补肾活血法对多囊卵巢综合征大鼠的防治作用及机制初步研究

目的观察补肾活血法治疗多囊卵巢综合征(PCOS)的疗效并探讨其作用机制。方法60只大鼠分为正常对照组、模型组、补肾活血法高、中、低剂量及罗格列酮组,每组10只。除正常对照组外,其余各组均制备PCOS模型。除正常对照组采用正常饮水外,其余大鼠造模结束后每日仍用5%葡萄糖溶液替代日常饮水。补肾活血方高、中、低剂量组通过人与大鼠体表面积比率换算法计算出灌胃给药量分别为68.66 g/kg,34.33 g/kg,17.17 g/kg。正常对照组和模型组大鼠每日按照体质量灌胃蒸馏水1 ml/100 g,1次/d,连续28 d;罗格列酮组灌胃罗格列酮3 mg/(kg·d),1次/d,连续28 d。检测各组大鼠体质量、卵巢重量和子宫重量,血清激素(LH、FSH和T)、炎症因子(HMGB1和TNF-α)、基因和蛋白(HMGB1、TLR4、NF-KBP65)的表达水平变化。结果与正常对照组比较,模型组、低剂量组和中剂量组的体质量、卵巢质量显著增加(P<0.05,P<0.01);模型组、补肾活血法低剂量组和中剂量组体质量、卵巢质量高于高剂量组和罗格列酮组,但组间比较差异无统计学意义(P>0.05);而模型组、低剂量组和中剂量组子宫质量显著低于正常对照组(P<0.05)。与正常对照组比较,各组LH、FSH和T含量均显著增加(P<0.05,P<0.01);模型组LH、FSH和T含量均高于高剂量组和罗格列酮组(P<0.05,P<0.01);高剂量组和罗格列酮组LH、FSH和T含量显著低于补肾活血法低、中剂量组(P<0.05)。与正常对照组比较,其余各组HMGB1和TNF-α含量均显著增加(P<0.05);模型组HMGB1和TNF-a含量均高于高剂量组和罗格列酮组(P<0.05);高剂量组和罗格列酮组HMGB1和TNF-α含量显著低于低、中剂量组(P<0.05)。与正常对照组比较,其余各组HMGB1、TLR4和NF-κB-P65基因的表达水平均显著增加(P<0.05);模型组HMGB1、TLR4和NF-κB-P65基因表达水平均高于高剂量组和罗格列酮组(P<0.05);补肾活血法高剂量组和罗格列酮组HMGB1、TLR4和NF-κB-P65基因表达水平显著低于低、中剂量组(P<0.01)。结论补肾活血法具有显著的降低PCOS大鼠体质量、卵巢质量、激素水平和炎症因子的作用,且对卵巢组织功能损伤具有保护作用,其作用机制可能与HMGB1/TLR4/NF-k BP65直接相关。

Preventive effect of bushen huoxue therapy on polycystic ovarian syndrome in the rats and the preliminary study on its effect mechanism

Objective To observe the effect and mechanism on polycystic ovary syndrome(PCOS) treated with bushen huoxue therapy(tonifying kidney and activating blood circulation. Methods A total of 60 rats were divided into a normal control group,a model group,the bushen huoxue groups of high dose,middle dose and low dose and a rosiglitazone group,10 rats in each one. Except the normal control group,PCOS models were prepared in the rest groups. The normal drink and feeding was used in the normal control group. In the rest groups,after end of modeling,5% glucose solution was used to replace daily water drinking. In the bushen huoxue groups of high dose,middle dose and low dose,according to the body surface area,the corresponding dose of intragastric administration were 68. 66 g/kg,34. 33 g/kg and 17. 17 g/kg successively. In the normal control group and the model group,according to the body weight,the distilled water,1 ml/100 g was used in intragastric administration,consecutively for 28 days. In the rosiglitazone group,rosiglitazone,3 mg(kg·d) was for intragastric administration,once a day,consecutively for 28 days. The body weight,ovary weight,uterine weight,and the expressions of serum hormones(LH,FSH and T),inflammatory factors(HMGB1 and TNF-α),genes and proteins(HMGB1,TLR4 and NF-KBP65) were determined in the rats of each group. Results Compared with the normal control group,in the low-dose and the middle-dose groups with herbal medication and the model group,the body weight and ovary weight were increased significant(P < 0. 05). In the model group and the low-dose and middle-dose groups of herbal medicine,the body weight and ovary weight were higher than the high-dose group of herbal medicine and the rosiglitazone group,and the differences were not significant among groups(P > 0. 05). In the model group,the low-dose and the middle-dose groups,the uterine weight was lower significantly than the normal control group(P < 0. 05). Compared with the normal control group,the levels of LH,FSH and T were increased significantly(P < 0. 05) in the other groups and the levels of LH,FSH and T in the model group were higher than the rest other groups(P < 0. 05,P < 0. 01). Compared with the normal control group,HMGB1 and TNF-α in the other groups were increased significantly(P < 0. 05). In the model group,the levels of HMGB1 and TNF-α were higher than the rest other groups(P < 0. 05,P < 0. 01). In the high-dose group of herbal medicine and the rosiglitazone group,they were lower significantly than the low-dose and the middle-dose groups(P < 0. 05). Compared with the normal control group,the expressions of HMGB1,TLR4 and NF-KB-P65 in the rest other groups were increased significantly(P < 0. 05). The expressions of HMGB1,TLR4 and NF-KB-P65 in the high-dose group of herbal medicine and the rosiglitazone group were lower significantly than the low-dose and the middle-dose group(P < 0. 01). Conclusion Berberine restores the estrous cycle and ovarian morphology in PCOS rats to a certain extent,and reduces the level of serum androgen,which may be one of the mechanisms of berberine in the treatment of PCOS.