左归饮对D-半乳糖致衰大鼠肝肾组织形态及其细胞凋亡相关蛋白表达的影响

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目的研究左归饮对D-半乳糖致亚急性衰老模型大鼠肝、肾组织形态及其细胞凋亡相关蛋白表达的影响,初步探讨其延缓衰老作用机制。方法取健康SD大鼠72只,雌雄各半,随机分为6组,即对照组、模型组、给药组(维生素E组,左归饮高、低剂量组和六味地黄丸组)。除对照组颈背部皮下注射0.9%Na Cl溶液外,其余各组颈背部皮下分别连续注射D-半乳糖建立亚急性衰老模型。建模同时,各给药组大鼠分别灌胃给予维生素E、高剂量左归饮、低剂量左归饮及六味地黄丸药液进行干预,对照组与模型组分别给予等体积的蒸馏水。连续给药8周后,取大鼠肝、肾组织固定,利用HE染色法制作病理切片,进行显微观察;同时,采用免疫组化方法检测肝、肾组织中细胞凋亡相关蛋白(Bax、Bcl-2、Caspase-3)阳性表达水平,统计比较IOD值。结果 (1)与对照组比较,模型组大鼠肝、肾组织病理性损伤明显;与模型组比较,左归饮能够减轻D-半乳糖致大鼠衰老过程中出现的肝、肾组织病理性损伤。(2)与对照组比较,模型组大鼠肝组织中Bax和Caspase-3的IOD值均显著升高(P<0.01),Bcl-2的IOD值显著降低(P<0.01);与模型组比较,左归饮高剂量组大鼠肝组织中Bax、Caspase-3的IOD值均显著降低(P<0.01),Bcl-2的IOD值显著升高(P<0.01)。左归饮低剂量组大鼠肝组织中Bax和Caspase-3的IOD值明显降低(P<0.05),Bcl-2的IOD值略有升高,但差异无统计学意义(P>0.05)。(3)与对照组比较,模型组的肾组织中Bax和Caspase-3的IOD值均显著升高(P<0.01),Bcl-2的IOD值显著降低(P<0.01);与模型组比较,左归饮高剂量组大鼠肾组织中Bax、Caspase-3的IOD值均明显降低(P<0.01),Bcl-2的IOD值明显升高(P<0.01)。左归饮低剂量组大鼠肾组织中Bax的IOD值明显降低(P<0.05),但Bcl-2及Caspase-3的IOD值差异无统计学意义(P>0.05)。结论左归饮对肝、肾组织具有一定的保护作用,其作用机制可能是通过与影响肝、肾组织中细胞凋亡相关蛋白表达有关。 Objective To study the effects of Zuoguiyin on the morphological changes of liver and kidney and the expression of apoptosis related proteins in D-galactose induced subacute aging rat model and to explore the mechanism of its anti-aging effect. Methods Totally 72 healthy SD rats were randomly divided into 6 groups: control group, model group and administration group (vitamin E group, Zuoguiyin high and low dose group and Liuweidihuangwan group). In addition to 0.9% Na Cl solution subcutaneously injected into the neck and back of the control group, subcutaneous A-Aging models were established by subcutaneous injection of D-galactose in the back of the neck. At the same time, rats in each administration group were administered with vitamin E, high dose of Zuoguiyin, low dose of Zuoguiyin and Liuweidihuangwan respectively, and the control group and model group were given equal volume of distilled water respectively. After continuous administration for 8 weeks, the liver and kidney tissues of rats were fixed, and pathological sections were made by HE staining and observed microscopically. At the same time, immunohistochemistry was used to detect the expression of Bax, Bcl-2, Caspase-3) positive expression levels, statistical comparison of IOD values. Results (1) Compared with the control group, pathological damage of liver and kidney in the model group was obvious. Compared with the model group, Zuogui Yin could reduce pathological changes of liver and kidney in aging rats induced by D-galactose damage. (2) Compared with the control group, the IOD values ​​of Bax and Caspase-3 in the model group were significantly increased (P <0.01), the IOD values ​​of Bcl-2 significantly decreased (P <0.01) Compared with the high-dose Zuoguiyin group, the IOD values ​​of Bax and Caspase-3 were significantly decreased (P <0.01) and the IOD values ​​of Bcl-2 significantly increased (P <0.01). The IOD values ​​of Bax and Caspase-3 in liver tissue of Zuoguiyin low-dose group were significantly decreased (P <0.05), and the IOD values ​​of Bcl-2 slightly increased, but the difference was not statistically significant (P> 0.05). (3) Compared with the control group, the IOD values ​​of Bax and Caspase-3 in the model group were significantly increased (P <0.01) and the IOD values ​​of Bcl-2 significantly decreased (P <0.01) (P <0.01). The IOD of Bcl-2 was significantly increased (P <0.01). The IOD of Bax in the Zuoguiyin low dose group was significantly decreased (P <0.05), but the IOD values ​​of Bcl-2 and Caspase-3 were not significantly different (P> 0.05). Conclusion Zuoguiyin has a protective effect on liver and kidney tissues, and its mechanism may be related to the expression of apoptosis-related proteins in liver and kidney tissues.
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