补肾更年方对围绝经期综合征大鼠神经-内分泌-免疫网络功能的影响

Effect and Mechanism of Bushen Gengnian Formula on the Function of Neuroendocrine Immune Network in Rats with Perimenopausal Syndrome

目的通过观察补肾更年方对围绝经期综合征大鼠行为学指标、胸腺、子宫、肾上腺指数、血清和下丘脑NE、5-HT含量和子宫HSP70、ERα36蛋白表达的影响,探讨其对围绝经期综合征(perimenopausal syndrome,PMS)大鼠神经-内分泌-免疫网络功能的效应机制。方法采用摘除卵巢法复制PMS大鼠模型,将连续5 d动情周期紊乱的大鼠随机分为模型组、西药组、补肾更年方大、中、小剂量组,每组10只,10只为对照组大鼠,西药组采用1 mL/100 g剂量的己烯雌酚水溶液灌胃,补肾更年方三种剂量组分别以0.015 mL·g^(-1)、0.010 mL·g^(-1)、0.005 mL·g^(-1)剂量的水煎液灌胃,对照组和模型组以1 mL/100 g的生理盐水灌胃,连续灌胃45 d,干预结束后,观察各组大鼠一般情况,采用旷场实验和高架十字迷宫实验测试大鼠行为学改变,计算各组大鼠胸腺、子宫、肾上腺指数,采用放免法和化学发光法分别检测各组大鼠血清、下丘脑NE、5-HT含量,采用western blot法检测各组大鼠子宫HSP70和ERα36蛋白表达。结果与模型组比较,西药和补肾更年方三个剂量干预均可改善大鼠一般情况;与模型组比较,西药和补肾更年方大、中剂量干预可明显增加大鼠中央区域停留时间、总移动距离、开臂停留时间和开臂进入次数(P<0.05),升高大鼠胸腺、子宫、肾上腺指数(P<0.05),降低大鼠血清、下丘脑NE、5-HT含量(P<0.05),上调大鼠子宫HSP70和ERα36蛋白表达(P<0.05)。结论补肾更年方可能通过提高PMS大鼠胸腺、子宫、肾上腺指数,降低其血清、下丘脑NE、5-HT含量,上调其子宫HSP70和ERα36蛋白表达,综合调控PMS大鼠神经-内分泌-免疫网络功能,改善焦虑样行为,发挥对PMS的治疗效应。

Objective To investigate the effects of the Bushen Gengnian formula on behavioral indicators,thymus,uterus,adrenal index,serum and hypothalamic NE and 5-HT content,and uterine HSP70 and ERα36 protein expression in rats with perimenopausal syndrome,and to explore its mechanism of action on the neuroendocrine immune network function of PMS rats.Methods The PMS rat model was replicated by ovariectomy.Rats with disrupted estrous cycles for 5 consecutive days were randomly divided into a model group,a western medicine group,and a large,medium,and small dose group of Bushen Gengnian formula,with 10 rats in each group.The control group consisted of 10 rats.The western medicine group was given a 1 mL/100 g dose of diethylstilbestrol aqueous solution by gavage,while the Bushen Gengnian formula three dose groups were given 0.015 mL·g^(-1)、0.010 mL·g^(-1)、0.005 mL·g^(-1) doses of decoction by gavage,respectively.The control group and model group were given 1 mL/100g of physiological saline by gavage for 45 consecutive days.After the intervention,the general condition of each group of rats was observed,and an open field experiment was conducted.The elevated cross maze experiment was used to test the behavioral changes of rats,and the thymus,uterus,and adrenal indices of each group of rats were calculated.The serum,hypothalamic NE,and 5-HT contents of each group of rats were detected by radioimmunoassay and chemiluminescence,respectively,Western blot was used to detect the expression of HSP70 and ERα36 proteins in the uterus of rats in each group.Results Compared with the model group,the three doses of Western medicine and Bushen Gengnian formula intervention can improve the general condition of rats;Compared with the model group,Western medicine and the Bushen Gengnian formula at high and medium doses significantly increased the central area residence time,total movement distance,arm opening residence time,and number of arm opening entries in rats(P<0.05),increased thymus,uterus,and adrenal indices(P<0.05),decreased serum and hypothalamic NE and 5-HT levels in rats(P<0.05),and upregulated HSP70 and ERα36 protein expression in rats uterus(P<0.05).Conclusion The Bushen Gengnian formula may enhance the thymus,uterus,and adrenal indices of PMS rats,reduce their serum and hypothalamic NE and 5-HT levels,upregulate their uterine HSP70 and ERα36 protein expression,comprehensively regulate the neuroendocrine immune network function of PMS rats,improve anxiety like behavior,and exert therapeutic effects on PMS.