艾灸足三里、太溪对肾虚-黄体抑制流产模型孕鼠蜕膜组织细胞凋亡及相关蛋白表达的影响

Effect of moxibustion of zúsānlǐ and tàixī on cell apoptosis and expression of related proteins in decidua tissues of pregnant rats with kidney deficiency-corpus luteum inhibition of abortion model

目的 观察艾灸足三里、太溪对肾虚-黄体抑制流产模型孕鼠蜕膜组织细胞凋亡及相关蛋白表达的影响。方法 选取32只雌性SD孕鼠(SPF级,8~10周,220~250 g),按随机数字表法分为正常组、模型组、地屈孕酮组和艾灸组,每组8只。正常组予生理盐水灌胃,其余组别均采用“羟基脲+米非司酮”制备SD孕鼠肾虚-黄体抑制流产模型,造模同时,地屈孕酮组予生理盐水配制地屈孕酮片3.02 mg/kg,2 ml/d灌胃;艾灸组予艾灸双侧足三里和太溪穴,每天15 min(/次·穴),连续干预9 d后取材。计算各组宫内胚胎数并测量子宫直径,TUNEL法检测各组大鼠蜕膜组织细胞凋亡并计算凋亡指数(AI)值,免疫组织化学测定各组大鼠蜕膜组织Bax、Bcl-2和Caspase-3蛋白表达。结果 与正常组比较,模型组大鼠子宫直径减小,差异有统计学意义(P <0.05);与模型组比较,艾灸组子宫直径增大,差异有统计学意义(P <0.05);正常组和模型组,模型组和艾灸组宫内胚胎总数比较,差异无统计学意义(P> 0.05)。与正常组比较,模型组Bax、Caspase-3蛋白平均OD值升高,差异有统计学意义(P <0.05),与模型组比较,艾灸组Bax、Caspase-3蛋白平均OD值降低,差异有统计学意义(P <0.05)。与正常组比较,模型组Bcl-2蛋白平均OD值降低,差异有统计学意义(P <0.05),与模型组比较,艾灸组Bcl-2蛋白平均OD值升高,差异有统计学意义(P <0.05)。与正常组比较,模型组细胞AI值升高,差异有统计学意义(P <0.05),与模型组比较,艾灸组细胞AI值降低,差异有统计学意义(P <0.05)。结论 艾灸足三里和太溪穴可能通过上调肾虚-黄体抑制流产孕鼠蜕膜组织Bcl-2蛋白表达并下调其Bax、Caspase-3蛋白过量表达,降低其AI,以调控母胎界面细胞凋亡。

Objective To observe the effect of moxibustion of zúsānlǐ and tàixī on cell apoptosis and expression of related proteins in decidua tissues of pregnant rats with kidney deficiency-corpus luteum inhibition of abortion model. Methods Thirty-two pregnant female SD rats(SPF, 8-10 weeks, 220-250 g) were divided into normal group, model group,Dydrogesterone group, and moxibustion group according to the random number table method, with eight rats in each group.The normal group was given normal saline intragastric administration, and the other groups were given “Hydroxyurea +Mifepristone”to prepare the kidney deficiency-corpus luteum inhibition of abortion model of pregnant SD rats. At the same time, the Dydrogesterone group was given Dydrogesterone Tablets 3.02 mg/kg, 2 ml/day by gavage. In the moxibustion group,bilateral zúsānlǐ and tàixī points were moxibusted for 15 min/(time·point) every day, and the samples were collected after nine days of continuous intervention. The number of embryos in each group was calculated and the diameter of uterus was measured. The cell apoptosis of decidua tissues of each group was detected by TUNEL method, and the apoptosis index(AI) value was calculated. The protein expressions of Bax,Bcl-2, and Caspase-3 in decidua tissues of each group were determined by immunohistochemistry. Results Compared with the normal group, the uterine diameter of the model group was decreased, and the difference was statistically significant(P < 0.05). Compared with the model group, the uterine diameter of the moxibustion group was increased, and the difference was statistically significant(P < 0.05). There were no significant differences in the total number of intrauterine embryos between normal group and model group, model group and moxibustion group(P > 0.05). Compared with the normal group, the average OD value of Bax and Caspase-3 protein in the model group was increased, and the differences were statistically significant(P < 0.05).Compared with the model group, the average OD value of Bax and Caspase-3 protein in the moxibustion group was decreased, and the differences were statistically significant(P < 0.05). Compared with the normal group, the average OD value of Bcl-2 protein in the model group was decreased, and the difference was statistically significant(P < 0.05).Compared with the model group, the average OD value of Bcl-2 protein in the moxibustion group was increased, and the difference was statistically significant(P < 0.05). Compared with the normal group, the cell AI value of the model group was increased, and the difference was statistically significant(P < 0.05). Compared with the model group, the cell AI value of the moxibustion group was decreased, and the difference was statistically significant(P < 0.05). Conclusion Moxibustion at zúsānlǐ and tàixī may regulate cell apoptosis at the maternal-fetal interface by up-regulating the expression of Bcl-2protein, down-regulating the overexpression of Bax and Caspase-3 protein and reducing the AI in decidua tissues of pregnant rats with kidney deficiency and corpus luteum inhibition of abortion.