补肾助孕方对黄体功能不全模型大鼠的治疗作用及机制

Effect and mechanism of Bushen Zhuyun Decoction in treatment of model rats with luteal phase defect

目的 探讨补肾助孕方对黄体功能不全(luteal phase defect,LPD)模型大鼠的治疗作用及机制。方法 将30只SD雌性大鼠随机分为空白、模型、中药低剂量、中药高剂量、阳性对照组。观察大鼠进入动情期后,上午除空白组外各组大鼠灌服米非司酮,空白组灌服生理盐水,下午各用药组灌服相应的药物,空白、模型组灌服生理盐水,连续3个动情周期,完成LPD的造模及给药后处死大鼠。检测大鼠血清雌二醇(E_(2))、孕酮(P)水平,KNDy神经元上3种神经肽及其受体(Kisspeptin/GPR54、NKB/NK3R、DYN/KOR)的mRNA与蛋白水平。结果 与空白组比较,模型组E_(2)、P均有下降趋势,但差异无统计学意义(P>0.05);与模型组相比,中药低、高剂量组E_(2)、P均有上升趋势,其中中药低、高剂量组E_(2)差异均有统计学(P<0.05)。与空白组比较,模型组KNDy神经元相关肽的mRNA水平均有下降趋势,其中GPR54、NK3R、DYN差异有统计学意义(P<0.05);与模型组比较,中药低剂量、高剂量组相关基因均有上升趋势,其中中药低、高剂量组GPR54、NK3R、DYN差异均有统计学意义(P<0.05)。与空白组比较,模型组KNDy神经元相关肽的蛋白表达均有下降趋势,其中NKB、DYN、KOR差异有统计学意义(P<0.05);与模型组相比,中药低剂量、高剂量组KNDy神经元相关蛋白均有上升趋势,其中中药高剂量组GPR54、NKB、NK3R、DYN、KOR差异有统计学意义(P<0.05)。结论 补肾助孕方可通过上调大鼠KNDy神经元相关神经肽的表达,改善卵巢黄体功能,对LPD起到治疗作用。

Objective To explore the therapeutic effect and mechanism of Bushen Zhuyun Decoction(BSZY-D) in the treatment of model rats with luteal phase defect(LPD).Methods A total of thirty female SD rats were randomly divided into control group,model group,BSZY-D(0. 9 g/100 g) group,BSZY-D(1. 8 g/100 g) group and dydrogesterone group. After the rats entered the estrous period,all groups were given mifepristone in the morning except the control group,and normal saline in the control group,corresponding drugs in the afternoon,and normal saline in the control and model groups. The rats were killed after LPD modeling and administration for 3 consecutive estrous cycles. The levels of serum estradiol(E_(2)) and progesterone(P),and the mRNA as well as the protein levels of the three neuropeptides and their receptors(Kisspeptin/GPR54,NKB/NK3R and DYN/KOR) were detected.Results Compared with the control group,the levels of serum E_(2)and P in the model group were decreased,but the difference was not statistically significant(P>0. 05);compared with the model group,the levels of E_(2) and P in BSZY-D(0. 9g/100g) group and BSZY-D(1. 8 g/100 g) group were increased,and there was significant difference in BSZY-D(0. 9 g/100 g) group,BSZY-D(1. 8 g/100 g) group with E_(2)(P<0. 05). The mRNA levels of KNDy neurons related peptide in the model group were decreased compared with the control group,the difference of GPR54,NK3R and DYN were statistically significant(P<0. 05);compared with the model group,the related genes in BSZY-D(0. 9 g/100 g) group and BSZY-D(1. 8 g/100 g) group showed an upward trend,and there was significant difference in GPR54,NK3R,and DYN in BSZY-D(0. 9 g/100 g) group and BSZY-D(1. 8 g/100 g)group(P<0. 05). The protein expression of KNDy neurons related peptide in model group was decreased compared with control group,and the differences of NKB,DYN and KOR were statistically significant(P<0. 05);compared with the model group,the KNDy neuron related protein in BSZY-D(0. 9 g/100 g) group and BSZY-D(1. 8 g/100 g) group showed an upward trend,and there was significant difference in GPR54, NKB, NK3R, DYN and KOR in BSZY-D(1. 8 g/100 g) group(P<0. 05).Conclusion Bushen Zhuyun Decoction might improve ovarian luteal endocrine by up regulating the expression of KNDy neurons related neuropeptide in rats,so as to have a therapeutic effect on LPD.