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ABSTRACT Objective: To investigate nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH) and hepatic fibrosis in biopsies of people with obesity who underwent bariatric surgery and examine the possible association of different variables with a diagnosis of NAFLD and NASH. Materials and methods: Epidemiological, clinical and laboratory data from 574 individuals with obesity of both genders seen by the same physician between 2003 and 2009 who had a liver biopsy during bariatric surgery were examined. Results: Of the 437 patients included, 39.8% had some degree of liver fibrosis, 95% had a histologic diagnosis of NAFLD, and the risk factors were age ≥ 28 years and Homeostatic Model Assessment (HOMA) ≥ 2.5 (p = 0.001 and p = 0.016, respectively). In the NAFLD group, NASH was present in 26% of patients and the associated factors were aspartate aminotransferase and alanine aminotransferase index (AST/ALT) > 1, high-density lipoprotein cholesterol (HDL-c) < 40 mg/dL, total cholesterol (TC) ≥ 200 mg/dL, gamma-glutamyl transferase (GGT) > 38 U/L and triglycerides (TG) levels > 150 mg/dL. The independent risk factors were low HDL-c, elevated AST/ALT and high TG. Conclusion: The variables associated with a diagnosis of NAFLD were HOMA ≥ 2.5 and age ≥ 28 years. NASH was associated with low HDL-c, high TG and AST/ALT ≤ 1.
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Hepatic fibrosis （HF） is a pathological process of abnormal repair of liver tissue structure caused by chronic liver injury， and its pathogenesis has not been fully clarified. Related studies have shown that programmed cell death may be associated with the onset of HF， and traditional Chinese medicine （TCM） has a significant effect in regulating programmed cell death to intervene against HF. This article reviews the main mechanism of the influence of programmed cell death on HF and discusses the possible mechanism of TCM regulation of programmed cell death in improving HF， which provides new ideas for TCM prevention and treatment of HF.
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ObjectiveTo investigate the effect of phytoestrogen biochanin A （BCA） on liver fibrosis induced by CCl4 in female mice with bilateral oophorectomy （ovariectomized） and its mechanism. MethodsA total of 50 ovariectomized Kunming mice were selected and given intraperitoneal injection of CCl4 to establish a model of liver fibrosis， and then according to body weight， they were randomly divided into model group， positive control group， and low-， middle-， and high-dose BCA groups， with 10 mice in each group. In addition， 10 female mice in the same litter were given resection of a small amount of adipose tissue near both ovaries to establish the sham-operation group. The mice in the positive control group were given estradiol 2 mg/kg by gavage， and those in the low-， middle-， and high-dose BCA groups were given BCA by gavage at a dose of 25， 50， and 100 mg/kg， respectively， once a day for 7 consecutive weeks； the mice in the sham-operation group and the model group were given an equal volume of 0.5% sodium carboxymethyl cellulose solution by gavage. The mice were anesthetized and sacrificed after administration to collect samples. Liver index and uterus index were measured； HE staining and Masson staining were used to observe liver histopathological changes； the biochemical analysis was used to measure the activity of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）； ELISA was used to measure the levels of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in liver tissue， and Western blot was used to measure the relative protein expression levels of collagen Ⅰ， transforming growth factor-beta 1 （TGF-β1）， alpha-smooth muscle actin （α-SMA）， estrogen receptor beta （ERβ）， and p-NF-κBp65/NF-κBp65 in liver tissue. The t-test was used for comparison of continuous data between two groups； a one-way analysis of various was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups and further comparison between two groups. ResultsCompared with the sham-operated group， the model group had a significant increase in liver index and a significant reduction in uterus index， as well as significant increases in the activities of serum AST and ALT， the levels of IL-6 and TNF-α in liver tissue， and the protein expression levels of collagen Ⅰ， TGF-β1， α-SMA， and p-NF-κBp65/NF-κBp65 in liver tissue （all P<0.05）， with no significant change in the expression of ERβ in liver tissue （P>0.05）， and the model group showed significant fibrosis lesions in the liver， such as hepatocyte edema， steatosis， and necrosis with inflammatory cell infiltration and hyperplasia， deposition， and staggered distribution of collagen fibers. Compared with the model group， the low-， middle-， and high-dose BCA groups had significant reductions in liver index， the activities of serum AST and ALT， the levels of IL-6 and TNF-α， and the protein expression levels of collagen Ⅰ， TGF-β1， α-SMA， and p-NF-κBp65/NF-κBp65 in liver tissue （all P<0.05）， with no significant change in uterine index （P>0.05）， as well as a significant increase in the protein expression level of ERβ in liver tissue （P<0.05） and varying degrees of improvement in liver fibrosis lesions. ConclusionBCA can effectively improve CCL4-induced liver fibrosis in ovariectomized female mice， possibly by upregulating ERβ to inhibit the NF-κB signaling pathway and then alleviating inflammatory response.
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ObjectiveTo quantitatively investigate the changes in the total volume and contour density of hepatic oval cells （HOC） in hepatic lobules of rats with carbon tetrachloride （CCl4）-induced hepatic fibrosis. MethodsA total of 11 healthy male Sprague-Dawley rats were randomly divided into control group with 5 rats and hepatic fibrosis group with 6 rats， and CCl4 and olive oil suspension were injected subcutaneously twice a week， 3 mL/kg each time. After five weeks of hepatic fibrosis modeling， five liver tissue blocks with a size of about 1 mm3 were randomly selected from the liver of each rat to prepare one Epon812 epoxy resin-embedded ultrathin section， and the stereological method and transmission electron microscopy were used for the quantitative analysis of the total volume and contour density of HOC in the hepatic lobules of rats. In addition， four liver tissue blocks with a thickness of 2 mm were randomly selected from the remaining liver of each rat to prepare two paraffin-embedded Masson staining sections， and the degree of liver fibrosis in each rat was qualitatively evaluated according to the Metavir staging criteria for liver fibrosis. The independent-samples t test was used for comparison of continuous data between groups. ResultsThe quantitative stereological analysis showed that the total volume of HOC in hepatic lobules was 15.40±7.63 mm3 in the control group and 146.80±114.00 mm3 in the liver fibrosis group， and compared with the control group， the total volume of HOC in hepatic lobules of rats in the liver fibrosis group was significantly increased by 8.53 times （t=-2.551， P=0.031）； the contour density of HOC in hepatic lobules was 56.20±40.40 in the control group and 566.50±317.00 in the liver fibrosis group， and compared with the control group， the contour density of HOC in hepatic lobules of rats in the liver fibrosis group was significantly increased by 9.08 times （t=-3.539， P=0.006）. Qualitative observation showed that liver fibrosis stage of rats reached stage Ⅱ-Ⅲ according to the Metavir scoring criteria， and massive proliferation of HOC was observed around the proliferation site of hepatic stellate cells in the perisinusoidal space of rats. ConclusionCCl4 induces significant proliferation of HOC in hepatic lobules of rats with liver fibrosis.
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ObjectiveTo investigate the preventive and therapeutic effects of Tiaogan Huaxian pills combined with entecavir on hepatic fibrosis in chronic hepatitis B with liver Qi stagnation， spleen deficiency， and blood stasis syndrome and its effect on diffusion-weighted imaging （DWI）. MethodClinical data of 117 patients with liver disease who visited the Department of Hepatology at the First Affiliated Hospital of Guangxi University of Chinese Medicine from January 2021 to April 2022 were retrospectively analyzed. According to different treatment plans， they were divided into a control group （59 cases） and a treatment group （58 cases）. Both groups of patients received entecavir-based etiology treatment， and the treatment group added Tiaogan Huaxian pills on the basis of basic treatment. Both groups were treated for 24 weeks. Before and after treatment， the two groups were compared in terms of alanine aminotransferase （ALT）， advanced surgical technologies （AST）， total bilirubin （TBil）， hepatitis B virus （HBV）-DNA conversion rate， liver stiffness measurement （LSM）， four items of liver fibrosis （hyaluronidase， type Ⅲ pro-collagen， type Ⅳ collagen， and laminin）， the fibrosis index based on four factors （FIB-4）， the aspartate aminotransferase to platelet ratio index （APRI）， the apparent diffusion coefficient （ADC） value in magnetic resonance imaging （MRI）， and traditional Chinese medicine symptom scores， so as to analyze the efficacy of the two groups. ResultBefore treatment， there was no significant difference in ALT， AST， TBil， LSM， four items of liver fibrosis， FIB-4， APRI， HBV-DNA conversion rate， ADC value， and traditional Chinese medicine symptom scores between the two groups. After treatment， both groups of patients showed significant reductions in ALT， AST， TBil， LSM， hyaluronidase， type Ⅲ pro-collagen， type Ⅳ collagen， laminin， FIB-4， and APRI （P<0.05） and a significant increase in ADC value （P<0.05） and HBV-DNA conversion rate （P<0.01）. The traditional Chinese medicine symptom score of the treatment group decreased significantly （P<0.05）. Compared with the control group after treatment， the effective rate of clinical traditional Chinese medicine in the treatment group was 91.38% （53/58）， which was significantly higher than that of the control group （54.23%， 32/59） （Z=-4.325， P<0.01）. In the treatment group， ALT， AST， TBil， LSM， hyaluronidase， type Ⅲ pro-collagen， type Ⅳ collagen， laminin， FIB-4， APRI， and traditional Chinese medicine symptom scores all decreased significantly （P<0.05）， and the increase in ADC values was more significant （P<0.05）， while the difference in HBV-DNA conversion rate was not statistically significant. There were no serious adverse reactions or events in either group. ConclusionTiaogan Huaxian pills combined with entecavir have significant clinical efficacy in the treatment of hepatic fibrosis in chronic hepatitis B， which can reduce liver inflammation activity， delay hepatic fibrosis progression， and reduce traditional Chinese medicine symptom scores. It is worthy of clinical promotion and application.
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Background: Previous studies have shown that hepatic fibrosis indices and rates can be used to predict cardiovascular mortality and morbidity. Our aim with this study was to investigate the effect of aspartate aminotransferase/alanine aminotransferase (AST/ALT) ratio and fibrosis?4 (FIB?4) index calculated with ALT, AST, and platelet biomarkers, which are simple, fast, and relatively inexpensive and were used in previous studies to predict cardiovascular disease prognosis, on the prediction of postoperative morbidity and early mortality after mitral valve replacement (MVR) surgery. Methods: By scanning the hospital electronic health record system, 116 patients who underwent isolated MVR or MVR + tricuspid valve intervention were identified from 178 patients who underwent MVR with the standard sternotomy procedure between 2011 and 2021. The study was completed with 81 of these patients. Patients were divided into AST/ALT <2 (Group 1) and >2 (Group 2). In addition, the same patients were divided into FIB?4 index <3.25 (Group 3) and >3.25 (Group 4), and a total of four groups were formed. Results: The mean age of Group 2 was significantly higher than Group 1 (P = 0.049). In addition, the mean age of Group 4 was significantly higher than Group 3 (P = 0.003). Postoperative complications did not differ between Groups 1 and 2 (P > 0.05). While noninvasive mechanincal ventilation (NIMV) requirements did not differ between Groups 3 and 4 (P > 0.05), MV duration and intensive care unit stay were significantly longer in Group 4 (P < 0.05). Conclusion: The AST/ALT ratio, which has been shown to be a predictor of cardiovascular mortality in various studies, was not useful in predicting mortality and morbidity in our study. However, a high FIB?4 index, another hepatic fibrosis index, was found to be associated with increased perioperative bleeding, duration of mechanical ventilation, and cardiac intensive care unit stay, which are important criteria in the prediction of morbidity in cardiovascular surgery.
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ObjectiveTo investigate the possible mechanism of compound Yinqi Sanhuang Jiedu Decoction （茵芪三黄解毒汤， YSJD） against the progression of hepatic fibrosis （HF）. MethodsThirty-eight C57BL/6J mice were randomly divided into blank group， model group， silybin group and low-， medium- and high-dose YSJD groups， with eight mice in the model group and six mice each in other groups. Except for the blank group， all mice were injected intraperitoneally with 10% carbon tetrachloride （CCl4） to establish a HF model， twice a week for 8 weeks. The drug intervention started one week after the first modeling； the low-， medium- and high-dose YSJD groups were given 8.325， 16.65 and 33.3 g/（kg·d） of YSJD suspension by gavage， respectively， while the silybin group was given 55 mg/（kg·d） of silybin suspension by gavage， and the blank group and the model group were given 0.2 ml normal saline by gavage， all for 8 weeks. The liver hardness of living mice was observed using a small animal ultrasound detector， and grey-scale ultrasound was recorded. The liver tissue was observed by Sirius scarlet staining， and the proportion of collagen fiber deposition was calculated. Liver function indicators including alanine aminotransferase （ALT） and aspartate aminotransferase （AST）， and serum laminin （LN）， hyaluronic acid （HA）， pre-collagen type III （PCIII） and collagen type IV （CIV） were also detected. The protein expression of transforming growth factor β1 （TGF-β1）， Vimentin， E-cadherin， α smooth muscle actin （α-SMA） in liver tissue were detected. ResultsCompared to the blank group， the model group showed increased gray value， collagen deposition，serum ALT， AST， HA， LN and PCIII levels， decreased expression of E-cadherin， and increased expression of N-cadherin，α-SMA，Vimentin and TGF-β1 in liver tissues （P＜0.05）. Compared to the model group， the ultrasonic gray value and the proportion of collagen fiber deposition in liver of silybin group and YSJD medium- and high-dose groups decreased， and the serum ALT， AST， LN， HA and PCⅢ levels decreased. Compared to the model group， the expression of E-cadherin in liver tissues of silybin group and all three YSJD groups increased， while the expressions of N-cadherin， Vimentin， TGF-β1 and α-SMA decreased （P＜0.05）， and among them， most improvements were seen in the medium-dose YSJD group （P＜0.05）. ConclusionThe effect of YSJD on significantly reducing the extent of HF in mice caused by CCl4 may be related to its ability to regulate liver hardness and inhibit the occurrence of epithelial mesenchymal transition in mice.
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Objective:To investigate the therapeutic effect and potential molecular mechanisms of cyclin-dependent kinase inhibitor-73 (CDKI-73), the Rab11 inhibitor, on liver fibrosis.Methods:Human LX2 cells were divided into four groups: negative control group, transforming growth factor-β (TGF-β) group, CDKI-73 group and TGF-β+ CDKI-73 group. Fifteen 5-week-old female C57 mice with body weight of (18.04±0.62) g were divided into 3 groups with 5 mice in each group: control group (intraperitoneal injection of olive oil + vehicle gavage), carbon tetrachloride (CCl 4) group (intraperitoneal injection of CCl 4 + vehicle gavage) and CCl 4+ CDKI-73 group (intraperitoneal injection of CCl 4+ CDKI-73 gavage). Another 15 5-week-old female C57 mice with body weight of (18.06±0.34) g were divided into 3 groups with 5 mice in each group: sham operation group (Sham), bile duct ligation (BDL) group + vehicle group (BDL+ vehicle gavage) and bile duct ligation+ CDKI-73 group (BDL+ CDKI-73 gavage). The expression of α-smooth muscle actin (α-SMA) and fibronectin(FN)in LX2 cells were analyzed by Western blot. Masson and Sirius red were used to examine the liver fibrosis after CDKI-73 treatment in vivo. Immunohistochemistry (IHC) was utilized to examine the expression of α-SMA in mice liver. Results:Collagen content assessed by Sirius red and Masson staining and α-SMA expression evaluated by IHC were all increased in CCl 4 group compared with control group ( q=38.47, 24.99, 36.79). Moreover, the collagen content and α-SMA expression in CCl 4 + CDKI-73 treatment group were obviously decreased compared with CCl 4 group ( q=24.72, 14.87, 27.50), and the differences were statistically significant (all P<0.001). Compared with Sham group, collagen content and α-SMA expression in bile duct ligation group were increased ( q=28.23, 41.01, 44.16). Furthermore, in BDL group, after treatment with CDKI-73, the collagen content and α-SMA expression were notably decreased ( q=22.88, 34.31 and 33.97, all P<0.001). Consistent with in vivo results, the relative expression levels of α-SMA and FN protein in TGF-β group were higher than those in TGF-β+ CDKI-73 group (α-SMA: 3.71±0.34 vs. 1.28±0.31; FN: 3.21±0.39 vs. 0.83±0.06, all P<0.001). The mRNA relative expression levels of α-SMA and FN in TGF-β group were higher than those in TGF-β+ CDKI-73 group, and the differences were statistically significant ( P<0.001). However, the relative expression of TGF-β receptor Ⅱ protein in CDKI-73 group was higher than those in negative control group (4.68±0.63 vs. 1.00±0.22, P=0.004). The relative expression level of phosphorylated SMAD2 in TGF-β+ CDKI-73 group was lower than those in TGF-β group (1.67±0.24 vs. 3.99±0.44, P<0.001). Transwell assay showed that 0.5 μmol/L CDKI-73 could effectively inhibit the migration of LX2 cells, and the inhibitory ability became stronger with the increase of CDKI-73 concentration. Conclusion:CDKI-73 can inhibit the activation of hepatic stellate cells and liver fibrosis by inhibiting Rab11-dependent TGF-β signaling pathway both in vivo and in vitro.
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Objective:To investigate the value of liver ultrasonic elasticity index combined with aspartate aminotransferase to platelet ratio index (APRI), fibrosis index based on the four factors (FIB-4) and globulin platelet model (GP) in the diagnosis of autoimmune hepatitis complicated with liver cirrhosis.Methods:From January 2020 to January 2022, 82 patients with autoimmune hepatitis and cirrhosis treated in West China Hospital of Sichuan University were selected as observation group, and 90 patients with autoimmune hepatitis were selected as controls (control group). All of them underwent liver ultrasound elastic examination, and the APRI, FIB-4, GP of patients were calculated. The differences of shear wave velocity (SWV), liver hardness value (LSM), strain rate ratio (SR), APRI, FIB-4, GP between the two groups were compared. At the same time, the differences of SWV, LSM, SR, APRI, FIB-4 and GP among patients with autoimmune hepatitis with different degrees of liver fibrosis and inflammation were analyzed. The value of liver ultrasound elasticity index, APRI, FIB-4 and GP in predicting autoimmune hepatitis complicated with cirrhosis was evaluated by the receiver operating characteristic (ROC) curve.Results:The SWV, LSM, FIB-4 and GP in the observation group were (1.60±0.21)m/s, (13.98±1.82)kPa, (8.10±1.43) and (4.15±1.05) respectively, which were significantly higher than those in the control group (all P<0.05), while SR and APRI were (5.04±0.98) and (2.41±0.92) respectively, which were significantly lower than those in the control group (all P<0.05). With the aggravation of liver fibrosis, the levels of SWV, LSM, FIB-4 and GP in patients with autoimmune hepatitis were higher (all P<0.05), while the SR and APRI were lower (all P<0.05). There was no statistically significant difference in SWV, LSM, SR, APRI, FIB-4 and GP between patients with G1-G2 and G3-G4 inflammatory degree of autoimmune hepatitis (all P>0.05). SWV, LSM, SR, APRI, FIB-4 and GP were included in the binary logistic regression analysis, and SWV, FIB-4 and GP were finally selected as independent predictors for diagnosis of autoimmune hepatitis with cirrhosis (all P<0.05). The area under the ROC curve of combined prediction of SWV, FIB-4 and GP for autoimmune hepatitis with cirrhosis was 0.931, which was significantly higher than other indicators (all P<0.05), and the sensitivity and specificity were 95.00% and 84.00% respectively. Conclusions:Liver ultrasonic elasticity index, APRI, FIB-4 and GP are related to the degree of liver fibrosis in patients with autoimmune hepatitis. SWV, FIB-4 combined with GP have high application value in predicting autoimmune hepatitis complicated with liver cirrhosis.
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Objective To investigate the role and mechanism of action of Scabiosa atropurea in inhibiting the proliferation of hepatic stellate cells using cell experiment. Methods A total of 20 Wistar rats were randomly divided into control group and administration group, with 10 rats in each group. The rats in the control group were given normal saline by gavage, and those in the administration group were given Scabiosa atropurea by gavage to prepare drug-containing serum. HSC-T6 cells were incubated with the serum from the control group (10%) or the low-, middle-, and high-dose serum containing Scabiosa atropurea (10%, 15%, and 20%, respectively). MTT assay was used to observe the effect of different drug concentrations on cells in different periods of time; flow cytometry was used to measure cell apoptosis; qRT-PCR and Western blot were used to measure the mRNA and protein expression levels of fibrosis markers (α-SMA, collagen Ⅰ) and PI3K/Akt signaling pathway-related factors in hepatic stellate cells (HSCs). A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t - test was used for further comparison between two groups. Results Compared with the control group, the low-, middle-, and high-dose serum containing Scabiosa atropurea groups had a significant reduction in the OD value of cells (all P < 0.05) and a significant increase in the overall apoptosis rate of cells (all P < 0.05). The results of qRT-PCR showed that compared with the control group, the low-, middle-, and high-dose serum containing Scabiosa atropurea groups had significant reductions in the mRNA expression levels of α-SMA, collagen Ⅰ, PI3K, and Akt and a significant increase in the mRNA expression level of PTEN (all P < 0.05); Western blot showed that compared with the control group, the low-, middle-, and high-dose serum containing Scabiosa atropurea groups had significant reductions in the protein expression levels of α-SMA, collagen Ⅰ, PI3K, Akt, and p-Akt and a significant increase in the protein expression level of PTEN (all P < 0.05). Conclusion The Mongolian medicine Scabiosa atropurea can inhibit the proliferation of HSC-T6 cells and promote their apoptosis, possibly by regulating fibrosis markers and the PI3K/Akt signaling pathway to exert an anti-liver fibrosis effect.
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Objective To investigate liver inflammation and fibrosis in patients with chronic HBV infection with a low viral load. Methods Among the HBsAg-positive patients who attended Ganzhou Fifth People's Hospital from April 2019 to June 2022, 41 patients with 20 IU/mL 200-2 000 IU/mL and 4 (26.67%) had an HBV DNA level of 20-200 IU/mL; the patients with G2 liver inflammation accounted for 73.33% (11/15), and those with S2 liver fibrosis accounted for 46.67% (7/15); the patients with ALT ≤30 U/L accounted for 46.67% (7/15); the patients aged ≥30 years accounted for 86.66% (13/15); the patients with LSM > 17 kPa accounted for 6.67% (1/15), those with an LSM value of 12.4-17 kPa accounted for 46.67% (7/15), and those with an LSM value of 9.4-12.4 kPa accounted for 33.33% (5/15). Conclusion Patients with chronic HBV infection and low-level viremia have a relatively high risk of progression of inflammation and fibrosis. ALT level cannot be used as a basis for antiviral therapy in HBV-infected people with a low viral load. LSM based on transient elastography can be used as a noninvasive test to screen for CHB patients with a low viral load, and antiviral therapy should be considered when HBV-infected patients with a low viral load have an age of > 30 years and an LSM value of > 9.4 kPa persistently.
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In recent years, significant breakthroughs have been made in the research on the role of traditional Chinese medicine (TCM) in the prevention and treatment of chronic hepatitis B liver fibrosis. In terms of clinical research, several multicenter randomized double-blind controlled studies have been conducted with liver pathological changes as the main efficacy evaluation indicator, which have improved the level of evidence in clinical research on the TCM prevention and treatment of liver fibrosis. As for basic research, in-depth studies have been conducted on the mechanism of action of TCM in intervening against liver fibrosis at various levels, including the effect on HSC activation, apoptosis and autophagy, inhibition of liver inflammatory injury, immunoregulation, and inhibition of hepatic sinusoidal capillarization, thereby highlighting the advantages and research difficulties of TCM through "multiple components, targets, and pathways". Under the guidance of Implementation Plan for the Major Projects of Revitalizing and Developing TCM, it is crucial to conduct high-quality clinical and basic research on TCM in the treatment of liver fibrosis, produce high-quality industry achievements, and thus help TCM to be recognized around the world.
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A new siderophore chelate (1) and 8 known compounds were identified from the liquid co-cultures of the marine-derived Streptomyces sp. IMB18-531 and Cladosporium sp. IMB19-099 by a combination of chromatography methods, including C18 reversed-phase medium pressure chromatography, gel column chromatography and HPLC. Their structures were determined by spectroscopic analysis and chemical methods as aluminioxamine E (1), desferrioxamine E (2), ferrioxamine E (3), terragine E (4), capsimicin (5), cyclo(L-prolinyl-L-tyrosine) (6), anthranilic acid (7), (Z)-14-methylpentadec-9-enoic acid (8), and (Z)-hexadec-8-enoic acid (9). Compound 2 showed inhibitory activities against the expression of liver fibrosis related genes COL1A1, MMP2, and TIMP2. Compounds 5, 8, and 9 displayed antibacterial activities against methicillin-resistant Staphylococcus aureus, S. epidermidis and Bacillus subtilis, with MICs of 16-64 μg·mL-1. Compound 5 showed cytotoxicities against human pancreatic cancer MIA Paca-2 and human colon cancer HT-29 cell lines with IC50 of 2.9 and 6.3 μmol·L-1, respectively.
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Objective To observe the effect of specific knockdown of hepatic stellate cells (HSC) ribosomal protein S5 (RPS5) on liver fibrosis in rats. Methods The glial fibrillary acidic protein (GFAP) promoter-driven RPS5 shRNA adenovirus was established, and AdGFa2-shRPS5 and its control AdGFa2 shNC were used to transfect primary rat HSCs and hepatocytes, respectively. RPS5 was determined by Western-blot and Real Time PCR, α-SMA and type I collagen expression; the rat liver fibrosis model was established by dimethyl nitrosamine (DMN) and bile duct ligation (BDL), and intrahepatic HSC was specifically knocked down by tail vein injection of adenovirus of RPS5 levels. The pathological changes of liver tissue sections were analyzed by HE staining; the content of hydroxyproline, sections of Sirius red and Masson staining were used to evaluate collagen deposition; immunohistochemical staining was used to detect the expression of α-SMA and RPS5. Results AdGFa2-shRPS5 specifically knocked down the expression level of RPS5 in HSC and increased the expression of α-SMA and type I collagen in vitro. The in vivo results showed that in two animal models of chronic liver injury, specific knockdown of RPS5 expression in HSCs promoted HSC activation, increased the deposition of extracellular matrix, and promoted liver fibrosis. Conclusion RPS5 is essential for HSC activation and liver fibrosis, which could be a potential target for the treatment of liver fibrosis.
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An effective therapeutic regimen for hepatic fibrosis requires a deep understanding of the pathogenesis mechanism. Hepatic fibrosis is characterized by activated hepatic stellate cells (aHSCs) with an excessive production of extracellular matrix. Although promoted activation of HSCs by M2 macrophages has been demonstrated, the molecular mechanism involved remains ambiguous. Herein, we propose that the vitamin D receptor (VDR) involved in macrophage polarization may regulate the communication between macrophages and HSCs by changing the functions of exosomes. We confirm that activating the VDR can inhibit the effect of M2 macrophages on HSC activation. The exosomes derived from M2 macrophages can promote HSC activation, while stimulating VDR alters the protein profiles and reverses their roles in M2 macrophage exosomes. Smooth muscle cell-associated protein 5 (SMAP-5) was found to be the key effector protein in promoting HSC activation by regulating autophagy flux. Building on these results, we show that a combined treatment of a VDR agonist and a macrophage-targeted exosomal secretion inhibitor achieves an excellent anti-hepatic fibrosis effect. In this study, we aim to elucidate the association between VDR and macrophages in HSC activation. The results contribute to our understanding of the pathogenesis mechanism of hepatic fibrosis, and provide potential therapeutic targets for its treatment.
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Humans , Hepatic Stellate Cells/pathology , Receptors, Calcitriol , Liver Cirrhosis/pathology , Macrophages/metabolismABSTRACT
Objective:To evaluate the efficacy of Qinggan Huashi Huoxue Decoction combined with conventional Western medicine in the treatment of alcoholic cirrhosis with spleen deficiency and phlegm stasis syndrome.Methods:Randomized controlled trial. A total of 110 patients from Tangshan Fengrun District Hospital of Traditional Chinese Medicine from March 2020 to March 2022 were selected as observation objects and divided into 2 groups with 55 patients in each group by computer random drawing method. The control group was treated with conventional Western medicine, while the observation group was treated with Qinggan Huashi Huoxue Decoction on the basis of the control group treatment. Both groups were treated for 3 months. The traditional Chinese medicine syndrome score was performed before and after treatment, and the levels of proline peptidase (PLD), type Ⅳ collagen (Ⅳ-C) and type Ⅰ procollagen aminopeptidase (PINP) were detected by phthalaldehyde contrast colorimetry, and the levels of pentamylin 3 (PTX3), protein kinase B (Akt) and B cell activating factor receptor (BAFF-R) were determined by ELISA. Adverse events were recorded and clinical efficacy was evaluated.Results:The total effective rate in the observation group was 92.73% (51/55), while that in the control group was 76.36% (42/55), the difference between the two groups was statistically significant ( χ2=5.64, P=0.018). After treatment, the score and total score of costal pain and fullness, swelling and firmness, anorexia, white and greasy tongue coating in the observation group were significantly lower than those in the control group ( t values were 11.02, 7.36, 7.47, 6.38, 9.37, respectively, P<0.01). After treatment, the levels of serum PLD[(143.28±16.38)U/L vs. (160.69±18.35)U/L, t=5.25], Ⅳ-C[(71.93±8.33)μg/L vs. (83.12±9.91)μg/L, t=6.41], and PINP[(32.36±5.32)ng/L vs. (39.02±5.61)ng/L, t=6.39] in the observation group were significantly lower than those in the control group ( P<0.01); The levels of PTX3[(36.82±4.96)ng/L vs. (42.14±5.83)ng/L, t=5.15], Akt[(69.22±7.94)ng/L vs. (77.24±8.63)ng/L, t=5.07], and BAFF-R[(15.29±3.64)ng/L vs. (19.92±4.15)ng/L, t=6.22] in the observation group were significantly lower than those in the control group ( P<0.01). During the treatment period, the incidence of adverse reactions was 12.73% (7/55) in the observation group and 9.09% (5/55) in the control group, with no statistically significant difference between the two groups ( χ2=0.37, P=0.541). Conclusion:Qinggan Huashi Huoxue Decoction combined with conventional Western medicine therapy can improve the Traditional Chinese Medicine syndrome and the degree of liver fibrosis damage in patients with alcoholic cirrhosis with spleen deficiency and phlegm stasis syndrome, inhibit the expression of serum inflammatory factors, and improve clinical efficacy.
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Objective:To study the mechanism of Herba Hedyotidis against liver fibrosis based on network pharmacology. Methods:Based on TCMSP database and Uniprot database, the effective components and target genes of Herba Hedyotidis were screened. Target genes of liver fibrosis were screened by GeneCards and OMIM database, and the "disease-component-target" network map was constructed by Cytoscape 3.8.2 software. Protein interaction network was constructed by STRING database, and the Cytoscape 3.8.2 software was used to screen the core target out. The core targets were analyzed by gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. Experimental verification was performed to the analysis results. A hepatic fibrosis model was established by intraperitioneal imjection of 40% carbon tetrachloride oil solution in rats that were then divided into the model control group and the Herba Hedyotidis group by randomized number table table, with 10 rats in each group. Ten normal rats were used as the normal control group. The Herba Hedyotidis group were injected 2.7 g/kg herb aqueous extract by intragastric administration, once a day, for 4 weeks; and the normal and model control group were given the same volume distilled water for gavage. The serum GPT, GOT, Alb and liver pathologic changes were observed. The serum expressions of IL-6, IL-1β and TGF-β1 were detected by ELISA. The expressions of PI3K, Akt, HIF-1α and VEGF were detected by Western blot. Results:5 effective components and 118 targets of Herba Hedyotidis in the treatment of hepatic fibrosis were obtained. Stigmasterol, β-sitosterol and quercetin were the most effective components with high moderate value. The moderate targets were VEGF, EGFR, HIF-1α and IL-6. The core genes of PPI network were HIF-1α, IL-6, etc. GO enrichment analysis showed that RNA transcription, protein binding and other processes may be affected. KEGG pathway enrichment analysis showed that significant enrichment pathways were cancer pathway, hepatitis B pathway, PI3K/Akt, HIF pathway and so on. Animal experimental results showed that compared with model group, liver histopathology was improved significantly, the content of GPT, GOT, IL-6, IL-1β and TGF-β1 decreased ( P<0.01), the content of Alb increased ( P<0.01), and the protein expressions of PI3K, Akt, HIF-1α and VEGF in liver tissue were down-regulated ( P<0.01). Conclusion:The Herba Hedyotidis exerts functions of anti-hepatic fibrosis through acting on the targets of VEGF, EGFR, HIF-1α and IL-6, regulating the PI3K/Akt, HIF-1 pathways, and has anti-inflammatory, anti-angiogenesis, anti-tumor and other biological functions.
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ObjectiveTo investigate the possible mechanism of Fugan Huaxian Decoction （扶肝化纤汤， FHD） against hepatic fibrosis （HF） from the perspective of immunity. MethodsForty-eight SD rats were randomly divided into blank group， model group， colchicine group， FHD high-， medium- and low-dose group， with eight rats in each group. Except for the blank group， the disease-syndrome combined model of HF with healthy qi deficiency and toxin accumulation pattern was established during six weeks in the other five groups. After successful modeling， the high-， medium- and low-dose FHD groups were respectively given 37.5， 18.75 and 9.38 g/（kg·d） of FHD granules by gavage， while the colchicine group received 2 mg/ （kg·d） of colchicine tablets by gavage， and the blank group and the model group were given 10 ml/（kg·d） of purified water， all for 3 weeks. The general condition of the rats was recorded. After the treatment， the histopathological morphology of the liver was observed by HE staining， and the levels of interleukin 10 （IL-10） and interleukin 17 （IL-17） in serum were determined by enzyme-linked immunosorbent assay （ELISA）. The expression of helper T cells 17 （Th17） and regulatory T cells （Treg） in peripheral blood were detected by flow cytometry， and the Th17/Treg value was calculated. The mRNA expression of retinoic acid-related nuclear orphan receptor γ （RORγt） and fork-head/wing-like helix transcription factor （FoxP3） in liver tissue were detected by qRT-PCR. ResultsCompared to the general condition of rats in the blank group， those in the model group were listless， less active， stretched and pushed， arched and prone， having no resistance to gavage， significantly reduced food intake， loose stools， dirty anus， slow weight gain， dry and dull hair， purple and darkening skin of the limbs with ecchymoses， purple and black spots with varying degrees of the skin of the tail； hepatic fibrosis and hyperplasia of rats in model group were more obvious； serum IL-17， peripheral blood Th17 expression and Th17/Treg value， RORγt mRNA expression in the liver tissue significantly increased in the model group， while expression of IL-10， Treg and FoxP3 mRNA significantly decreased （P＜0.05 or P＜0.01）. Compared to those in the model group， the general condition of the rats and the liver fibrosis of HE stained liver tissue were improved in all the medication groups； the expression of IL-17 and Th17， Th17/Treg， and RORγt mRNA expression significantly decreased， while expression of IL-10， Treg， and FoxP3 mRNA increased in the high- and medium-dose FHD groups and the colchicine group； the expression of IL-17， Th17， and RORγt mRNA decreased， while the expression of IL-10 and FoxP3 mRNA increased in the low-dose FHD group （P＜0.05 or P＜0.01）. And more improvements were found in the FHD high-dose group than FHD medium- and low-dose groups and colchicine group （P＜0.05 or P＜0.01）. ConclusionFHD can may regulate immune balance and act against fibrosis by regulating the expression of specific transcription factors FoxP3 and RORγt， affecting the differentiation of Th17 and Treg cells and Th17/Treg balance， and regulating the secretion of IL-10 and IL-17.
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The study aims to investigate the anti-hepatic fibrosis and anti-inflammatory activities of palbinone, and to explore the internal regulatory mechanism, so as to lay an active foundation for its development as an anti-non-alcoholic steatohepatitis (NASH) candidate. First, sulforhodamine B (SRB) method was used to detect the effect of palbinone on the proliferation of human hepatic stellate cells LX-2 and rat hepatic stellate cells HSC-T6. Following, in the in vitro hepatic fibrosis cell model that activated by transforming growth factor beta 1 (TGF-β1), quantitative real-time PCR (qRT-PCR) and Western blot were used to detect the inhibitory effect of different concentrations of palbinone on the transcription level and protein expression level of hepatic fibrosis markers. And the regulating mechanism of palbinone on fibrosis-related genes was analyzed at the same time. In addition, in the inflammatory cell model that induced by lipopolysaccharide (LPS) and nigericin, ELISA was used to detect the effect of palbinone on the released interleukin-1β (IL-1β) level. At the same time, Western blot was used to detect the effect of palbinone on the related proteins of inflammatory pathway. The results showed that palbinone could significantly inhibit the proliferation activity of LX-2 and HSC-T6, and their half maximal inhibitory concentration (IC50) values were (375.11 ± 55.45) and (260.27 ± 36.81) nmol·L-1, respectively. In addition, palbinone showed a dose-dependent inhibitory effect on the expression levels of TGF-β1-induced fibrosis-related genes, including collagen type Ⅰ α 1 (COL1A1), TGF-β1, α-smooth muscle actin (α-SMA) and tissue inhibitor of metalloproteinase 1 (TIMP1). Mechanism study showed that palbinone may decrease the expression level of Yes-associated protein (YAP), thereby weakening its activation effect on the downstream fibrosis pathway. In addition, palbinone also exerted an anti-inflammatory effect by inhibiting the activity of nuclear factor kappa-B (NF-κB) signaling pathway and reducing inflammatory factors cysteinyl aspartate specific proteinase-1 (caspase-1) and IL-1β release. In conclusion, palbinone can not only inhibit the proliferation and activation of hepatic stellate cells by inhibiting the expression of YAP, but also inhibit the expression and release of inflammatory factors at the same time. All these studies provide theoretical support for the development of palbinone as an anti-nonalcoholic steatohepatitis drug.
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OBJECTIVES@#Hepatic fibrosis is a serious pathological consequence of chronic liver disease. Mycophenolate mofetil (MMF) is a commonly used immunosuppressant after organ transplant. However, the relationship between MMF and hepatic fibrosis remains unclear. This study aims to explore the effect of MMF on hepatic fibrosis in mice and the potential mechanism.@*METHODS@#A total of 24 mice (male, 8-week old, C57BL/6) were randomly divided into a control group, a MMF group, a carbon tetrachloride (CCl4) group and a CCl4+MMF group (n=6 in each group). After the mice were sacrificed, the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were detected. The liver tissues were taken up for Masson staining and collagen I (COL1) immunohistochemistry. The levels of transforming growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) were detected by Western blotting. Finally, the levels of mRNA for TGF-β1, α-SMA, and COL1 were detected using real-time PCR.@*RESULTS@#Compared with the CCl4 group, the ALT and AST levels were lower (both P<0.05), the degree of liver fibrosis was alleviated, and the deposition of COL1 in the liver was significantly decreased (P<0.01) in the CCl4+MMF group. Compared with the CCl4 group, the protein expression levels of TGF-β1 and α-SMA were significantly decreased (both P<0.05) and the relative expression levels of TGF-β1, α-SMA and COL1 mRNA in the liver were significantly decreased (all P<0.05) in the CCl4+MMF.@*CONCLUSIONS@#MMF could reduce CCl4-induced hepatic fibrosis, which might be related to the inhibition of TGF-β1. This study is expected to provide a target for the treatment of hepatic fibrosis.