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pAdEasy-HuR-GFP病毒液(HuR组)和pAdEasy-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 pAdEasy-HuR-GFP virus solution and pAdEasy-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.
  / RNA结合蛋白 / HuR / BMSC / 成软骨分化
RNA binding protein; HuR;   / BMSC / Differentiation into cartilage
[1] Zhuang Z, Ye G, Huang B. Kaempferol alleviates the interleukin-1β-induced inflammation in rat osteoarthritis chondrocytesvia suppression of NF-κB[J]. Med Sci Monit, 2017, 14(23): 3925-3931.
[2] Jia S, Zhang T, Xiong Z, et al. In vivo evaluation of a novel oriented scaffold-BMSC construct for enhancing full-thickness articular cartilage repair in a rabbit model[J]. PLoS One, 2015, 10(12) :e0145667
[3] Mostafavinia A, Dehdehi L, Ghoreishi SK, et al. Effect of in vivo low-level laser therapy on bone marrow-derived mesenchymal stem cells in ovariectomy-induced osteoporosis of rats[J]. J Photochem Photobiol B, 2017,18(175):29-36.
[4] Cao XK, Li R, Sun W, et al. Co-combination of islets with bone marrow mesenchymal stem cells promotes angiogenesis[J]. Biomed Pharmacother, 2016, 78:156-164. [5] Chang N, Ge J, Xiu L, Zhao Z, et al. HuR mediates motility of human bone marrow-derived mesenchymal stem cells triggered by sphingosine 1-phosphate in liver fibrosis[J]. J Mol Med (Berl), 2017, 95(1): 69-82.
[6] Alcaraz MJ, Megías J, García-Arnandis I, et al. New molecular targets for the treatment of osteoarthritis. Biochem Pharmacol, 2010, 80(1):13-21.
[7] Han SH, Kim YH, Park MS, et al. Histological and biomechanical properties of regenerated articular cartilage using chondrogenicbone marrow stromal cells with a PLGA scaffold in vivo[J]. J Biomed Mater Res A, 2008,87(4):850-861.
[8] Chen FH, Tuan RS. Mesenchymal stem cells in arthritic diseases[J]. Arthritis Res Ther, 2008,10(5):223.
[9] Grammatikakis I, Abdelmohsen K, Gorospe M. Posttranslational control of HuR function[J]. Wiley Interdiscip Rev RNA, 2017, 8(1): 1372-1385.
[10]Ge J, Chang N, Zhao Z, et al. Essential roles of RNA-binding protein HuR in activation of hepatic stellate cells induced by transforming growth factor-β1[J]. Sci Rep, 2016, 25(6): 22141-22155.
[11] Zhang X, Zou T, Rao JN, et al. Stabilization of XIAP mRNA through the RNA binding protein HuR regulated by cellular polyamines[J]. Nucleic Acids Res, 2009 , 37(22): 7623-7637.
[12]Abdel-Magid AF. Modulation of the inhibitors of apoptosis proteins (IAPs) activities for Cancer Treatment[J]. ACS Med Chem Lett, 2017, 8(5): 471-473.
[13]Hou Y, Allan L A, Clarke PR. Phosphorylation of XIAP by CDK1-cyclin-B1 controls mitotic cell death[J]. J Cell Sci, 2017, 130(2): 502-511.
[14] Prabhu K S, Siveen K S, Kuttikrishnan S, et al. Targeting of X-linked inhibitor of apoptosis protein and PI3-kinase/AKT signaling by embelin suppresses growth of leukemic cells[J]. PLoS One, 2017, 12(7): e180895.
[15] Hikami S, Shiozaki A, Kitagawa-Juge M, et al. The role of cIAP1 and XIAP in apoptosis induced by tumor necrosis factor alpha in esophageal squamous cell carcinoma cells[J]. Dig Dis Sci, 2017, 62(3): 652-659.
[16] Bohm B, Hess S, Krause K, et al. ADAM15 exerts an antiapoptotic effect on osteoarthritic chondrocytes via up-regulation of the X-linked inhibitor of apoptosis[J]. Arthritis Rheum, 2010, 62(5): 1372-1382.
[17] Simons M, Beinroth S, Gleichmann M, et al. Adenovirus-mediated gene transfer of inhibitors of apoptosis protein delays apoptosis in cerebellar granule neurons[J]. J Neurochem, 1999, 72(1): 292-301.
国家自然科学基金(81403421)
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