摘要
目的:探讨尼古丁作用人脐带间充质干细胞(MSCs)前后,一氧化氮(NO)、细胞内钙离子(Ca2+)的变化及α7nAchR的表达情况。方法:采用硝酸还原酶法检测不同质量浓度尼古丁作用于MSCs后24、36、48 h,MSCs内NO的释放情况;尼古丁作用MSCs 24 hf,luo-3/AM染色,流式细胞仪检测MSCs内Ca2+变化;用RT-qPCR检测尼古丁作用后α7nAchR的表达情况。结果:尼古丁作用MSCs 24、36 h后,各实验组NO水平显著高于对照组(P<0.05),呈时间、质量浓度依赖性,但在48 h,质量浓度0.8与1.0 mg/mL组,NO水平低于对照组。尼古丁作用后,各实验组质量浓度(0.6、0.8、1.0 mg/mL)的Ca2+荧光强度分别为(141.26±16.01)(、164.90±18.39)、(198.76±17.63),均高于对照组(119.30±14.14),其中质量浓度0.8、1.0 mg/mL组与对照组相比有显著差异(P<0.05)。MSCs可表达α7nAchR,且尼古丁作用后表达水平增高,呈时间浓度依赖性。结论:尼古丁可上调人脐带MSCs的α7nAchR表达,并通过刺激MSCs释放NO、升高细胞内Ca2+从而致MSCs凋亡。
Aim：To explore the effects of nicotine on intracellular nitric oxide, Calcium ion and ex-pression of ot7nAchR in umbilical cord mesenchymal stem cells. Methods： MSCs were treated with dif-ferent concentrations of nicotine. The level of NO was detected by nitrate reductase method at 24,36,48h ;after cultured with nicotine for 2gh, cells were stained using by fluo-3/AM and the change of Ca^2＋ wasdetermined by flow cytometer. The expression of α7 nAchR was detected by Real-time Quantitative PCR.Results： After treated with nicotine, the nitric oxide level in each experimental group were significantlyhigher than that of the control group at 24h and 36h（ P 〈0. 05 ）, in a dose and time dependent manners,however at 48 h the levels of NO in 0. 8 mg/mL group andl. 0 mg/mL group were lower than the controlgroup, which may be related with the increased cells apoptosis. After treated with nicotine, the Ca^2＋ fluo-rescence intensity in each experimental group （ 0. 6, 0. 8, 1.0 mg/mL） were （ 141.26 ±. 16. 01 ）,（ 164. 90±18.39）, （ 198.76±17.63）, respectively,which were higher than the control group （ 119.30±14. 14）. The expression of otTnAchR was detected in MSCs, and was found to be up-regulated aftertreated with nicotine in a dose-dependent manner and a time-dependent manner. Conclusion：Nicotinecan promote MSCs apoptosis by up-regulating the expression of ct7nAchR, stimulating MSCs to releaseNO and elevating the intracellular Ca^2＋.
出处
《暨南大学学报（自然科学与医学版）》
CAS
CSCD
北大核心
2011年第6期598-601,共4页
Journal of Jinan University(Natural Science & Medicine Edition)
基金
国家自然科学基金项目(30973127)
广州市科技攻关项目(11BPPZXaa2070004)
