AIM:To study the differential expression of proteins between natural taurine treated hepatic stellate cells and controls, and investigate the underlying regulatory mechanism of natural taurine in inhibiting hepatic fi...AIM:To study the differential expression of proteins between natural taurine treated hepatic stellate cells and controls, and investigate the underlying regulatory mechanism of natural taurine in inhibiting hepatic fibrosis.METHODS: A proteomic strategy combining two-dimensional gel electrophoresis and ultraperform ance liquid chromatographyelectrospray ionizationtandem mass spectrometry (UPLCESIMS/MS) was used to study the differential expression of proteins and Western blotting was used to validate the results. Gene ontology (GO) method was utilized to analyze the functional enrichment of differentially expressed proteins. Flow cytometry was performed to compare the apoptosis rate between taurinetreated and untreated hepatic stellate cells (HSCs).RESULTS: Nineteen differentially expressed proteins (11 upregulated and 8 downregulated) were identifiedby 2D/MS, and the expression profiles of GLO1 and ANXA1 were validated by Western blotting. GO analysis found that these differentially expressed proteins were enriched within biological processes such as "cellular apoptosis", "oxidation reaction" and "metabolic process" in clusters. Flow cytometric analysis showed that taurinetreated HSCs had a significantly increased apoptosis rate when compared with the control group.CONCLUSION: Natural taurine can promote HSC apoptosis so as to inhibit hepatic fibrosis.展开更多
AIM To develop a reliable and simple method to identify important biological metabolites and relevant pathways for taurine in hepatic stellate cells(HSCs), in order to provide more data for taurine therapy.METHODS All...AIM To develop a reliable and simple method to identify important biological metabolites and relevant pathways for taurine in hepatic stellate cells(HSCs), in order to provide more data for taurine therapy.METHODS All the biological samples were analyzed by using highperformance liquid chromatography-time electrospray ionization/quadrupole-time of flight mass spectrometry. Principal component analysis and partial least squares discriminant analysis were used to identify statistically different metabolites for taurine in HSCs, and metabolomic pathway analysis was used to do pathway analysis for taurine in HSCs. The chemical structure of the related metabolites and pathways was identified by comparing the m/z ratio and ion mode with the data obtained from free online databases.RESULTS A total of 32 significant differential endogenous metabolites were identified, which may be related to the mechanism of action of taurine in HSCs. Among the seven relevant pathways identified, sphingolipid metabolism pathway, glutathione metabolism pathway and thiamine metabolism pathway were found to be the most important metabolic pathways for taurine in HSCs.CONCLUSION This study showed that there were distinct changes in biological metabolites of taurine in HSCs and three differential metabolic pathways including sphingolipid pathway, glutathione pathway and thiamine metabolism pathway might be of key importance in mediating the mechanism of action of taurine in HSCs.展开更多
基金Supported by The National Natural Science Foundation of China, No. 30660235Guangxi Natural Science Foundation, No. 0728080
文摘AIM:To study the differential expression of proteins between natural taurine treated hepatic stellate cells and controls, and investigate the underlying regulatory mechanism of natural taurine in inhibiting hepatic fibrosis.METHODS: A proteomic strategy combining two-dimensional gel electrophoresis and ultraperform ance liquid chromatographyelectrospray ionizationtandem mass spectrometry (UPLCESIMS/MS) was used to study the differential expression of proteins and Western blotting was used to validate the results. Gene ontology (GO) method was utilized to analyze the functional enrichment of differentially expressed proteins. Flow cytometry was performed to compare the apoptosis rate between taurinetreated and untreated hepatic stellate cells (HSCs).RESULTS: Nineteen differentially expressed proteins (11 upregulated and 8 downregulated) were identifiedby 2D/MS, and the expression profiles of GLO1 and ANXA1 were validated by Western blotting. GO analysis found that these differentially expressed proteins were enriched within biological processes such as "cellular apoptosis", "oxidation reaction" and "metabolic process" in clusters. Flow cytometric analysis showed that taurinetreated HSCs had a significantly increased apoptosis rate when compared with the control group.CONCLUSION: Natural taurine can promote HSC apoptosis so as to inhibit hepatic fibrosis.
基金Supported by National Natural Science Foundation of China,No.81360595 and No.81360532Guangxi Natural Science Foundation Program,No.2014GXNSFDA118027+1 种基金Bagui Scholars Foundation Program of GuangxiSpecial-term Experts Foundation Program of Guangxi
文摘AIM To develop a reliable and simple method to identify important biological metabolites and relevant pathways for taurine in hepatic stellate cells(HSCs), in order to provide more data for taurine therapy.METHODS All the biological samples were analyzed by using highperformance liquid chromatography-time electrospray ionization/quadrupole-time of flight mass spectrometry. Principal component analysis and partial least squares discriminant analysis were used to identify statistically different metabolites for taurine in HSCs, and metabolomic pathway analysis was used to do pathway analysis for taurine in HSCs. The chemical structure of the related metabolites and pathways was identified by comparing the m/z ratio and ion mode with the data obtained from free online databases.RESULTS A total of 32 significant differential endogenous metabolites were identified, which may be related to the mechanism of action of taurine in HSCs. Among the seven relevant pathways identified, sphingolipid metabolism pathway, glutathione metabolism pathway and thiamine metabolism pathway were found to be the most important metabolic pathways for taurine in HSCs.CONCLUSION This study showed that there were distinct changes in biological metabolites of taurine in HSCs and three differential metabolic pathways including sphingolipid pathway, glutathione pathway and thiamine metabolism pathway might be of key importance in mediating the mechanism of action of taurine in HSCs.
文摘目的通过观察天然牛磺酸(NT)对肝纤维化大鼠肿瘤坏死因子(TNF)-α表达的影响,研究NT抑制肝细胞炎性状态、延缓肝纤维化进展的相关机制。方法将实验大鼠随机分为对照组、模型组、NT低剂量组(0.3 g/kg)、NT中剂量组(0.6 g/kg)、NT高剂量组(0.9 g/kg),每组15只。对照组给予正常喂养;模型组与NT各组大鼠均给予皮下注射40%CCL4橄榄油溶液3 m L/kg造模,首次剂量5 m L/kg,2次/w,共注射6 w,并饲以高脂饲料;NT各组在造模的同时予天然牛磺酸灌胃,连续给药6 w;实验结束时,称量大鼠体质量和肝脏质量,计算肝体系数,检测大鼠肝功能(ALT、AST、TBIL及ALB)、肝纤维化指标(HA、PCIII、IV-C及LN)、血清TNF-α水平及肝脏组织TNF-αmRNA表达量。结果与模型组相比,NT低剂量组肝体系数差异无统计学意义(P>0.05),NT中剂量组、高剂量组肝体系数明显降低;NT各组肝功能、肝纤维化显著改善,血清TNF-α水平及肝脏组织中TNF-αmRNA表达显著降低。结论NT能维持肝功能、延缓肝纤维化进展,其机制可能与抑制TNF-α相关因子表达,降低炎性介质分泌,减轻炎症反应有关。
