摘要
目的研究缩泉丸含药血清对小鼠胚胎干细胞(ESCs)增殖、分化及Nanog基因表达的影响。方法选用129小鼠胚胎干细胞株,将培养的细胞分为空白组,金匮肾气丸组,缩泉丸低、中、高剂量组5组。空白组给予浓度为10%的空白大鼠血清;金匮肾气丸组给予浓度为10%的含药血清;缩泉丸低、中、高剂量组分别给予按0.585 g·kg^(-1)·d^(-1)、1.17 g·kg^(-1)·d^(-1)、2.34 g·kg^(-1)·d^(-1)剂量灌胃给药的大鼠的含药血清,浓度均为10%。采用流式细胞仪检测缩泉丸对ESCs增殖的影响,倒置相差显微镜观察缩泉丸含药血清对DMSO诱导的ESCs的分化情况,荧光定量PCR检测ESCs关键因子Nanog mRNA的表达,流式法检测Nanog蛋白的表达。结果与空白组比较,缩泉丸含药血清中、高剂量能够促进ESCs的增殖(P<0.01),缩泉丸低、高剂量可以显著提高ESCs中Nanog mRNA的表达(P<0.05,P<0.01),缩泉丸中、高剂量可以显著提高ESCs中Nanog蛋白的表达(P<0.05,P<0.01)。与空白组比较,缩泉丸高剂量能够促进DMSO诱导的ESCs进行分化。结论缩泉丸可以促进ESCs的增殖,促进DMSO诱导的ESCs进行分化,可能与其上调关键基因Nanog的表达相关。
Objective To study the effects of drug-containing serum of Suoquan Pill on proliferation and differentiation of mouse embryonic stem cells( ESCs) and Nanog gene expression. Methods The embryonic stem cell line derived from 129 mice was selected. The cultured cells were divided into the blank group,Jingui Shenqi Pill group,Suoquan Pill groups with low-,middle-and high-dose. The blank group was treated with blank serum of rats at concentration of 10%,the Jingui Shenqi Pill group was treated with the drug-containing serum at concentration of 10%. The drug-containing serum of Suoquan Pill was obtained from rats that treated with Suoquan Pill for intragastric administration at daily dose of 0. 585,1. 17 and 2. 34 g·kg^(-1)·d^(-1)respectively. The Suoquan Pill groups with low-,middle-and high-dose were treated with the corresponding drug-containing serum at concentration of 10%. The effect of Suoquan Pill on ESCs proliferation was detected by flow cytometry. The effect of drug-containing serum of Suoquan Pill on differentiation of DMSO induced ESCs was observed by inverted phase contrast microscope. The Nanog mRNA expression in ESCs was detected by fluorescent quantitative PCR and the Nanog protein expression was detected by flow cytometry.Results Compared with the blank group,the drug-containing serum of Suoquan Pill with middle-and high-dose could promote ESCs proliferation( P < 0. 01),the drug-containing serum of Suoquan Pill with low-and high-dose could significantly increase Nanog mRNA expression in ESCs( P < 0. 05,P < 0. 01),and the drugcontaining serum of Suoquan Pill with middle-and high-dose could significantly up-regulate Nanog protein expression in ESCs( P < 0. 05,P < 0. 01). Compared with the blank group,the drug-containing serum of Suoquan Pill with high-dose could promote the differentiation of DMSO induced ESCs. Conclusion Suoquan Pill can promote ESCs proliferation and differentiation of DMSO induced ESCs,which may be associated with the up-regulation of Nanog expression.
出处
《上海中医药杂志》
2017年第8期93-97,共5页
Shanghai Journal of Traditional Chinese Medicine
基金
教育部博士点科研基金资助项目(20104425120002)
福建省中青年教师教育科研项目(JA14423)
