrome was constructed in rats. Seventy SPF-grade male SD rats were randomly divided into control group, model group, high-, medium-, and low-dose ethyl acetate of G.paludosa groups (150, 75, 37.5 mg·kg-1), and mesalazine group (135 mg·kg-1). The rats were orally administered with respective drugs for 14 days. The general conditions of the rats were recorded, and colon length and mucosal damage were observed. The colon wet weight index and organ coefficients of the liver, spleen, and thymus were calculated. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of interleukin-6 (IL-6) and interleukin-1β (IL-1β) in the serum of each group. Hematoxylin-eosin (HE) staining was performed to observe pathological changes in the colon. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was used to detect apoptosis in colonic epithelial cells. Western blot was used to measure the expression levels of Bcl-2, Bax, Caspase-3, Caspase-9, Zona Occludens-1 (ZO-1), Claudin3, and Occludin in colonic tissue. Immunohistochemistry (IHC) was used to observe the expression of Bax and Caspase-3 in colonic epithelial cells. ResultCompared with the control group, the model group showed significant increases in the disease activity index (DAI) score, colonic mucosal damage index (CMDI), intestinal epithelial apoptosis, liver and spleen indexes, and levels of inflammatory factors IL-1β and IL-6 in the serum (P<0.01), decreased expression of intestinal mucosal protective proteins ZO-1, Claudin3, and Occluding (P<0.01), increased expression of pro-apoptotic proteins Bax, Caspase-3, and Caspase-9 (P<0.01), and decreased expression of anti-apoptotic protein Bcl-2 (P<0.01). Compared with the model group, the high-, medium-, and low-dose ethyl acetate of G.paludosa groups all significantly improved the general condition of the rats, reduced colonic lesions, decreased intestinal epithelial cell apoptosis, reduced liver and spleen indexes, upregulated the expression of ZO-1, Claudin3, Occludin, and Bcl-2 proteins, and downregulated the expression of Bax, Caspase-3, and Caspase-9 proteins, with the high- and medium-dose ethyl acetate of G.paludosa groups showing the superior effects (P<0.05, P<0.01). ConclusionEthyl acetate of G.paludosa can alleviate colonic mucosal damage and exert a therapeutic effect on UC by regulating the Bcl-2/Bax signaling pathway.
الملخص
Objective To investigate the effect of the SMAC gene on paclitaxel sensitivity and cellular activity in lung adenocarcinoma cells based on the caspase-3/Bcl-2/Bax signaling pathway. Methods A paclitaxel-resistant cell line A549/Taxol was established for lung adenocarcinoma, and the cells were divided into four following groups: pcDNA-NC (transfected with pcDNA-NC blank vector), pcDNA-SMAC (transfected with pcDNA-SMAC vector), siRNA-NC (transfected with siRNA-NC empty virus vector), and siRNA-SMAC groups (transfected with siRNA-SMAC lentiviral vector). The SMAC mRNA expression in cells was detected by qRT-PCR; cell sensitivity was detected by MTT; cell proliferation ability was detected by cloning assay; cell invasion ability was detected by Transwell; apoptosis ability was detected by flow cytometry assay; and caspase-3, Bcl-2 and Bax protein expression in cells were detected by Western blot analysis. Results The SMAC mRNA expression was significantly lower in A549 cells compared with BEAS-2B cells (P < 0.05). The SMAC mRNA expression was significantly higher in the pcDNA-SMAC group than that in the pcDNA-NC group cells (P < 0.05). The SMAC mRNA expression was significantly lower in the cells of the siRNA-SMAC group (P < 0.05) than that in the siRNA-NC group. The SMAC mRNA expression was significantly lower in the cells of the siRNA-SMAC group (P < 0.05) than in the siRNA-NC group. Compared with the pcDNA-NC group, the cell IC50, cell clone number, cell invasion ability, and Bcl-2 protein and Bcl-2/Bax ratio were significantly lower in the pcDNA-SMAC group, the cell resistance index reversal was 2.51-fold, and the apoptosis ability and caspase-3, as well as Bax protein expression, were significantly higher (P < 0.05). Compared with the siRNA-NC group, cell IC50, cell clone number, cell invasion ability, and Bcl-2 protein and Bcl-2/Bax ratio were significantly higher in the siRNA-SMAC group, and apoptosis ability and caspase-3 and Bax protein expression were significantly lower (P < 0.05). Conclusion High expression of SMAC increases paclitaxel sensitivity, inhibits cell growth and invasion, promotes apoptosis in lung adenocarcinoma cells, and has a regulatory effect on the caspase-3/Bcl-2/Bax signaling pathway.
الملخص
Objective:To investigate the role and mechanism of N 6-methyladenosine (m 6A) methyltransferase-like 3 (METTL3) in vascular calcification (VC) of chronic kidney disease (CKD) through apoptosis-associated protein. Methods:(1) Real-time fluorescence quantitative PCR was used to test METTL3 mRNA in serum of maintenance hemodialysis (MHD) patients. (2) Western blotting was used to detect the expression of METTL3 protein in high-phosphorus stimulated vascular smooth muscle cells (VSMCs), and immunofluorescence double lable was used to observe the distribution of METTL3 and Runt-related transcription factor 2 (Runx2). The METTL3 overexpressed and knockdown plasmids were constructed and transfected into VSMCs. Alizarin red staining was used to detect calcification degree. Western blotting was used to detect the expressions of osteogenic markers [Runx2, bone morphogenetic protein-2(BMP-2), collagen Ⅰ] and apoptosis- related proteins Bax and Bcl-2. (3) SD rats were randomly divided into control group, CKD-VC group and S-adenosylhomocysteine (SAH) intervention group. The calcification of thoracic aorta was evaluated by von Kossa staining, and the protein expressions of Runx2, Bax and Bcl-2 were detected by immunohistochemistry and Western blotting.Results:(1) METTL3 mRNA expression in MHD patients with VC was significantly lower than that in non-VC patients ( P<0.05), and was negatively correlated with coronary artery calcium score ( r=-0.65, P<0.001). (2) The expression of METTL3 in VSMCs stimulated by high phosphorus was decreased and showed a time dependence. Immunofluorescence double label showed that METTL3 and Runx2 were co-expressed in the nucleus. METTL3 was overexpressed in high-phosphorus induced VSMCs, and the expressions of Runx2, collagen I and BMP-2 were significantly decreased, accompanied by the decrease of calcified nodules and Bax/Bcl-2 ratio (all P<0.05). Conversely, METTL3 knockdown aggravated VSMCs calcification by inducing apoptosis. (3) Furthermore, METTL3 inhibitor SAH was administered in vivo, and it was found that inhibition of METTL3 expression significantly increased the calcification of rat thoracic aorta, and the Bax/Bcl-2 ratio and Runx2 expression were up-regulated. Conclusions:Serum METTL3 level is reduced in MHD patients with VC. In vivo and in vitro studies demonstrate that METTL3 inhibits VC in CKD by mediating the apoptosis-related protein Bax/Bcl-2.
الملخص
Aim To investigate the mechanism of action of Shen-Fu decoction in the prevention and treatment of cardiogenic shock based on network pharmacology and animal experiments. Methods The relevant targets and signaling pathways of cardiogenic shock of Shen-Fu decoction were predicted by network pharmacology, and a cardiogenic shock rat model was created by coronary artery ligation. Before modeling, rats were given the appropriate dose of Shen-Fu decoction or saline by gavage for 14 days according to the group, and real-time mean arterial pressure (MAP) changes were recorded after successful modeling. HE method was used to detect the myocardial histopathological changes of cardiogenic shock. TUNEL method was employed to detect rat myocardial cell apoptosis, and Western blotting was applied to determine the expression levels of rat myocardial Bax, Bcl-2, caspase-3, cleaved caspase-3 proteins. Results A total of 51 potential active ingredients of Shen-Fu decoction were screened out by network pharmacology, 80 targets of co-action with cardiogenic shock, and 43 core targets of close relationship between proteins, and GO enrichment analysis revealed that the core proteins were involved in the biology process (BP), mainly involving positive regulation of apoptotic process. KEGG enrichment analysis showed signaling pathways involving atherosclerosis-related, apoptosis and other signaling pathways. The results of animal model validation showed that Shen-Fu decoction could increase the shock blood pressure of rats with cardiogenic shock and alleviate the pathological changes of myocardial tissue, reduce the degree of apoptosis of rat cardiomyocytes, reduce the expression level of caspase-3, cleaved caspase-3 and Bax protein in rat myocardial tissue, and improve the expression level of Bcl-2 protein in myocardial tissue of rats. Conclusions The potential active ingredient of Shen-Fu decoction may play a role in the prevention and control of cardiogenic shock rats by acting on the target Bax, Bcl-2 to regulate the apoptosis signaling pathway of cardiomyocytes.
الملخص
Aim To study the protective effect of trigonelline on H
الملخص
Aim To identify the molecular target of gabapentin in the treatment of postherpetic neuralgia(PHN). Methods The molecular target of gabapentin for PHN was analyzed by network pharmacology and molecular docking and confirmed by coprecipitation test. Rats were randomly divided into control group, model group, model+50 mg·kg-1 gabapentin group, model+100 mg·kg-1 gabapentin group, and model+200 mg·kg-1 gabapentin group, with nine rats in each group. The pain-related behaviors of the rats were measured at different time points. The mRNA and protein expressions of CACNA2D1, Bax, and Bcl-2 in rat spinal cord were determined by immunofluorescence, Western blot, and qPCR. Results CACNA2D1 was the target gene of gabapentin that determined via network pharmacology, molecular docking, and co-precipitation tests. After modeling, mechanical pain threshold and thermal pain threshold significantly decreased, and the number of apoptotic GABA cells significantly increased. However, after intraperitoneal injection of 50, 100, and 200 mg·kg-1 gabapentin, mechanical pain threshold and thermal pain threshold significantly increased(P<0.05), and the number of apoptotic GABA cells significantly decreased(P<0.01). Immunofluorescence and Western blot results showed that compared with the model group, with the increase of gabapentin concentration, the positive expression rate of Bax significantly decreased, and the positive expression rate of Bcl-2 and CACNA2D1 significantly increased. The mRNA expression levels of Bax, Bcl-2 and CACNA2D1 detected by qPCR were consistent with the results of immunofluorescence and Western blot. Conclusions Gabapentin up-regulates the expression of target protein CACNA2D1, inhibits the proapoptotic protein Bax, and promotes the expression of apoptotic inhibitor Bcl-2.
الملخص
Resumen El cáncer representa un problema de salud pública a nivel mundial, con altas tasas de incidencia y mortalidad en países desarrollados y no desarrollados. En la actualidad se están evaluando alternativas terapéuticas de origen natural, con el propósito de establecer tratamientos más eficientes y menos invasivos. Dado que la apoptosis es el tipo de muerte programada que experimentan las células cancerosas por los tratamientos con los fármacos antineoplásicos,el objetivo de esta investigación, fue evaluar in vitro la capacidad pro-apoptótica y citotóxica de los extractos de valeriana, sobre una línea celular de cáncer de mama (MCF-7). En este estudio las células MCF7 se cultivaron y trataron con diferentes concentraciones de los extractos de la raíz, hojas y tallos de Valeriana rígida y Valeriana decussata. La viabilidad celular se evaluó mediante el ensayo MTT. Para la determinación de la expresión génica de las proteínas anti y pro-apoptóticas (Bax, Bcl-2 y p53), se usó el ensayo de la PCR cuantitativa de transcripción inversa. Las diferentes concentraciones de los extractos (10-8 a 10-1 mg/mL) disminuyeron la viabilidad (proliferación) celular en concentraciones dependientes. Estos extractos indujeron la expresión génica de las proteínas Bax y Bcl-2, pero no de p53. La expresión de Bax fue mayor que la de Bcl-2 e indujo un elevado índice Bax/Bcl-2 (condición proapoptotica). En conclusión, se determinó que los extractos de Valeriana decussata y Valeriana rígida poseen efecto reductor de la viabilidad (proliferación) de la línea celular de cáncer de mama MCF-7, probablemente mediado por la alteración de la relación de las proteínas Bax y Bcl-2 vinculadas a la apoptosis.
Abstract Cancer represents a worldwide public health problem, with high incidence and mortality rates in developed and undeveloped countries. Currently, therapeutic alternatives of natural origin are being evaluated with the purpose of establishing more efficient and less invasive treatments. Apoptosis is the type of programmed death cancer cells undergo during treatment with anti-neoplastic drugs. Therefore, the aim of this research was to evaluate in vitro the pro-apoptotic and cytotoxic capacity of valerian extracts on a breast cancer cell line (MCF-7). In this study, MCF7 cells were cultured and treated with different concentrations of the extracts of the root, leaves and stems of Valeriana rígida and Valeriana decussata. Cell viability was assessed by the MTT assay. Quantitative reverse transcription PCR assays were used for the determination of gene expression of anti- and proapoptotic proteins (Bax, Bcl-2, p53). Different concentrations of the extracts (10-8 to 10-1 mg/mL) decreased cell viability (proliferation) in a concentration-dependent manner. These extracts induced gene expression of Bax and Bcl-2 proteins but not of p53. The expression of Bax was higher than that of Bcl-2, causing an elevated Bax/Bcl-2 ratio (proapoptotic condition). In conclusion, it was determined that Valeriana decussata and Valeriana rígida extracts have a viability (proliferation) reducing effect on the MCF-7 breast cancer cell line, probably mediated by altering the ratio of Bax and Bcl-2 proteins linked to apoptosis.
الملخص
OBJECTIVE@#To explore the effect of hypoxia on the chemosensitivity of B-acute lymphoblastic leukemia (B-ALL) cells to Vincristine (VCR) and the mechanisms.@*METHODS@#B-ALL cells SUP-B15, Nalm-6 and RS4;11 were selected as the research objects. The cells were divided into the control group and the hypoxia mimic group (CoCl2 pretreatment). The two groups were treated with VCR at different concentrations for 24 hours, CCK-8 was used to detect cell viability, flow cytometry was used to detect cell apoptosis, and Western bolt method was used to detect hypoxia inducible factor (HIF-1α), BAX, Bcl-2 and β-actin protein expression. Quantitative real-time fluorescent PCR (qRT-PCR) was used to detect BAX and β-actin mRNA levels.@*RESULTS@#CoCl2 could simulate hypoxic environment to induce the expression of HIF-1α. The cells SUP-B15 and RS4;11 of the hypoxia mimic group were lower sensitivity to VCR as compared with the control group; the apoptosis rate of the hypoxia mimic group was lower than that of the control group after 80 nmol/L VCR treatment. The expression levels of BAX protein and mRNA in the hypoxia mimic group were lower than those of the control group, and there was no significant difference in the expression levels of Bcl-2 protein between two groups.@*CONCLUSION@#Under hypoxic conditions, HIF-1α may mediate VCR resistance in B-ALL cells by downregulating the pro-apoptotic protein BAX.