钙调神经磷酸酶抑制剂对过氧化氢诱导的干细胞凋亡的影响  

Inhibitor of calcineurin protects stem cells from H_2O_2-induced apoptosis

作　　者：尹巧香[1] 王恒[2] 裴志勇[3] 赵玉生[4] 

机构地区：[1]中国人民解放军空军总医院干部病房心内科,北京100142 [2]中国人民解放军空军总医院神经内科,北京100142 [3]中国人民解放军北京军区总医院干部病房一科,北京100070 [4]中国人民解放军总医院老年心血管病研究所,北京100853

出　　处：《生物医学工程与临床》2014年第1期74-79,共6页Biomedical Engineering and Clinical Medicine

基　　金：中国科技部"863"课题(2006AA02A105)

摘　　要：目的比较钙调神经磷酸酶(CaN)抑制剂FK-506和环孢菌素A(CsA)保护脂肪组织来源干细胞(AT-DSCs)抗凋亡作用及可能机制。方法采用过氧化氢(H2O2)[分浓度0(对照组)、50、100、200、300、400μmol/L H2O26组]体外诱导AT-DSCs凋亡模型,凋亡模型的评价采用磷脂酰丝氨酸结合蛋白-异硫氰酸荧光素(AnnexinV-FITC)/碘化丙啶(PI)双染流式细胞术和线粒体内跨膜电位测定法;观察AT-DSCs凋亡细胞形态;AT-DSCs细胞活性检测采用CCK-8试剂;细胞色素C的表达采用Western blot分析。结果与对照组比较,引起细胞凋亡率最高的是浓度100μmol/L H2O2(15.60%±8.50%vs 4.50%±0.88%;P<0.01);但CaN抑制剂FK-506(质量浓度1mg/mL)(9.8%±1.3%vs17.0%±1.6%,P<0.01)和CsA(质量浓度1mg/mL)(12.2%±1.2%vs 17.0%±1.6%,P<0.01)预处理后细胞凋亡率均明显下降,细胞存活率明显提高(P<0.01)。Western blot结果显示,CaN抑制剂FK-506(质量浓度1 mg/mL)和CsA(质量浓度1 mg/mL)可完全抑制H2O2诱导的细胞色素C释放。结论 CaN抑制剂能保护H2O2诱导的干细胞凋亡。可能机制是CaN抑制剂能抑制细胞凋亡,提高细胞存活率,下调细胞色素C的释放。Objective To investigate the effect of inhibitor of calcineurin（CaN） in protecting the adipose tissue-derived stem cells （AT-DSCs） against apoptosis and the possible mechanisms. Methods A model of hydrogen peroxide（H202）-induced apoptosis was assessed by Annexin V-FITC/PI double staining cytometry and measurement of mitoehondria membrane potential. The morphology of apoptotic cells was observed. Cell activity was determined by CCK-8 assay. Expression of cytochrome C was investigated by Western blot method. Results Compared to the control, evidences showed that 100 txmol/L H202 could induce highest degree of apoptotic change in AT-DSCs（15.60 % ± 8.50 % vs 4.50 % ± 0.88 %, P〈 0.01）. This effect was inhibited by pre- treatment with calcineurin inhibitor FK-506 （1 mg/mL）（9.8 % ± 1.3 % vs 17.0 % ± 1.6 %, P〈 0.01） and CsA （1 mg/mL）（12.2 % ± 1.2 % vs 17.0 % ± 1.6 %, P 〈 0.01）, induced to significantly increased survival rate. Furthermore, Western blot analysis showed that FK-506（1 mg/mL） and CsA（1 mg/mL） completely inhibited the H^Og-induced release of cytochrome C. Conclusion These resuhs suggest that the inhibitor of calcineurin could protect the H202-induced stem cell apoptosis. The possible mechanisms may be related to the decreased apoptosis rate, increase cell activity and down -regulation of the release of cytochrome C.

关 键 词：钙调神经磷酸酶 干细胞 细胞凋亡 抑制剂 

分 类 号：R542.22[医药卫生—心血管疾病]