摘要
目的:研究缺血条件下薯蓣皂甙预处理对骨髓间充质干细胞(MSCs)凋亡的影响,探讨中药穿山龙作为MSCs移植治疗缺血性心肌病辅助治疗的可行性。方法:采用无血清方式诱导MSCs凋亡。实验分5组:对照、无血清、低、中、高剂量干预组。分别采用终浓度为4mg·mL-1,40mg·mL-1,400mg·mL-1薯蓣皂甙预处理第4代MSCs,24h后更换无血清DMEM饥饿48h,TUNEL标记细胞凋亡;细胞免疫组化染色检测Bcl-2蛋白水平表达;RT-PCR检测凋亡抑制基因Bcl-2mRNA表达。结果:TUNEL标记阳性细胞率按对照组、低、中、高剂量组、无血清组依次升高,除后两组,其余各组间差异有统计学意义;无血清干预48h后,Bcl-2蛋白表达增加,低、中剂量薯蓣皂甙预处理可进一步提高Bcl-2表达,但高剂量干预组与无血清组间无统计学差异(P=0.631);对照组Bcl-2mRNA表达显著低于各组(均为P<0.001),而高剂量干预组与无血清组间无差异(P=0.67);低剂量干预组Bcl-2基因片段mRNA表达最高,中剂量干预组次之,提示无血清条件下可提高Bcl-2基因表达,低、中剂量的薯蓣皂甙预处理可提高无血清诱导的MSCs的Bcl-2基因表达,但高剂量薯蓣皂甙预处理则无此作用。结论:适当剂量薯蓣皂甙预处理能使缺血条件下MSCs凋亡减少,使抗凋亡基因Bcl-2基因及蛋白表达增加。
Objective：To evaluate the effects of Dioscin （DIO）, a traditional Chinese medicine on apoptosis of mesenchymal stem cell （MSCs）. The feasibility of MSCs transplantation treating cardiomyopathy with DIO as an ancillary medicine. Methods： MSCs were cultured with serum deprivation and with Dioscin pretreatement in a different dose. The cultured MSCs were at randora divided into five groups： normal control group, serum deprivation group, serum deprivation with different dose of DIO intervention group （low dose group： 4 mg/L, moderate dose group： 40 mg/L, high dose group 400 mg/L）. The cultured cells were deprived serum for 48 hours. Myocardial apoptosis was assayed with the method of terminal deoxynulceotidyl transferase （TdT）-mediated dUTP-biotin nick end labeling （TUNEL）. Results：Compared with control group, the PEIs of TUNEL cells were significantly higher than that in serum group and each dose DIO groups （P〈0. 001 ）. The highest PEI was presented at serum deprivation group, and then was high dose group （P = 0. 024）. There was no significant difference between the above groups. After pretreatment with DIO, the PEI were significantly decreased compared with that in serum deprivation group. The PEI was significantly higher as the dose got higher （P〈0. 001 ）. There was a basement expression of Bcl-2 in control group. Compared with control group, The PEI of expressing Bcl-2 cells were significantly higher in the rest groups （P〈 0.001 ）. DIO pretreatment could increase the expressions of Bcl-2. The optional effect presented at low dose DIO,and then was moderate dose DIO. There was no significantly difference between the high dose group and serum deprivation group （P = 0. 061 ）. The effect promoting Bcl-2 expression by using intervention with low dose and moderate dose DIOwas more significant than that of serum deprivation. The expressions of Bcl-2 mRNA increased significantly in the serum deprivation group and different DIO dose group except high dose group （P
出处
《中国临床医学》
北大核心
2008年第6期904-905,共2页
Chinese Journal of Clinical Medicine
基金
大连市科委科研基金项目
关键词
薯蓣皂甙
凋亡
中药
骨髓间充质干细胞
Dioscin
Apoptosis
Chinese traditional medicine
Mesenchymal stem cell
