摘要
目的探讨组织激肽释放酶1(KLK1)诱导大鼠骨髓间充质干细胞(BMSCs)向心肌样细胞分化的影响。方法选取2015年3—7月购买的大鼠BMSCs,将大鼠BMSC细胞分为3组即正常组、组织激肽释放酶1组、5-Aza组。采用RT-PCR分别对诱导培养后的大鼠BMSCs进行心肌细胞特异性蛋白Actin、c-Tn I和Desmin的检测,并用Western blot免疫印迹分析c-Tn T蛋白表达量。结果 RT-PCR检测显示,5-Aza诱导的BMSCs(1.964±0.805)在心肌特异性蛋白Actin RNA、c-Tn I RNA和Desmin RNA的表达均高于空白组(1.000±0.000)(P<0.05),KLK1诱导组(2.633±0.582)以上蛋白的RNA表达量又显著高于5-Aza诱导组(P<0.05)。Western blot结果显示,5-Aza诱导组c-Tn T蛋白的表达(0.799±0.101)高于空白组(0.112±0.001)(P<0.001),KLK1诱导组(1.294±0.134)显著高于5-Aza组。差异有统计学意义(P<0.001)。结论 5-Aza能有效使BMSCs向心肌样细胞诱导分化,KLK1也能诱导BMSCs向心肌样细胞分化。KLK1较5-Aza诱导的BMSCs在心肌细胞特异性蛋白RNA表达,心肌特异性蛋白T表达其心肌特异性更为突出。
[Abstract] Objective To study on effect of ex p res sion vector with KLK1 in d u c in g th e BMSCs c a rd ia c - l ik e ce l l d if fe re n t ia -tion. Methods Select th e ra t BMSC ce lls from March to July 2 01 5 were d iv id ed into th e normal group, KLK1 group an d 5- Aza group, and the rat BMSCs cells were tes ted by the cardiac cel l specif ic protein Act in, c-TnI and Desmin by the RT- PCR, and the c-TnT protein expres sion level was analyzed by the Western blot immunoblotting. Results T he R T -P C R te s t showed that the expres sion of BMSCs induced by 5-Aza in the myocardial marker protein Act in RNA, c-TnI RNA andDesmin RNA was higher than that in the blank group[ （1.964±0.805） vs （1.000±0.000）]（P〈0.05）, and the RNA expression level of proteins above the （2.633±0.582） in the KLK1 induct ion group was obviously higher than that in the 5-Aza induc-tion group （P 〈 0 .0 5 ） , an d th e W e s te rn blot re su lts showed th a t th e c -T n T pro tein ex p res sion leve l in th e 5 -A z a in -d uc tio n group was h ig he r th an th a t in th e b lan k group,[ （0 .7 9 9± 0 .1 0 1 ） vs （0 .1 1 2 ± 0 .0 0 1 ） ] （P 〈 0 .0 0 1 ） , an d th e d if-fe ren c e s had s ta t is t ic a l sign if ic an ce . Conclusion 5 -Aza c a n effectively m ake th e BMSCs c a rd ia c - l ik e ce l l differ-entiat ion, KLKlalso can induce the BMSCs cardiac-like cel l different iation, and the cardiac- speci f ici ty of KLK1 in the BM- SCs induced by the KLK1 cardiac cel l specif ic protein RNA expres sion and myocardial marker protein T expres sion is more prominent than induced by the 5-Aza.
出处
《中外医疗》
2017年第3期11-15,共5页
China & Foreign Medical Treatment
基金
国家自然科学基金项目(NO.81260597)
甘肃中医药大学甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室
关键词
心肌细胞
骨髓间充质干细胞
组织激肽释放酶1
肌动蛋白
结蛋白
肌钙蛋白
Cardiac mu sc le cell
Bone marrow derived mesen ch ymal stem cell
T is su e k a l l ik re in l
Actin
Desmin
Tro-ponin
