基于转录组学探讨林蛙油对卵泡发育障碍大鼠的影响

Transcriptomics-based Exploration of the Effects of Oviductus Ranae on Rats with Impaired Follicular Development

目的:通过转录组学角度分析林蛙油对环磷酰胺所致卵泡发育障碍的影响,为林蛙油的雌激素样作用机制研究提供充足的理论及实验依据。方法:筛选后的雌性Wistar大鼠60只,随机分为对照组(C)、模型组(M)、阳性药组(P)、林蛙油低(ORL)、高剂量组(ORH);除对照组外,其余大鼠均腹腔注射环磷酰胺建立卵泡发育障碍大鼠模型;除模型组外,其余大鼠灌胃相应药物,连续灌胃14~16 d后测定大鼠血清性激素含量;摘取卵巢和子宫称量记录,计算器官指数;一侧卵巢与子宫做HE染色计算各级卵泡数量及子宫内膜厚度,另一侧卵巢进行转录组学分析,筛选差异基因,对差异基因进行GO和KEGG富集分析。结果:大鼠卵泡发育障碍模型成功建立。与模型组大鼠相比,林蛙油低、高剂量组大鼠子宫湿重和子宫指数显著升高(P<0.05),孕酮和雌二醇含量显著升高(P<0.05);林蛙油低、高剂量组大鼠窦前卵泡与黄体数量显著升高(P<0.05),子宫内膜厚度显著增加(P<0.01),差异有统计学意义,林蛙油低剂量组大鼠窦状卵泡数量显著升高(P<0.05);转录组结果显示,林蛙油低、高剂量组与模型组之间差异基因分别有738、572个;GO富集分析分别富集到1790、1616个条目;KEGG富集分析主要集中在PI3K/Akt信号通路与B细胞受体信号通路。结论:林蛙油能有效改善环磷酰胺所致卵泡发育障碍大鼠的血清性激素水平、卵巢卵泡发育情况、子宫内膜萎缩等情况,其分子机制可能与PI3K/Akt、B细胞受体等通路及基因有关。

Objective:The effects of Oviductus Ranae(OR)on cyclophosphamide-induced follicular developmental disorders were analyzed from the perspective of transcriptomics,which provided sufficient theoretical and experimental basis for the study of estrogen-like mechanism of OR.Methods:Sixty screened female Wistar rats were randomly divided into control group(C),model group(M),positive group(P),OR low-dose(ORL)and high-dose group(ORH).Except for the control group,all rats were injected with cyclophosphamide intraperitoneally to establish the rat model of follicular development dysfunction.Except for the model group,the other rats were given corresponding drugs by gavage,and the serum sex hormone contents were determined after 14~16 days of continuous gavage.The ovaries and uterus were extracted and weighed for recording and calculating the organ index.One ovary and uterus were stained with HE staining to calculate the number of follicles at all levels and the thickness of the endometrium,while the other ovary was analyzed by transcriptome,and the differentially expressed genes were screened and analyzed by GO and KEGG enrichment analyses.Results:The results showed that the rat follicular development dysfunction model was successfully established.Compared with the rats in the M,the wet weight and index of the uterus of rats in the ORL and ORH significantly increased(P<0.05),and the contents of P and E2 significantly increased(P<0.05).The number of preantral follicles and corpus luteum of rats in the ORL and ORH increased significantly(P<0.05),and the thickness of endometrium increased significantly(P<0.01),with statistically significant differences,and the number of antral follicles in the ORL increased significantly(P<0.05).The transcriptome results showed that there were 738 and 572 differential genes between the ORL,ORH and M,respectively.GO enrichment analysis enriched 1790 and 1616 entries and the KEGG enrichment analysis was mainly focused on the PI3K/Akt signaling pathway and the B-cell receptor signaling pathway.Conclusion:OR can effectively improve serum sex hormone levels,ovarian follicular development,and endometrial atrophy in rats with cyclophosphamide-induced follicular dysfunction,and its molecular mechanism may be related to pathways and genes such as PI3K/Akt and B-cell receptor.