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目的观察铁包金对慢加急性肝衰竭的作用及其对无嘌呤/无嘧啶核酸内切酶1(APE1)、caspase3、bax、bcl-2的影响。方法取SD大鼠40只,随机分为4组,每组10只,分别为正常对照组、模型组、铁包金组(8 g·kg~(-1)·d~(-1))和安宫牛黄丸组(0.27 g·kg~(-1)·d~(-1))。采用四氯化碳(CCL4)联合D-氨基半乳糖(D-Gal N)和脂多糖(LPS)复制慢加急性肝衰竭模型。观察大鼠48 h内的病死率。全自动生化仪检测丙氨酸氨基转移酶(ALT)、天冬氨酸转移酶(AST)、血清总胆红素(TBIL)的水平。HE染色观察肝脏病理变化。Tunel染色观察肝细胞凋亡指数(AI)。免疫印迹检测APE1、caspase3、bax和bcl-2蛋白的表达。共聚焦显微镜观察APE1的原位表达。结果正常对照组、模型组、铁包金组和安宫牛黄丸组48 h病死率分别为0%、50%、20%和20%。与模型组比较,铁包金组显著降低ALT、AST、TBIL、AI、caspase3和bax的水平,差异有统计学意义(P<0.05);铁包金组可显著提高APE1、bcl-2的水平,差异有统计学意义(P<0.05);铁包金组作用效果与安宫牛黄丸组相当,差异无统计学意义(P>0.05)。结论铁包金对慢加急性肝衰竭具有一定的防治作用,可能是铁包金通过APE1/Ref~(-1)调控凋亡相关蛋白防治慢加急性肝衰竭。
Objective To observe the effects of ferrules on chronic and acute liver failure and its effects on apoptin, pyrimidine endonuclease 1 (APE1), caspase3, bax and bcl-2. Methods Forty Sprague-Dawley rats were randomly divided into 4 groups (n = 10): normal control group, model group, Fe-Au group (8 g · kg -1 · d -1) And Angong Niuhuang Wan group (0.27 g · kg -1 · d -1). Acute hepatic failure model was replicated with CCL4 combined with D-Gal N and LPS. The mortality of rats within 48 h was observed. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total serum bilirubin (TBIL) were detected by automatic biochemical analyzer. HE staining was used to observe the pathological changes of liver. Tunel staining observed hepatocyte apoptosis index (AI). Western blot was used to detect the expression of APE1, caspase3, bax and bcl-2 protein. Confocal microscopy was used to observe the expression of APE1 in situ. Results The 48 h mortality rates of normal control group, model group, iron-coated group and Angong Niuhuang Wan group were 0%, 50%, 20% and 20% respectively. Compared with the model group, the Fe-Au group significantly reduced the levels of ALT, AST, TBIL, AI, caspase3 and bax, the difference was statistically significant (P <0.05) , The difference was statistically significant (P <0.05). The effect of iron-coated group was similar to that of Angong Niuhuang Wan group, with no significant difference (P> 0.05). CONCLUSION: Iron-coated gold may have some preventive and therapeutic effects on chronic and acute liver failure. It may be that iron-coated gold protects against acute liver failure by APE1 / Ref ~ (-1) regulation of apoptosis-related proteins.