中国草食动物科学2025，Vol.45Issue(6)：41-48,8.DOI:10.3969/j.issn.2095-3887.2025.00.028
催乳素对绒山羊附睾上皮细胞增殖和凋亡的影响
Effect of Prolactin on Proliferation and Apoptosis of Epididymal Epithelial Cells in Cashmere Goats
摘要
Abstract
The epididymis is the site of sperm maturation and storage.Epididymal epithelial cells secrete epididymal fluid to promote the development and maturation of sperm.The proliferation and differentiation of epididymal epithe-lial cells are regulated by reproductive hormones,however whether prolactin(PRL)is involved in this process remains to be studied.This experiment studied the effects of PRL on the proliferation,migration and apoptosis of cashmere goat epididymal epithelial cells through in vitro culture.After the epididymal tissue samples of Yanshan cashmere goats were collected sterilely,the epididymal epithelial cells were isolated by enzymatic digestion,followed by purified by differential adherence and secondary trypsin digestion,and identified by immunofluorescence.The purified epididymal epithelial cells were randomly divided into 7 groups with 6 replicates in each group,and 0(control group),25,50,75,100,200,and 400 ng/mL PRL were administrated to the culture medium,respectively.The cell proliferation activity was detected by CCK-8 method at 12 h,24 h,and 48 h during culture.Followed by the optimal concentration and time of PRL to promote the proliferation of epididymal epithelial cells were determined,the cell scratch experiment,flow cy-tometry and real-time flurescence quantitative PCR(qRT-PCR)were employed to detect cell migration,apoptosis,as well as apoptotic genes expression level.The results showed that the proliferative activity of epididymal epithelial cells was significantly promoted in the 25,50,75,and 100 ng/mL PRL groups treated for 12 h compared with the control group(P<0.05),and the highest proliferative activity was observed in the 50 ng/mL PRL group,while the proliferative activity of cells in the 200 and 400 ng/mL PRL groups did not differ significantly from that of the control group(P>0.05).After cultured for 24 h and 48 h,the proliferation activity of cells in 25,50,and 75 ng/mL PRL groups was not significantly different from that in the control group(P>0.05),while the proliferation activity of cells in 100,200,and 400 ng/mL PRL groups was significantly lower than that in the control group(P<0.05).The results of cell scratch ex-periment showed that the cell migration distance and cell migration rate of the 50 ng/mL PRL group treated for 12 h were significantly higher than those of the control group(P<0.05),the total apoptosis rate and the expression levels of apoptosis genes Bax and Caspase-3 were significantly lower than those in the control group(P<0.05),and the expres-sion level of anti-apoptosis gene Bcl-2 was significantly higher than that in the control group(P<0.05).In conclusion,the addition of appropriate concentrations of PRL(50 ng/mL)and treatment of cells for 12 h significantly promoted the proliferation and migration of epididymal epithelial cells and inhibited cell apoptosis.关键词
绒山羊/催乳素/附睾上皮细胞/增殖/凋亡Key words
cashmere goats/prolactin/epididymal epithelial cells/proliferation/apoptosis分类
畜牧业引用本文复制引用
刘晓娜,段春辉,郑庆丰,王会杰,左艳华,李祥龙,刘月琴,张英杰..催乳素对绒山羊附睾上皮细胞增殖和凋亡的影响[J].中国草食动物科学,2025,45(6):41-48,8.基金项目
农业生物育种重大专项(2023ZD0405102) （2023ZD0405102）
国家绒毛用羊产业技术体系(CARS-39) （CARS-39）
中央引导地方科技发展资金项目(246Z6602G) （246Z6602G）
