丹参酮ⅡA对骨髓间充质干细胞心肌方向分化间隙连接蛋白43表达的干预作用

Effects of Tanshinone ⅡA on Connexin 43 Expression During Mesenchymal Stem Cells Differentiating into Cardiac Muscle Cells

【目的】研究丹参酮ⅡA对骨髓间充质干细胞(MSCs)心肌方向分化间隙连接蛋白43(Cx43)表达的干预作用。【方法】体外培养大鼠MSCs,采用逆转录聚合酶链反应(RT-PCR)、Western blotting和免疫荧光技术检测不同组间MSCsCx43表达。【结果】未分化的MSCs上Cx43有弱的表达;5-氮胞甙(5-aza)、正常心肌上清液以及缺氧心肌上清液诱导后Cx43的表达显著升高(P<0.01);丹参酮高剂量组(1.5×10-7 mol/L)Cx43的表达量显著增加(P<0.05),丹参酮低剂量组(1.5×10-8mol/L)也显著升高(P<0.01);经缺氧心肌上清液和丹参酮共同干预后,Cx43表达较对照组均显著升高,但丹参酮低剂量组作用更加显著(P<0.01);缺氧心肌上清液和丹参酮低剂量组联合干预之后Cx43表达接近正常心肌细胞组(P>0.05),各组间荧光强度的变化与RT-PCR和Western blotting的结果一致。【结论】不同浓度丹参酮ⅡA可上调MSCs上Cx43表达,但丹参酮ⅡA和缺氧心肌上清液联合诱导作用更强。

Objective To observe the effect of tanshinone ⅡA （T ⅡA） on the connexin （ Cx43 ） expression during mesenchymal stem cells （MSCs） differentiating into cardiac muscle cells in vitro. Methods MSCs were cultured in vitro, and Cx43 protein and mRNA expression levels were detected with reverse transcriptase polymerase chain reaction （RT-PCR） , Western blotting and immunofluorescence staining methods. Results The results of RT-PCR showed that Cx43 expression level in undifferentiated MSCs was low, and then was increased after induced by 5- aza, normal myocardial supernatant, and hypoxia myocardial supernatant, respectively （P 〈 0. 01 ）. The effect on Cx43 expression varied in different concentrations of T ⅡA, Cx43 expression level was increased after treated by 1.5 ×10^7 mol/L T ⅡA （P 〈 0. 05） and 1.5 ×10^8 mol/L T ⅡA （P 〈 0. 01 ）. The Cx43 expression was increased to normal level after treated together by 1.5×10^8 mol/L T ⅡA and hypoxia myocardial supernatant （ P 〉 0.05 ）. The change of fluorescent intensity in various groups was the same as that shown by RT-PCR and Western blotting method. Conclusion Various concentrations of T ⅡA can upregulate the Cx43 expression during MSCs differentiating into cardiac muscle cells, and the induction effect is the best after treatment with the combination of T ⅡA and hypoxia myocardial supernatant.