药食合用对围绝经期综合征模型大鼠神经-内分泌-免疫网络细胞凋亡及ER表达的影响

Effects of Drug-Food Combination on Apoptosis and ER Expression of Neuroendocrine Immune Network in Rats with Perimenopausal Syndrome

目的观察更年滋肾汤合玉女酥对雌性去势大鼠下丘脑、垂体、子宫、胸腺、脾脏细胞凋亡及ER表达的影响,从神经-内分泌-免疫网络探讨药食合用治疗围绝经期综合征的作用机制。方法采取双侧卵巢切除法建立围绝经期综合征大鼠模型,2015年10月—2016年4月,选取上海斯莱克实验动物中心健康、12周龄、雌性未孕SD大鼠60只,随机分为正常对照组、模型组、中药(更年滋肾汤)组、食疗(玉女酥)组、药食合用组,每组12只。给药(食)6周后,处死大鼠取丘脑、垂体、子宫、胸腺、脾脏组织,检测如下指标:TUNEL方法检测细胞凋亡率;Western blot技术检测ERα蛋白的表达变化;Real time PCR方法检测ERαm RNA的表达变化。结果 (1)细胞凋亡率变化:下丘脑、垂体各组差异无统计学意义;模型组子宫(13.12±0.066)明显高于对照组(3.28±0.075),中药组(6.35±0.038)、药食合用组(4.90±0.034)明显低于模型组;模型组胸腺(3.46±0.215)明显高于对照组(1.70±0.045),中药组(2.51±0.038)、药食合用组(2.18±0.034)明显低于模型组;模型组脾脏(4.05±0.205)明显高于对照组,中药组(2.15±0.115)、药食合用组(1.64±0.082)明显低于模型组;差异有统计学意义(P<0.05,P<0.01)。(2)ERα蛋白表达变化:各组织模型组明显低于正常组(P<0.05);除胸腺外,其他组织中药组、药食合用组均明显高于模型组(P<0.05,P<0.01)。(3)ERαm RNA表达变化:模型组垂体(0.210±0.012)、下丘脑(0.197±0.015)、子宫(0.168±0.016)、脾脏(0.137±0.016)明显低于对照组(P<0.05);中药组垂体(0.458±0.026)、下丘脑(0.410±0.028)、子宫(0.367±0.037)、脾脏(0.301±0.025)高于模型组(P<0.05);食疗组子宫(0.349±0.021)、脾脏(0.282±0.018)高于模型组(P<0.05);药食合用组垂体(0.523±0.025)、下丘脑(0.488±0.024)、子宫(0.743±0.042)、脾脏(0.723±0.069)高于模型组(P<0.05)。结论药食合用(更年滋肾汤合玉女酥)可通过影响雌性去势大鼠下丘脑、垂体、子宫、胸腺、脾脏细胞凋亡及ER表达,调节神经-内分泌-免疫网络,从而改善围绝经期综合征。

Objective This paper tries to observe the effect of Gengnianzishen Decoction and Yunv crisp on the apoptosis and the expression of ER of hypothalamus, pituitary, uterus, thymus, spleen of ovariectomized female rat, and explore themechanism of drug-food combination in the treatment of peri menopausal syndrome from the neuroendocrine immune network. Methods The bilateral ovariectomy was used to establish the perimenopausal syndrome rat model. 60 health, 12 week old female SD rats from October 2015 to April 2016 were selected from Shanghai SLRC Animal Laboratory Center, and were randomly divided into the control group, model group, Chinese medicine（Gengnianzishen Decoction） group, diet therapy（Yunv crisp） group and drug-food combination group, with 12 cases in each group. After treating drugs（food） for continuous 6 weeks, the rats were killed and their hypothalamus, pituitary, uterus, thymus, spleen were taken out and used to detect the following indicators： the cell apoptosis rate was detected by TUNEL method; the expression of ER alpha protein by Western blot; expression of ER alpha m RNA by Real time PCR. Results（1）The apoptosis rate changes： there was no significant difference in hypothalamus and pituitary between the groups; the uterus of model group was（13.12±0.066） significantly higher than the control group of（3.28±0.075）, Chinese medicine group of（6.35±0.038）, drug-food combination group of（4.90±0.034）, significantly lower than that of model group; the thymus of the model group was（3.46 ±0.215）, significantly higher than the control group of（1.70±0.045）, Chinese medicine group of（2.51±0.038）, drug-food combination group of（2.18±0.034）, significantly higher than that of model group; the spleen of the model group was（4.05 ±0.205）, significantly higher than the control group, Chinese medicine group was（2.15±0.115）, drug-food combination group of（1.64±0.082）, significantly higher than that of model group（P0.01,P0.05）.（2）Expression of ER alpha protein： the model group decreased more significantly than the control group（P0.05）; except for the thymus, Chinese medicine group, food and drug combination group were increased significantly than the model group（P0.05, P0.01）.（3）expression of ER alpha m RNA： hypothalamus was（0.197 ±0.015）, pituitary was（0.210 ±0.012）, uterus was（0.168 ±0.016）, spleen was（0.137 ±0.016） of the model group, decreased significantly than the control group（P0.05）; hypothalamus was（0.410±0.028）, pituitary was（0.458±0.026）, uterus was（0.367±0.037）, spleen was（0.301±0.025） of the Chinese medicine group, increased significantly compared with the model group（P0.05）; The uterus（0.349±0.021） and spleen（0.282±0.018） were higher than the mooled grap（P0.05） hypothalamus was（0.488±0.024）, pituitary was（0.523 ±0.025）, uterus was（0.743±0.042）, spleen was（0.723 ±0.069） of the food and drug combination group, increased significantly compared with the model group（P0.05）. Conclusion The combination of medicine and food（Gengnianzishen Decoction and Yunv crisp） can affect the apoptosis and expression of ER in hypothalamus, pituitary, uterus, thymus, spleen of ovariectomized female rat, regulating network in rats with perimenopausal syndrome, so as to improve perimenopausal syndrome.