基于JAK2/STAT3信号通路探讨舒肝化癥方对大鼠肝纤维化的作用机制
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1.甘肃中医药大学;2.甘肃中医大学;3.甘肃中医药大

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Effect of Shugan Huazheng-Fang powder on liver fibrosis in rats based on JAK2/STAT3 signaling pathway
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Gansu University of Chinese Medicine

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    摘要:

    目的:探讨舒肝化癥方在大鼠肝纤维化模型及肝星状细胞(HSC-T6)中的作用及机制。方法:54只雄性SD大鼠随机分为空白组、模型组及低剂量(7.38g/kg)、中计量(14.76g/kg)和高剂量(29.52g/kg)舒肝化癥方组及秋水仙碱组(0.2mg/kg),每组8只。模型组及各给药组大鼠腹腔注射40%四氯化碳(CCl4)溶液,同时各给药组每天予以相应剂量的药物灌胃8周。采用全自动生化分析仪检测肝血清生化指标ALT、AST;HE和天狼星染色观察肝组织损伤情况;ELISA检测肝组织中α-SMA、Col Ⅰ表达水平;Western blot检测肝组织中JAK2、P-JAK2、STAT3、P-STAT3、IL-6、IL-1β、TNF-α蛋白表达水平;RT-qPCR检测肝组织中JAK2、STAT3、IL-6、IL-1β、TNF-α mRNA表达水平。将HSC-T6细胞分为空白组、模型组、含药血清组、抑制剂组及联合组,除空白组外均用LPS诱导,CCK-8法检测细胞活力;ELISA法检测α-SMA、Col Ⅰ表达水平;Western blot检测细胞中JAK2、P-JAK2、STAT3、P-STAT3、IL-6、IL-1β、TNF-α蛋白表达水平;RT-qPCR检测细胞中JAK2、STAT3、IL-6、IL-1β、TNF-α mRNA表达水平。结果:舒肝化癥方剂量依赖性改善肝功能,并能减轻肝纤维化程度;与模型组比较,阳性对照组及舒肝化癥方各剂量组JAK2/STAT3信号通路及炎症相关因子表达均显著降低(P<0.05);舒肝化癥方含药血清显著下调LPS诱导的HSC-T6细胞中α-SMA、Col Ⅰ、JAK2/STAT3信号通路及炎症相关因子表达水平(P<0.05)。结论:舒肝化癥方在大鼠体内、体外均有抗肝纤维化的作用,该作用机制与JAK2/STAT3信号通路及炎症的表达有关。

    Abstract:

    Objective:To investigate the effects and mechanisms of Shugan Huazheng Decoction in rat liver fibrosis model and hepatic stellate cells (HSC-T6). Method:Fifty-four male SD rats were randomly divided into blank group, model group and low-dose (7.38g/kg), medium-dose (14.76g/kg) and high-dose (29.52g/kg) Shugan Huazheng Decoction group and colchicine group (0.2mg/kg), with 8 rats in each group. Rats in the model group and each dosing group were injected intraperitoneally with 40% carbon tetrachloride (CCl4) solution, while each dosing group was given the corresponding dose by gavage daily for 8 weeks. The liver serum biochemical indexes ALT and AST were detected by fully automatic biochemical analyzer; HE and Sirius staining was used to observe the liver tissue injury; ELISA detected the expression levels of α-SMA and Col Ⅰ in liver tissue; Western blot detected the protein expression levels of JAK2, P-JAK2, STAT3, P-STAT3, IL-6, IL-1β, and TNF-α; and RT-qPCR detected the mRNA expression levels of JAK2, STAT3, IL-6, IL-1β, and TNF-α in liver tissue. HSC-T6 cells were divided into blank group, model group, drug-containing serum group, inhibitor group and combined group, except blank group, all were induced by LPS, cell viability was detected by CCK-8; α-SMA, Col Ⅰ expression level was detected by ELISA; JAK2, P-JAK2, STAT3, P-STAT3, IL-6, IL-1β, TNF-α protein expression level in cells was detected by western blot JAK2, P-JAK2, STAT3, P-STAT3, IL-6, IL-1β, and TNF-α protein expression levels in cells; RT-qPCR detection of JAK2, STAT3, IL-6, IL-1β, and TNF-α mRNA expression levels in cells. Result:Shugan Huazheng Decoction dose-dependently improved liver function and attenuated hepatic fibrosis; the expression of JAK2/STAT3 signaling pathway and inflammation-related factors were significantly reduced in the positive control group and all Shugan Huazheng Decoction dose groups compared with the model group(P<0.05);Shugan Huazheng Decoction -containing serum significantly down-regulated the expression levels of α-SMA, Col Ⅰ, JAK2/STAT3 signaling pathway and inflammation-related factors in LPS-induced HSC-T6 cells(P<0.05).Conclusion:Shugan Huazheng Decoction showed anti-hepatic fibrosis effects in rats in vivo and in vitro, and the mechanism of action was related to the expression of JAK2/STAT3 signaling pathway and inflammation.

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  • 收稿日期:2024-03-22
  • 最后修改日期:2024-09-26
  • 录用日期:2024-12-02
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