截断逆挽方对慢加急性肝衰竭大鼠死亡受体途径的影响

Effect of JieDuan NiWan prescription on death receptor pathway in acute-on-chronic liver failure rat model

目的观察"截断逆挽方"对慢加急性肝衰竭大鼠JNK信号通路死亡受体途径的影响。方法 SPF级Wistar雄性大鼠70只,随机分为正常组、模型组、截断逆挽方组和SP600125组,各组大鼠给予猪血清腹腔注射13周,联合D氨基半乳糖/脂多糖(D-galactosamine/lipopolysaccharide,D-Gal N/LPS)急性攻击,构建慢加急性肝衰竭大鼠模型,截断逆挽方组在急性攻击前给予截断逆挽方连续灌胃3天,SP600125组于急性攻击前半小时腹腔注射。各组大鼠均在急性攻击后4、8、12小时平行取材。用免疫组化法(Immunohistochemical detection,IHC)检测肝组织中半胱氨酸蛋白酶-8(Cysteine Proteinase-8,Caspase-8)、Caspase-3、脂肪酸合成酶(Fatty acid synthase,Fas)的表达量,用蛋白印迹法(Western Blot,WB)检测肝组织中Fas配体(Fas ligand,FasL)的表达量。结果与正常组相比,模型组各时间点Caspase-3、Caspase-8、Fas、Fas L表达增多,差异有统计学意义(P<0.05);与模型组相比,截断逆挽方组及SP600125组在4小时和8小时表达减少,差异有统计学意义(P<0.01或P<0.05),12小时较模型组升高,差异无统计学意义(P>0.05)。结论截断逆挽方可以有效降低Caspase-3、Caspase-8、Fas、FasL的表达量,阻断死亡受体途径,减少模型大鼠肝细胞凋亡。

Objective To observe the effect of Jieduan Niwan （JDNW） on the death receptor pathway of JNK signaling pathways in acute-on-chronic liver failure （ACLF） rats, and study its mechanism. Methods 70 SPF male Wistar rats were randomly divided into normal group, model group, Jieduan-Niwan prescription group and JNK inhibitor SP600125 group. Except normal group, other groups were given intraperitoneal injection with porcine serum for 13 weeks combined with D-galactosamine / lipopolysaccharide （D-GalN/LPS） acute attack to induce the model of ACLF, JDNW group intragastric ad- ministration for 3 days before acute attack, SP600125 group was intraperitoneal injection half an hour before acute attack. The rats were sacrificed in 4,8,12h after acute attack. Caspase-3, Caspase-8, Fas in liver tissue was detected by immunohistochemical method, the expression of FasL in liver tissue was detected by Western Blot. Results Compared with normal group, the expression of Caspase-3, Caspase-8, Fas, FasL were increased （P〈 0.05 ） in model group at different time points; compared with the model group, JDNW group and SP600125 group was reduced at 4h and 8h（ P〈0. 01 or P〈0.05 ）, while that was higher than model group at 12h, result had not statistically significant （P〉0. 05）. Both JDNW group and SP600125 group can reduce the expression of Caspase-3, Caspase-8, Fas, FasL, and the mechanism was similar. Conclusion The prescription of Jieduan-Niwan can effectively reduce the expression of Caspase-3, caspase-8, Fas, FasL, the mechanism may be related to blocking the death receptor pathway of JNK signaling pathway.