补肾疏肝方联合五行音乐对卵巢储备功能减退大鼠的实验研究

Experimental study on Bushen Shugan Decoction combined with five elements music on rats with decreased ovarian reserve

关键词:  补肾疏肝方  五行音乐  大鼠  卵巢储备功能减退  实验研究

Key words:  Bushen Shugan Decoction  five elements music  rat  decreased ovarian reserve  experimental study

摘要:

[目的] 探讨补肾疏肝方联合五行音乐对卵巢储备功能减退大鼠的干预作用。[方法] 将54只有规律动情周期、雌性SD大鼠，随机分为空白组、模型组、补佳乐组、中药低剂量联合音乐组、中药中剂量联合音乐组、中药高剂量联合音乐组6组，每组9只。实验开始后，每日测定6组大鼠动情周期，对照组正常进食水，其余5组采用慢性不可预见性应激法联合雷公藤多苷片灌胃建立卵巢储备功能减退模型；模型成功后，对照组、模型组正常进食水，补佳乐组给予西药补佳乐灌胃，中药低、中、高剂量联合音乐组3组分别给予补肾疏肝药物，灌胃同时每日听音乐45 min，连续21 d。于实验第42天，进行糖水实验、旷场实验，麻醉，腹主动脉取血，处死后取卵巢并称质量计算卵巢指数，采用酶联免疫吸附（ELISA）测定血中促卵泡生成素（FSH）、促黄体生成素（LH）、雌二醇（E2），对卵巢组织进行苏木精-伊红（HE）染色，观察各级卵泡及颗粒细胞的情况。[结果] 实验结束后，模型组大鼠体质量增加缓慢，被毛稀松，饮食、饮水均减少。与模型组相比，其余各组上述表现有所改善。造模结束后，与空白组比较，其余5组大鼠动情次数减少、糖水偏嗜度下降、直立及垂直次数减少，E2升高，FSH及LH均下降，差异有统计学意义（P<0.05），提示造模成功。治疗后与模型组比较，中药低、中、高剂量联合音乐组与补佳乐组E2均升高，FSH与LH均降低，差异有统计学意义（P<0.05）。治疗前后组内比较，治疗后补佳乐组、中药高剂量联合音乐组大鼠动情次数增加、糖水偏嗜度升高、水平及直立次数增多，差异有统计学意义（P<0.05），其余各组大鼠差异无统计学意义（P>0.05），但中药低剂量联合音乐组大鼠在治疗前后动情次数均增加、糖水偏嗜度均增高、水平及直立次数均增加，提示药物干预对促进模型的恢复有一定的作用。卵巢病理切片结果：与模型组比较，其余各组卵巢病理切片显示卵泡形态较好，颗粒细胞分级增多，尤其是补佳乐组和中药高剂量联合音乐组。[结论] 补肾疏肝方联合五行音乐对卵巢储备功能减退大鼠的恢复有一定的促进作用。

Abstract:

[Objective] To investigate the intervention effect of Bushen Shugan Decoction combined with five elements music on rats with decreased ovarian reserve.[Methods] Fifty-four female SD rats with regular estrous cycle were randomly divided into 6 groups,including blank group,model group,bujiale group,low-dose Chinese medicine combined music group,middle-dose Chinese medicine combined music group and high-dose Chinese medicine combined music group,with 9 rats in each group. After the start of the experiment,the estrus cycle of the rats in 6 groups was measured every day,the control group was fed with water normally,the other 5 groups were given chronic unpredictable stress combined with tripterygium wilfordii polyglycoside tablets by gavage to establish the model of decreased ovarian reserve. After the model was successful,the control group and the model group were fed with water normally,the Western medicine group was given bujiale by gavage,and the 3 groups of Chinese medicine low,middle and high dose combined with music were given respectively low,middle and high doses of Bushen Shugan Decoction by gavage,while listening to music for 45 minutes a day for 21 days. On the 42nd day of the experiment,sugar water test,open field test,anesthesia,abdominal aorta blood samples were taken,and the ovaries were taken and weighed to calculate the ovarian index. The levels of follicle stimulating hormone (FSH),luteinizing hormone (LH) and estradiol (E2) in blood were determined by enzyme linked immunosorbent assay (ELISA). The ovarian tissues were stained with HE to observe the follicular and granulosa cells at all levels.[Results] After the experiment,the rats in the model group gained weight slowly,their coats were loose,and their diet and drinking water decreased. Compared with the model group,the above-mentioned performances of the other groups have improved. After modeling,compared with the blank group,the number of estrus,sugar water preference,level and standing times decreased,E2 increased,FSH and LH decreased in the other five groups,and the difference was statistically significant (P<0.05),indicating that modeling was successful. After treatment,compared with the model group,E2 in the low,middle and high dose Chinese medicine combined music group and bujiale group increased,FSH and LH decreased,and the difference was statistically significant (P<0.05). Compared before and after treatment,the estrus times,sugar water preference,level and standing times of rats in bujiale group and high-dose Chinese medicine combined music group increased significantly (P<0.05),but there was no significant difference among other groups (P>0.05). However,the estrus times,sugar water partiality,level and standing times of rats in low-dose Chinese medicine combined music group increased after treatment,suggesting that drug intervention has a certain effect on promoting the recovery of the model. Ovarian pathological section results:Compared with the model group,ovarian pathological sections of other groups showed better follicular morphology and increased granulosa cell classification,especially bujiale group and high-dose Chinese medicine combined music group.[Conclusion] Bushen Shugan Decoction combined with five elements music can promote the recovery of ovarian reserve function in rats.