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远细胞(TCs)是一种新型间质细胞,分布于机体的多个组织和器官,参与组织的再生和修复.但是,TCs在血管中的功能及作用机制尚未见相关报道.本研究通过建立大鼠(Rattus norvegicus)颈总动脉球囊损伤(CABI)模型,利用透射电子显微镜(TEM)及免疫荧光双染CD34/vimentin检测模型中TCs的变化;于体外分离培养血管TCs,探索TCs的作用及机制.免疫荧光定量分析显示,与假手术组(Sham组)相比,损伤组(CABI组)TCs数目明显增多((7.2±1.0)/mm~2 vs(20.4±1.8)/mm~2,P<0.001).取TCs培养上清孵育血管平滑肌细胞(vascular smooth muscle cells,VSMCs),经流式细胞仪分析VSMCs的细胞周期,发现与对照组(TCs-)相比,TCs培养上清孵育VSMCs(TCs+)48 h后,处于S期和G2/M期的VSMCs百分比升高约5%((10.68+3.53)%vs(13.51+5.46)%,(TCs-)vs(TCs+),P<0.05).原位杂交结果显示,血管TCs内miR-24的表达水平明显高于VSMCs,高出约3倍(P<0.001).预拮抗TCs内的miR-24后,TCs培养上清对VSMCs的促增殖效应减弱,表现为与对照组相比,处于S期和G2/M期VSMCs细胞百分比明显减少,分别为(12.76+4.92)%vs(9.89+2.85)%,(antagomir-NC vs antagomir-24,P<0.05).上述结果表明,血管TCs能够促进VSMCs的增殖,并且miR-24参与了TCs介导的促VSMCs增殖作用.
Far cells (TCs) are a new type of interstitial cells distributed in many tissues and organs of the body and involved in the regeneration and repair of tissues.However, the function and mechanism of TCs in blood vessels have not been reported yet.In this study, (CABI) model of rat common carotid artery (Rattus norvegicus), the changes of TCs in the model were detected by transmission electron microscopy (TEM) and double immunofluorescence staining of CD34 / vimentin; TCs were isolated and cultured in vitro to explore the role of TCs The results of immunofluorescence quantitative analysis showed that the number of TCs in injured group (CABI group) was significantly higher than that in Sham group (7.2 ± 1.0) / mm ~ 2 vs (20.4 ± 1.8) / mm ~ 2 , P <0.001) .Cells cultured in supernatants of TCs were incubated with VSMCs, and the cell cycle of VSMCs was analyzed by flow cytometry. Compared with the control group (TCs-), TCs culture supernatants The percentages of VSMCs in S phase and G2 / M phase increased about 5% ((10.68 + 3.53)% vs (13.51 + 5.46)%, (TCs-) vs (TCs +) after 48 h incubation of VSMCs <0.05) .In situ hybridization results showed that the expression level of miR-24 in vascular TCs was significantly higher than that in VSMCs by about 3 times (P <0.001). After pretreatment with miR-24 in TCs, TCs culture Compared with the control group, the percentage of VSMCs in VSMCs in S phase and G2 / M phase were significantly decreased, which were (12.76 + 4.92)% vs (9.89 + 2.85)%, respectively. The antagomir- NC vs antagomir-24, P <0.05). These results suggest that vascular TCs can promote the proliferation of VSMCs, and miR-24 is involved in the TCs-mediated proliferation of VSMCs.