加味补肝汤对糖尿病大鼠背根节细胞凋亡的影响

Effects of Jiawei Bugan decoction on cell apoptosis of dorsal root ganglion in diabetic rats

目的观察加味补肝汤对糖尿病大鼠背根节细胞凋亡的影响,探讨其神经保护作用机制。方法以链脲佐菌素(streptozotocin,STZ)诱发糖尿病模型。分别给药后4、8周处死大鼠,应用化学比色法测定血清中丙二醛(MDA)和谷胱甘肽(GSH)含量;用免疫组织化学法和脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)法分别检测背根节细胞活化半胱天冬酶-3(Caspase-3)的蛋白表达以及细胞凋亡。结果(1)造模后4周、8周模型组大鼠血清中MDA水平明显高于同一时间点正常组,差异具有统计学意义(P<0.01),而GSH水平明显低于同一时间点正常组,差异具有统计学意义(P<0.01)。随病程的延长大鼠血清中MDA水平逐渐增加,而GSH水平逐渐降低。与同一时间点模型组相比,加味补肝汤能降低血清中MDA水平,提高GSH水平,差异具有统计学意义(均P<0.05);(2)在正常组大鼠背根节细胞中未见活化的Caspase-3的蛋白表达和细胞凋亡,模型组大鼠上述表达均增加,加味补肝汤干预后其表达均较同一时间点模型组表达明显减少,差异具有统计学意义(P<0.05)。结论加味补肝汤抑制Caspase-3的活化,减少细胞凋亡,可能与调节机体内的氧化应激状态有关,具有良好的抗氧化作用,从而达到神经保护作用。

Objective To observe the effects of Jiawei Bugan decoction （JWBG） on cell apoptosis of dorsal root ganglion（DRG）in diabetic rat and study the mechanism of the decoction on nervous protective effect. Methods The diabetic rats models were induced by streptozotocin, and killed in 4th and 8th week. The content of MDA and GSH in serum were detected with chromatometry; Caspase-3 expression of protein in DGR with immunohistochemical and apoptosis of DGR with the method of TUNEL. Results The serum MDA in model group was significantly increased on the 4th and 8th weeks than that in the control group after induced-model（P〈0.01）, but the content of GSH in serum were obviously lower than that in the control group in the corresponding time period （P〈0.01）. Content of MDA in serum were increased gradually While the level of GSH decreased gradually along with extension of disease course. Compared with the model group, the decoction could decrease the content of serum MDA and increase the level of HSH （P〈0.05）. The protein expression of activated caspase-3 and apoptosis of cell were not found in the control group, but the expression in JWBG group were increased in the model group; the expression of caspase-3 and apoptosis of DRG were decreased significantly based on the action of JWBG decoction （P〈0.05）. Conclusion JWBG decoction can inhibit the transcription of caspase-3 mRNA and activation of caspase-3, and has a good antioxidant effect, which may be related to regulating the oxidative stress state of organism.