奥美拉唑对小鼠精原干细胞生长与增殖分化的影响

The effect of omeprazole enteric-coated tablets on growth and proliferation of spermatogonial stem cells in mouse

目的探讨抑酸药物(奥美拉唑)对小鼠精原干细胞(SSCs)生长与增值分化的影响。方法以5～7日龄昆明小白鼠为材料,体外分离、培养小鼠睾丸支持细胞及精原干细胞,以支持细胞为饲养层,培养小鼠SSCs。研究分实验组和空白对照组,在共培养8 h后,实验组添加奥美拉唑(质子泵抑制剂) 1μmol/L,对照组不添加任何物质。在培养24、72、120、192、240 h分别用显微镜直视下观察两组SSCs在不同时间点SSCs生长形态及数目;在培养24、72、120、192、240 h后采用MTT法分别检测两组小鼠SScs生长与增值情况;培养240 h后运用PCR技术检测两组SSCS DNA含量。结果小鼠SSCs培养6～8 h开始贴壁生长,两组细胞呈透亮、圆形,体积中等,共培养240 h时,对照组细胞形成数百个细胞团,实验组细胞则数目相对较少。培养24、72、120、192、240 h时,实验组小鼠SSCs吸光度(OD)值均低于对照组;实验组内不同时间点OD值比较,差异有统计学意义(P <0. 05); PCR结果:对照组基因DNA量明显高于实验组(P <0. 05)。结论抑酸药物对小鼠SSCs生长与增值分化有明显的抑制作用,抑酸药物可能会影响男性的生殖能力。

Objective To investigate the effects of omeprazole enteric-coated tablets( omeprazole) on the growth and proliferation of mice spermatogonial stem cells( SSCs).Methods SSCs and supporting cells were isolated from 5 ~ 7-day-old Kunming mice.The supporting cells were used as feeder layers to culture SSCs.The SSCs were divided into experimental group and blank control group.After 8 h of co-culture,the experimental group was given omeprazole( proton pump inhibitor) 1 μmol/L,while the control group was not added with omeprazole.The growth morphology and number of SSCs were observed under microscope at 24 h,72 h,120 h,192 h and 240 h.After 24 h,72 h,120 h,192 h and 240 h,the growth and proliferation of SSCs in the two groups were detected by using MTT assay.After 240 hours of culture,the SSCs DNA content of the two groups was detected by PCR.Results SSCs began to adhere to the wall at 6-8 h of the culture. The cells in the two groups were bright,round and of medium size. The cells in the control group formed tens to hundreds of cell clusters after 240 hours of culture,and the number of cells in the experimental group was small.When co-cultured for24 h,72 h,120 h,192 h and 240 h,the absorbance values of SSCs in the experimental group were lower than those in the control group( P < 0. 05).PCR results showed that the amount of DNA in the experimental group was significantly lower than that in the control group( P < 0. 05).Conclusion Acid-suppressing drugs can significantly inhibit the growth and proliferation of mice SSCs.Therefore,acid-suppressing drugs may affect male reproductive ability.