右归饮对轻微中枢神经损伤模型大鼠海马组织细胞自噬表达的影响

Effect of Yougui Yin on autophagy in hippocampus of rats with mild central nervous injury

目的观察右归饮对轻微中枢神经损伤模型大鼠海马组织细胞自噬相关基因和蛋白的表达,及海马组织细胞形态改变的影响。方法将48只SD雄性大鼠随机分为损伤模型组、右归饮组和空白对照组,每组16只。除空白组大鼠外,其余大鼠采用改良重物自由落体法进行轻微中枢神经损伤造模,造模结束后,右归饮组予右归饮(10g/kg体质量)进行灌胃,共持续4周。损伤模型组和空白对照组大鼠同时予等量的生理盐水灌胃。4周后处死所有大鼠,取出大鼠大脑海马组织,采用HE染色观察海马组织细胞形态学改变,以及免疫组化、QPCR检测自噬因子Beclin-1、LC3、mTOR的蛋白和mRNA表达量。结果 HE染色发现损伤模型组大鼠海马组织神经细胞数目稀少,排列紊乱,细胞边缘较为模糊,右归饮组大鼠海马组织细胞数目较损伤模型组多,排列较均匀,细胞边缘欠清晰。右归饮组大鼠自噬蛋白表达量与损伤模型组比较,差异有统计学意义[Beclin-1:(0.11±0.04)比(0.05±0.05),P<0.01;LC3:(0.03±0.02)比(0.01±0.00),P<0.01;mTOR:(0.03±0.03)比(0.09±0.04),P<0.05];右归饮组大鼠自噬相关基因Beclin-1与LC3 mRNA表达量与损伤模型组比较也显著上调[Beclin-1:(1.06±0.21)比(0.34±0.11),P<0.01;LC3:(1.75±0.21)比(1.12±0.09),P<0.01],mTOR表达下调[(0.91±0.17)比(1.74±0.73),P<0.01]。结论右归饮可以上调自噬的表达,减轻中枢神经细胞的破坏,缓解轻微中枢神经损伤的进展。

Objective To observe the effect of Youguiyin on the expression of autophagy-related genes and proteins in hippocampus of rats with mild central nervous system injury and the morphological changes of hippocampus cells. Methods Forty-eight Sprague Dawley(SD) male rats were randomly divided into injury model group, Youguiyin group and blank control group, 16 animals in each group. The rats in the injury model group and Yougui Yin group were modeled with slight central nervous system injury by the improved weight drop technique. After the modelling, the rats in Yougui Yin group were given Yougui Yin(10g/kg body weight) by gavage for 4 weeks. Rats in injury model group and blank control group were given the same amount of saline at the same time. Four weeks later, all rats were sacrificed and the hippocampal tissues were collected. The morphological changes of hippocampal cells were observed by HE staining, and the protein and mRNA expressions of autophagic factors Beclin-1, LC3 and mTOR were detected by immunohistochemistry and qPCR. Results HE staining showed that the number of hippocampal neurons in the injured model group was sparse, disordered and the cell edge was blurred. The number of hippocampal neurons in the Youguiyin group was more than that in the injured model group, and the arrangement was more uniform and the cell edge was not clear. The expression of autophagy protein in Youguiyin group was significantly higher than that in injury model group, Beclin-1:(0.11±0.04) vs (0.05±0.05), P<0.01;LC3:(0.03±0.02) vs (0.01±0.00), P<0.01;mTOR:(0.03±0.03) vs (0.09±0.04), P<0.05. Compared to the injury model group, the expression of autophagy-related gene Beclin-1 and LC3 in Youguiyin group was significantly increased, Beclin-1:(1.06±0.21) vs (0.34±0.11), LC3:(1.75±0.21) vs (1.12±0.09), mTOR:(0.91±0.17) vs (1.74±0.73), P<0.01, respectively. Conclusion Youguiyin can up-regulate the expression of autophagy, alleviate the damage of central nervous cells, and relieve the progress of slight central nervous injury.