补肾填精法对化疗源性卵巢早衰大鼠生育力保存的影响

Effects of Nourishing Kidney to Replenish Marrow Therapy on Fertility of Rats with Chemotherapy-induced Premature Ovarian Failure

【目的】探讨补肾填精法代表方左归丸对环磷酰胺(CTX)致化疗源性卵巢早衰(POF)大鼠生育力保存的影响。【方法】将121只SPF级成熟雌性SD大鼠随机分为空白组(N=8),模型组(N=18),POF左归丸低、中、高剂量组(均N=18),POF达菲林组(N=17),非POF左归丸组(N=8),非POF达菲林组(N=16);另取雄性SD大鼠20只交配用。造模前的第10天当天,POF达菲林组、非POF达菲林组大鼠给予肌肉注射达菲林药液,其他组别肌肉注射生理盐水;造模期间,模型组,POF左归丸低、中、高剂量组,POF达菲林组腹腔注射CTX建立化疗源性POF大鼠模型,其他组别腹腔注射生理盐水,连续15 d;造模后,POF左归丸低、中、高剂量组及非POF左归丸组分别给予相应药物灌胃,空白组、模型组、POF达菲林组、非POF达菲林组给予等体积过滤水灌胃,连续灌胃30、60 d。后雌雄合笼交配,通过检测阴栓及精子筛选受孕大鼠,并解剖受孕大鼠,观察子宫情况,统计胚胎着床情况,计算受孕率。【结果】(1)灌胃30、 60 d后受孕率:与空白组比较,模型组灌胃30、 60 d后受孕率均明显下降(P<0.05);与模型组比较,POF左归丸中、高剂量组,非POF达菲林组,非POF左归丸组灌胃30 d后受孕率,非POF达菲林组、非POF左归丸组灌胃60 d后受孕率明显上升(P<0.05)。(2)累计受孕率:与空白组比较,模型组,POF左归丸低、高剂量组,POF达菲林组累计受孕率降低(P<0.05);与模型组比较,POF左归丸中、高剂量组,非POF达菲林组,非POF左归丸组累计受孕率均高于模型组(P<0.05)。(3)裸眼观察大鼠子宫形态:空白组子宫宫体红润,胚胎着床位点分布相对均匀;模型组大鼠子宫颜色苍白,宫体细长,胚胎着床位点量少且分布不均匀;而POF左归丸各剂量组大鼠子宫宫体红润,胚胎着床位点分布较空白组稀疏且不均匀,但较模型组数量明显增多。【结论】CTX化疗会导致大鼠生育能力降低,而左归丸可改善CTX化疗源性POF大鼠的生育能力。

Objective To explore the effect of Zuogui Pills(representative recipe for nourishing kidney to replenish marrow therapy)on the fertility of rats with chemotherapy drug cyclophosphamide(CTX)-induced premature ovarian failure(POF).Methods One hundred and twenty-one SPF mature female SD rats were randomly divided into blank group(N=8),model group(N=18),POF low-,middle-and high-dose Zuogui Pills groups(N=18,respectively),POF Diphereline group(N=17),non-POF Zuogui Pills group(N=8),and non-POF Diphereline group(N=16).Additionally,there were 20 male SD rats used for mating.On day 10 before modeling,POF Diphereline group and non-POF Diphereline group were given intramuscular injection of Diphereline liquid,and the other groups were given intramuscular injection of normal saline.And then during the modeling period,the rats in the model group,POF low-,middle-and high-dose Zuogui Pills groups and POF Diphereline group were given intraperitoneal injection of CTX to induce into chemotherapy-induced POF model,and the others were given intraperitoneal injection of normal saline.After modeling,POF low-,middle-and high-dose Zuogui Pills groups and non-POF Zuogui Pills group were respectively given intragastric administration of corresponding drug,and the blank group,model group,POF Diphereline group and non-POF Diphereline group were given intragastric administration of the same volume of normal saline.After treatment for 30,60 days,half of the female rats in various groups were allocated to one cage for mating with male rats.And then we screened out the pregnant rats by vaginal plug and sperms determination,and dissected to observe the morphological characteristics of the uterus.The embryo implantation was ascertained,and the pregnancy rate was calculated.Results(1)The pregnancy rate after treatment for 30,60 days:compared with the blank group,the pregnancy rate in the model group after treatment for 30,60 days was decreased(P<0.05).Compared with the model group,the pregnancy rate in the POF middle-and high-dose Zuogui Pills groups,non-POF Diphereline group,nonPOF Zuogui Pills group after treatment for 30 days and in non-POF Diphereline group,non-POF Zuogui Pills group after treatment for 60 days was increased(P<0.05).(2)The accumulative total pregnancy rate:compared with the blank group,the accumulative total pregnancy rate in the model group,POF low-and high-dose Zuogui Pills groups,and POF Diphereline group was decreased(P<0.05);compared with the model group,the accumulative total pregnancy rate in the POF middle-and high-dose Zuogui Pills groups,non-POF Diphereline group,and non-POF Zuogui Pills group was increased(P<0.05).(3)The morphological characteristics of uterus observed by naked eyes:the uterus body was ruddy,the distribution of embryo implantation was relatively uniform in the blank group;the uterus body was pale,long and thin,the number of embryo implantation site was less and was distributed unevenly in the model group;the uterus body was ruddy,the embryo implantation sites were sparse and uneven in various POF Zuogui Pills groups as compared with those in the blank group,but was more than the model group.Conclusion CTX can induce the fortility of rats to decrease,but Zuogui Pills have the effects on improving the fertility of chemotherapy-induced POF in rats.