摘要
目的:探讨“补肾宁”对氢化可的松法复制肾阳虚排卵障碍模型大鼠,神经递质及生殖机能调控作用机制。
方法:成年雌性SD大鼠60只,随机分为6组,每组10只。第1组为空白对照组,后5组建立动物模型,分别为模型对照组、克罗米芬组、补肾宁低、中、高剂量组。空白对照组每日肌注等容积生理盐水,其余5组,每日下肢肌注氢化可的松注射液36mg/kg,连续11天,制备肾阳虚排卵障碍模型大鼠。11天后按组别分别用药治疗,治疗期为15天。实验观察模型大鼠用药前后体重及血清E2、FSH、LH激素水平的变化;大鼠下丘脑DA、NE、5-HT含量的变化;下丘脑、卵巢、子宫显微光镜下组织形态学的变化;卵巢、子宫超显微结构的改变;卵巢、子宫FSHR的表达;卵巢FSHRmRNA的表达,并进行组间的比较。
结果:①补肾宁各剂量组能够提高模型大鼠用药前后的体重,明显优于与模型组、克罗米芬组,相比较有显著性差异(P<0.01),补肾宁高剂量组优于补肾宁低剂量组(P<0.05)。②补肾宁各剂量组及克罗米芬组均能明显提高模型大鼠血清E2、FSH、LH水平以及调节下丘脑DA、NE、5-HT,其中克罗米芬组及补肾宁高剂量组作用最为显著。③模型对照组大鼠卵巢中生长卵泡及黄体数目明显减少,补肾宁各剂量组及克罗米芬组能显著增加不同阶段卵泡及黄体数目,其中补肾宁高、中剂量组、克罗米芬组与空白对照组相类似;补肾宁高、中剂量组能够明显改善模型大鼠子宫间质和腺体的形态,接近正常对照组,优于克罗米芬组;各组间下丘脑光镜观察无显著性差异。④补肾宁能减轻模型大鼠卵巢、子宫超微结构的损伤,其中高、中剂量组效果较为明显,优于克罗米芬组。⑤补肾宁各剂量组与克罗米芬组均能增强下丘脑的GnRH表达强度,与模型对照组比较有显著性差异(P<0.01),其中补肾宁高与克罗米芬组剂量组作用最为明显,与空白对照组无显著性差异(P>0.05)。⑥补肾宁组与克罗米芬组均能不同程度的增强卵巢、子宫的FSHR的表达强度,其中补肾宁高、中剂量组作用显著,与模型对照组比较有显著性差异(P<0.01),与空白对照组无显著性差异(P>0.05),其中补肾宁高作用最为明显,作用优于克罗米芬组(P<0.05)。⑦补肾宁组与克罗米芬组均能增强卵巢FSHRmRNA的表达强度,与模型对照组比较差异显著(P<0.05),其中补肾宁高、中剂量组作用最为明显,与模型对照组比较有极显著性差异(P<0.01),与空白对照组无显著性差异(P>0.05),补肾宁高剂量组卵巢FSHRmRNA的表达强度优于克罗米芬组(P<0.05)。
结论:补肾宁能够整体调节肾阳虚模型大鼠下丘脑-垂体-卵巢轴的机能。明显提高模型大鼠造模前后的体重及血清E2、FSH、LH水平,调节大鼠下丘脑DA、NE、5-HT含量的变化;改善模型大鼠子宫及卵巢的光镜组织学形态及子宫及卵巢超微结构;增强模型大鼠下丘脑GnRH以及卵巢、子宫FSHR的表达强度;增强模型大鼠卵巢FSHRmRNA的表达强度。补肾宁能够促进卵泡的发育与排出,恢复卵巢的功能,整体调节生殖轴功能,作用优于克罗米芬。
Objective:To observe the effect and mechanism which BuShenNing(BSN)decoction bushenning regulating rat neurotransmitters and reproductive function in the ovulation inhibition of kidney yang, which replicate by applying hydoreortisone.
Methods:The sixty mature female Sprague-Dawley rats were divided into six groups radomly:normal contrast group,model contrast group,clomifene group, BSN low,middle and high dosage group. normal contrast group used physiological saline, the other groups replicated kidney-yang defieiency model by muscle injecting hydoreortisone 36mg/kg,for 11 day.After 11 day,used the corresponding medicine treatment by 15 day.Observe each group the change of rat body weight ; the level of serum estrogen ( E2 ),Follicle-Stimulating Hormone( FSH) and Luteinizing Hormone( LH); the content of hypothalamus DA、NE、5-HT ,each group the hypothalamus, ovarian , uterine changed in histomorphological differences by light microscope; ultrastrucure changed of ovarian, uterine; the expression of hypothalamus Gondaotropin releasing hormone( GnRH); the expression of hypothalam, usterine, Follicle-Stimulating Hormon receptor( FSHR); the expression of Follicle-Stimulating Hormon receptor mRNA( FSHR mRNA).
Results:①After taken medicine,the weight of rat in BSN groups heighten evidently,have significant differerce compared with model contrast group and clomifene group( P<0.01),and superiord to BSN low dosage group(P<0.05).②BSN low,middle and high dosage group and clomifene group all increased level of the rat serum E2, FSH and LH, and regulate the contents of DA,NE and 5-HT,clomifene group and BSN high dosage group the most significant.③BSN low, middle and high dosage group improvd the structure damage of ovarian,uterine of deficient kidney yang rat,the higher the dose, the effect of the more obvious; model contrast group decreased in number of ovarian growth follicle and luteum,BSN low,middle,high dosage group and clomifene group could increase in quantity of ovarian growth follicle and luteum, BSN middle, high dosage group and clomifene group were similar to normal contrast group; BSN middle and high dosage group improved the morphology of rat uterine stromal and glandular , were superiored to clomifene group, were similar to normal contrast group;hypothalamus of each group were observed no significant difference by light microscope.④BSN low,middle and high dosage group improvd ultrastrucure damage of ovarian , uterine , BSN middle and high dosage group were superiored to clomifene group.⑤BSN low, middle, high dosage group and clomifene group all could enhance the express of hypothalamus GnRH, have significant differerce compared with model contrast group ( P<0.01 ),clomifene group and BSN high dosage group the most significant.⑥BSN low, middle, high dosage group and clomifene group all could enhance the express of ovarian and uterine FSHR, BSN middle and high dosage group were similar to normal contrast group ( P > 0.05 ), were superiored to clomifene group( P< 0.05)and model contrast group( P<0.01).⑦BSN low,middle,high dosage group and clomifene group all could enhance the express of ovarian FSHRmRNA,were superiored to model contrast group( P< 0.05),BSN middle and high dosage group were significant superiored to model contrast group( P<0.01),were similar to normal contrast group( P> 0.05), BSN high dosage group was superiored to clomifene group in the express of ovarian FSHRmRNA( P< 0.05) .
Conclusions: BSN decoction could overall regulation hypothalamus- pituitary-ovarian axis function of kidney-yang defieiency female rat model. BSN decoction could significant improve the weight of rat , level of the rat serum E2,FSH and LH;regulate the content of DA,NE and 5-HT;improve the histomorphology of rat ovarian, uterine and ultrastrucure of ovarian,uterine; improve the expression of hypothalamus Gondaotropin releasing hormone( GnRH);enhance the express of uterine,ovarian FSHR;enhance the express of ovarian FSHRmRNA.BSN decoction had the effect of inducing ovulation and recovering the function of ovary, the allover effect was better than clomifene.
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