HuR诱导骨髓间充质干细胞成软骨分化的作用探析

HuR induces bone marrow mesenchymal stem cells into chondrogenic differentiation

目的观察HuR对骨髓间充质干细胞（bone marrow mesenchymal stem cell,BMSC）成软骨分化的影响,并初步探讨其作用机制。方法选取4周龄SD大鼠提取骨髓,制备BMSC,取第3代BMSC,分为HuR组和NC组,分别用pAd Easy-HuR-GFP病毒液（HuR组）和pAd Easy-GFP病毒液（NC组）干预,诱导14 d后进行固定、石蜡包埋、切片。行甲苯胺蓝进行染色,免疫组化、qPCR和检测XIAP、HuR、Col2a1的表达差异。结果甲苯胺蓝染色结果显示过表达HuR组的蛋白聚糖阳性信号比NC组多,且软骨球直径更大。免疫组化结果显示,XIAP、HuR、Col2a1随着含药血清浓度的提高逐渐上调,XIAP上调趋势比较明显,HuR和Col2a1上调不是很明显。qPCR检测到软骨HuR、XIAP明显升高,col2a1上调不明显。结论过表达HuR有利于BMSC分化成软骨,其机制可能与HuR过表达后调控XIAP转录增加有关。

Objective To observe the effect of HuR on chondrogenic differentiation of bone marrow mesenchymal stem cells（BMSCs） and to explore its mechanism. Methods BMSCs were extracted from 4-week-old SD rats. The third generation BMSCs were divided into a HuR group, and a NC group, and were respectively treated with pAd Easy-HuR-GFP virus solution and pAd Easy-GFP virus solution. After intervention, the cells were fixed 14 days after induction, paraffin-embedded and sectioned. Toluidine blue staining, QPCR and IHC detection of XIAP, HuR, Col2a1 expression differences were performed. Results Toluidine blue staining showed that positive signals of proteoglycan staining by HuR staining were more than NC and cartilage ball diameter was larger than NC. QPCR revealed that cartilage HuR, XIAP was significantly increased, Col2a1 up-regulation was not obvious. Immunohistochemical results showed that XIAP, HuR, Col2a1 gradually increased with the increase of drug-containing serum concentration, XIAP upward trend more obvious, HuR and Col2a1 upregulation is not obvious. QPCR detected cartilage HuR,XIAP was significantly increased, Col2a1 up-regulation is not obvious. Conclusion Overexpression of HuR facilitates the differentiation of BMSCs into cartilage, which may be related to the regulation of XIAP transcription after HuR over-expression.