补肾化痰复方对多囊卵巢综合征大鼠的作用机制研究

Effect of Bushen-Huatan compound on polycystic ovary syndrome in rats

目的观察补肾化痰复方通过调控胰岛素分泌和卵巢细胞增殖能力从而影响多囊卵巢综合征(PCOS)大鼠的作用,探究补肾化痰复方对减缓PCOS疾病的发生是否具有积极作用。方法经阴道涂片筛选出80只动情周期正常的雌性大鼠,采用随机数字表法将大鼠分为Ctrl组、PCOS组、低剂量组、高剂量组,每组20只。PCOS组、低剂量组、高剂量组建立PCOS大鼠模型,低剂量组、高剂量组分别给药补肾化痰复方60、120 mg/kg,PCOS组、Ctrl组采用同种方式加入等体积蒸馏水。实时监测四组大鼠游离脂肪酸、黄体酮、总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)水平;通过监测四组大鼠禁食180 min血糖变化检测其胰岛素糖耐量试验(ITT)值;采用CCK8检测大鼠卵巢颗粒细胞增殖情况。比较四组大鼠游离脂肪酸、黄体酮、TC、TG、LDL-C和HDL-C水平;禁食不同时间后ITT值;不同剂量补肾化痰复方组大鼠卵巢颗粒细胞增殖情况。结果PCOS组大鼠黄体酮、HDL-C水平显著低于Ctrl组,游离脂肪酸、LDL-C、TC、TG水平高于Ctrl组,差异具有统计学意义(P<0.05)。低剂量组黄体酮高于PCOS组,游离脂肪酸、LDL-C低于PCOS组,差异具有统计学意义(P<0.05);低剂量组TC、HDL-C和TG水平与PCOS组比较差异无统计学意义(P>0.05)。高剂量组黄体酮、HDL-C水平高于PCOS组,游离脂肪酸、LDL-C、TC、TG水平低于PCOS组,差异具有统计学意义(P<0.05)。在相同时间点,PCOS组大鼠ITT值均高于Ctrl组,差异具有统计学意义(P<0.05);高、低剂量组ITT值与Ctrl组比较差异均无统计学意义(P>0.05);且高剂量组的ITT值随时间的变化幅度相较于低剂量组而言更小。0μM组、10μM组、50μM组和100μM组大鼠卵巢颗粒细胞相对活性分别为(91.00±11.36)%、(63.33±8.74)%、(53.00±11.79)%和(41.00±14.11)%,比较差异具有统计学意义(P<0.05);10μM组、50μM组和100μM组大鼠卵巢颗粒细胞相对活性明显低于0μM组,差异具有统计学意义(P<0.05),并且随着剂量梯度升高,下降越显著。结论补肾化痰复方能够显著升高PCOS大鼠黄体酮和HDL-C水平,下调游离脂肪酸、LDL-C、TC、TG水平,不仅会较大程度地改善胰岛素耐量,还有降低大鼠卵巢颗粒细胞的生理增殖活跃的作用。

Objective To observe the effect of Bushen Huatan compound on polycystic ovary syndrome(PCOS)rats by regulating insulin secretion and ovarian cell proliferation,and explore whether Bushen Huatan compound has a positive effect on reducing the occurrence of PCOS disease.Methods 80 female rats with normal estrous cycle were screened by vaginal smear,and the rats were divided into Ctrl group,PCOS group,low-dose group,and high-dose group according to random numerical table,with 20 rats in each group.The PCOS rat model was established in PCOS group,low-dose group and high-dose group,and the low-dose group and high-dose group were administered with 60 and 120 mg/kg of Bushen Huatan compound respectively,and equal volume of distilled water was added to PCOS group and Ctrl group in the same way.The levels of free fatty acid,progesterone,total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C)were monitored in real time;the insulin tolerance test(ITT)value was detected by monitoring the blood glucose changes of rats in the four groups after fasting for 180 min;CCK8 was used to detect the proliferation of granulosa cells in rats.Comparison was made on levels of free fatty acid,progesterone,TC,TG,LDL-C and HDL-C;ITT values after different times of fasting in the four groups;and the proliferation of granulosa cells in different doses of Bushen Huatan compound groups.Results The levels of progesterone and HDL-C in the PCOS group were significantly lower than those in the Ctrl group,and the levels of free fatty acids,LDL-C,TC,and TG were higher than those in the Ctrl group.All the differences were statistically significant(P<0.05).The progesterone in the low-dose group was higher than that in the PCOS group,and the free fatty acids and LDL-C were lower than those in the PCOS group.All the differences were statistically significant(P<0.05).There was no statistically significant difference in the levels of TC,HDL-C and TG between the low-dose group and the PCOS group(P>0.05).The levels of progesterone and HDL-C in the high-dose group were higher than those in the PCOS group,and the levels of free fatty acids,LDL-C,TC,and TG were lower than those in the PCOS group.All the differences were statistically significant(P<0.05).At the same time point,the ITT values of the rats in the PCOS group were higher than those in the Ctrl group,and the difference was statistically significant(P<0.05).The ITT value of the high-dose group and low-dose group was not statistically significant compared with the Ctrl group(P>0.05).The ITT value of the high-dose group changed less over time than the low-dose group.The relative viability of granulosa cells in 0μM group,10μM group,50μM group and 100μM group were(91.00±11.36)%,(63.33±8.74)%,(53.00±11.79)%and(41.00±14.11)%,and the differences were statistically significant(P<0.05).The relative viability of granulosa cells in the 10μM group,50μM group and 100μM group was significantly lower than that in the 0μM group,and the differences were statistically significant(P<0.05),and the decrease was more significant with the increase of the dose gradient.Conclusion Bushen Huatan compound can significantly increase progestin and HDL level,and decrease free fatty acid level,LDL-C,TC and TG level in PCOS rats.It can not only improve insulin tolerance to a great extent,but also decrease the physiological proliferation activity of granulosa cells in the ovary of rats.