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目的本实验以细胞模型为基础,通过western blot方法检测水通道蛋白AQP-4及occludin蛋白在肝硬化肝性脑病脑水肿脑组织中的表达,同时结合脑组织病理形态学的变化,来探讨毒消肝清丸对实验大鼠肝硬化肝性脑病脑水肿的影响,从而进一步证实毒消肝清丸对肝性脑病的治疗作用及机理。方法用四氯化碳、注射硫代乙酰胺联合造模制备肝硬化肝性脑病模型,模型组给予蒸馏水灌胃;高剂量组、中剂量组、低剂量组分别给予毒消肝清丸(223.4mg/kg)、(111.7mg/kg)、(58.9mg/kg)日一次灌胃给药,连续给药7天;对照组给予乳果糖(配置浓度100g/L,以1ml/100g灌胃)日一次灌胃给药,连续给药7天。用HE染色观察脑组织病理形态学的改变;用Western-Blot方法检测大鼠脑组织中occludin与AQP-4蛋白的表达情况。结果乳果糖组、高剂量、中剂量和低剂量试验药组肝性脑病脑水肿程度明显减轻,但以中、高剂量更为明显;AQP-4及occludin蛋白的western blot方法检测分析显示:乳果糖组、毒消肝清丸低、中、高剂量组较模型组相比,有显著性差异(P<0.05)。结论毒消肝清丸从补气健脾、滋阴清热、泻毒导滞入手,具有清肠毒、通宿便、减少肠道内毒素吸收、降低肝损害、改善血脑屏障和肠道屏障的功能,可达到防治肝性脑病的目的。
ObjectiveTo investigate the expression of aquaporin AQP-4 and occludin protein in cerebral edema induced by hepatic cirrhosis and liver cirrhosis by Western blot, Heganqing pill on liver cirrhosis in rats with cirrhosis of brain edema, to further confirm the toxic Xiaoheqing pill on hepatic encephalopathy and the mechanism of action. Methods Hepatic cirrhosis model was induced by carbon tetrachloride and thioacetamide injection. The model group was administered with distilled water. The high dose group, the medium dose group and the low dose group were administered with Tangxueganqing pill (223.4mg (kg / kg), (111.7mg / kg), (58.9mg / kg) once daily intragastric administration, continuous administration of 7 days; control group was given lactulose (configuration concentration 100g / L, Once intragastric administration, continuous administration for 7 days. The histopathological changes of brain tissue were observed by HE staining. The expression of occludin and AQP-4 protein in brain tissue were detected by Western-Blot. Results The levels of cerebral edema in hepatic encephalopathy of lactulose group, high dose, middle dose and low dose test group were significantly reduced, but were more obvious in middle and high dose group. Western blot analysis of AQP-4 and occludin protein showed that milk Compared with the model group, there were significant differences in the fructose group and the drug-treated Ganganqing pill in the low, medium and high dose groups (P <0.05). CONCLUSION DDP has the functions of invigorating qi and spleen, nourishing yin and clearing heat and purging toxins. It has the functions of clearing body intestine, reducing the absorption of intestinal endotoxin, reducing liver damage, improving blood-brain barrier and intestinal barrier, To achieve the purpose of prevention and treatment of hepatic encephalopathy.