大承气汤通过调节TGF-β1/Smad3信号通路减轻大鼠急性肝损伤及全身炎症反应

Dachengqi Decoction Protects Acute Liver Injury and Systemic Inflammatory Response by Regulating TGF-β1/Smad3 signaling pathway

目的观察大承气汤对盲肠结扎穿刺(cecum ligation and puncture,CLP)诱导的脓毒症大鼠急性肝损伤的保护作用及对全身炎症的影响,并探讨其可能的作用机制。方法30只雄性SPF级SD大鼠(Sprague-Dawley rats)随机分为假手术对照组、模型对照组、大承气汤2.5 g/kg组、大承气汤5 g/kg组、大承气汤10 g/kg组5个小组,每组6只(n=6)。运用苏木精-伊红染色(hematoxylin-eosin staining,HE)观察大鼠肝组织病理学改变,检测大鼠肝脏组织中天冬氨酸转氨酶(aspartate transaminase,AST)和丙氨酸转氨酶(alanine transaminase,ALT)的变化;采用Elisa检测大鼠血清中促炎细胞因子IL-1β、IL-6、TNF-α的水平;通过免疫组织化学(IHC)观察大鼠肝脏中转化生长因子-β1(TGF-β1)和Smad3的蛋白表达差异;最后运用免疫印迹试验(Western Blot)检测肝脏组织中TGF-β1和Smad3的蛋白表达情况。结果与假手术对照组相比,模型对照组大鼠肝脏组织病理损伤明显,肝功能显著上升(P<0.01),大鼠全身促炎细胞因子分泌显著增加(P<0.01),转化生长因子-β1(TGF-β1)和Smad3的蛋白表达显著上升(P<0.01);与模型对照组相比,大承气汤给药后,肝脏组织病理损伤逐渐减轻,肝功能显著下降(P<0.01),大鼠全身促炎细胞因子分泌明显降低(P<0.05),转化生长因子-β1(TGF-β1)和Smad3的蛋白表达显著下降(P<0.01),且呈浓度梯度。结论大承气汤可以对盲肠结扎穿刺诱导的脓毒症相关的急性肝损伤发挥保护作用,改善其肝功能,抑制大鼠全身炎症反应,其作用机制可能通过调节TGF-β1/Smad3信号通路实现。

Objective Effects of Dachengqi decoction on acute liver injury induced by cecal puncture and ligation(CLP)and systemic inflammation were observed. Methods 30 male SPF grade SD rats were randomly divided into control group(Control),model group(Model), low concentration group of Dachengqi decoction(DCQD L), medium concentration group of Dachengqi decoction(DCQD M), high concentration group of Dachengqi decoction(DCQD H), DCQD L group was given 2.5 g/kg Dachengqi decoction(i.g,bid), DCQD M group was given 5 g/kg Dachengqi decoction(i.g,bid) and DCQD H group was given 10 g/kg Dachengqi decoction, 6 in each group. Histopathological changes in liver of rats were observed by hematoxylin-eosin staining, The difference of TGF-β1 and Smad3 expression in liver of rats were observed by immunohistochemistry;The changes of aspartate aminotransferase and alanine aminotransferase in rat liver were detected;The levels of pro-inflammatory cytokines interleukin 1β, interleukin 6 and tumor necrosis factor α in rats serum were detected by enzyme-linked immunosorbent assay;finally, Westen blot was used to detect the protein expression of TGF-β1 and Smad3 in liver. Results CLP could induce acute liver injury associated with sepsis compared with Control group, resulting in obvious liver pathological injury and abnormal liver function and increase the secretion of proinflammatory cytokines in rats. Dachengqi decoction could improve liver function and liver pathological injury, and it could reduce the systemic inflammatory reaction, reduce the protein expression of TGF-β1 and Smad3,and showed a concentration gradient.Conclusion Dachengqi decoction plays a protective role by CLP induced sepsisrelated acute liver injury, improves its liver function and inhibits the systemic inflammatory response in rats. Its mechanism may be realized by regulating TGF-β1/Smad3 signaling pathway.