双参饮对肝纤维化模型大鼠的改善作用及机制研究
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篇名: 双参饮对肝纤维化模型大鼠的改善作用及机制研究
TITLE:
摘要: 目的:研究双参饮对肝纤维化模型大鼠的肝功能和肝纤维化程度的改善作用及其机制。方法:取健康SD大鼠,以含40%四氯化碳的橄榄油溶液连续7周腹腔注射,建立肝纤维化模型。将造模成功的47只大鼠随机分为模型组、阳性对照组和双参饮低、中、高剂量组,各组分别为9、9、9、10、10只;另取10只健康SD大鼠作为正常组。从造模结束后次周开始,模型组与正常组大鼠灌胃水,阳性对照组[秋水仙碱0.2 mg/(kg·d)]和双参饮各剂量组[双参饮水煎液3、6、12 g/(kg·d),以总生药量计算]大鼠分别灌胃相应药液,连续给药28 d。采用赖氏分析法检测血清中肝功能指标[丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆红素(TBIL)、白蛋白(ALB)、球蛋白(GLB)]水平,采用酶联免疫吸附法检测血清中肝纤维化指标[透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型胶原(Ⅲ-PC)、Ⅳ型胶原前体(Ⅳ-C)]水平;采用苏木精-伊红染色法和苦味酸酸性复红染色法观察大鼠肝组织病理学变化;采用分光光度法检测大鼠肝组织中超氧化物歧化酶(SOD)、丙二醛(MDA)、羟脯氨酸(HYP)水平;采用免疫组化法检测大鼠肝组织中α-平滑肌肌动蛋白(α-SMA)表达水平。结果:与正常组比较,模型组大鼠血清中ALT、AST、TBIL、HA、LN、PC-Ⅲ、Ⅳ-C水平均显著升高,ALB水平显著降低(P<0.05);大鼠肝组织出现空泡变性、纤维组织增生等病理现象;肝组织中SOD水平显著降低,MDA、HYP水平均显著升高(P<0.05);肝组织中可见大量α-SMA蛋白阳性染色颗粒,其蛋白表达水平显著升高(P<0.05)。与模型组比较,双参饮各剂量组大鼠血清中ALT、AST和肝纤维化指标水平均显著降低,ALB水平显著升高,双参饮中、高剂量组TBIL水平均显著降低(P<0.05);肝细胞变性、纤维化组织增生现象均有较大改善;肝组织中SOD水平显著升高,MDA、HYP水平均显著降低(P<0.05);α-SMA蛋白阳性染色明显减少,其蛋白表达水平显著降低(P<0.05)。结论:双参饮对肝纤维化模型大鼠有保肝、抗肝纤维化的作用,其机制可能主要是通过增强机体抗氧化能力、下调α-SMA蛋白表达、抑制肝星状细胞的活化、减少细胞外间质的合成,从而逆转肝纤维化进程。
ABSTRACT: OBJECTIVE: To study improvement effect and mechanism of Shuangshen yin on liver function and the degree of liver fibrosis in liver fibrosis model rats. METHODS: Healthy SD rats were collected and given 40% CCl4 olive oil solution intraperitoneally for consecutive 7 weeks to induce liver fibrosis model. 47 model rats were randomly divided into model group, positive control group and Shuangshen yin low-dose, medium-dose and high-dose groups, with 9, 9, 9, 10, 10 rats respectively; another 10 healthy SD rats were included in normal group. From the next week after model induction, model group and normal group were given water intragastrically; positive control group [colchicines 0.2 mg/(kg·d)] and Shuangshenyin groups [Shuangshen yin water decoction 3, 6, 12 g/(kg·d), by crude drug] were given corresponding medicines respectively for consecutive 28 d. The serum levels of liver function indicators (ALT, AST, TBIL, ALB, GLB) were determined by Rye’s analysis method. The serum levels of liver fibrosis indexes (HA, LN, Ⅲ-PC, Ⅳ-C) were determined by ELISA. The pathological changes of liver tissues were observed by HE staining and VG staining. The levels of SOD, MDA and HYP in liver tissues were determined by spectrophotometry. The expression of α-SMA was detected by immunohistochemistry. RESULTS: Compared with normal group, the serum levels of ALT, AST, TBIL, HA, LN, PC-Ⅲ and Ⅳ-C in model group were increased significantly, and the level of ALB was decreased significantly (P<0.05). There were some pathological phenomena such as vacuolar degeneration and fibrous tissue proliferation in rat liver. The levels of SOD in liver tissue were decreased significantly, while the levels of MDA and HYP were increased significantly (P<0.05). A large number of positive staining granules of α-SMA protein were observed in liver tissue, and the expression level of α-SMA protein was significantly increased (P<0.05). Comparison of model group, the serum levels of ALT, AST and liver fibrosis indexes were significantly decreased and ALB level was significantly increased in Shuangshen yin groups, while the TBIL levels in Shuangshen yin medium-dose, high-dose groups were significantly decreased (P<0.05); the phenomena of hepatocyte degeneration and fibrosis tissue proliferation were significantly improved; the levels of SOD in liver tissue were significantly increased, and the levels of MDA and HYP were significantly decreased (P<0.05); the positive staining of α-SMA protein decreased significantly, and the expression level of α-SMA protein was decreased significantly (P<0.05). CONCLUSIONS: Shuangshen yin has protective and anti-fibrotic effects on liver in rats with hepatic fibrosis, the mechanism of which may be associated with enhancing antioxidant capacity, down-regulating the expression of α-SMA, inhibiting the activation of hepatic stellate cells, reducing extracellular matrix synthesis so as to reverse liver fibrosis.
期刊: 2019年第30卷第12期
作者: 黄祎,周刚,钟珊,周智,扈晓宇
AUTHORS: HUANG Yi,ZHOU Gang,ZHONG Shan,ZHOU Zhi,HU Xiaoyu
关键字: 双参饮;丹参;人参;肝纤维化;机制;大鼠
KEYWORDS: Shuangshen yin; Salvia miltiorrhiza; Panax schinseng; Liver fibrosis; Mechanism; Rats
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