發(fā)布時間：2018-01-27 08:52

  本文關鍵詞： 激素性股骨頭缺血性壞死 巨噬細胞移動抑制因子 血管修復 體外實驗　出處：《中國修復重建外科雜志》2016年08期 　論文類型：期刊論文

【摘要】：目的通過檢測體外缺氧/激素作用下內(nèi)皮細胞(endothelial cells,ECs)增殖、遷徙以及巨噬細胞移動抑制因子(macrophage migration inhibitory factor,MIF)和VEGF的表達變化,探討激素性股骨頭缺血性壞死(steroid-induced avascular necrosis of femoral head,SANFH)血管修復障礙發(fā)生機制。方法取第3代人臍靜脈ECs(human umbilical vein ECs,HUVECs)進行分組干預。正常對照組(A組)細胞不作任何干預;地塞米松組(B組):用含1.0×10~(-6) mol/L地塞米松的完全培養(yǎng)液干預;缺氧培養(yǎng)組(C組):細胞在低氧細胞培養(yǎng)罐內(nèi)培養(yǎng);地塞米松+缺氧培養(yǎng)組(D組):細胞在低氧細胞培養(yǎng)罐內(nèi)培養(yǎng),同時用含1.0×10~(-6) mol/L地塞米松的完全培養(yǎng)液干預。干預培養(yǎng)后24 h,行Alamar Blue細胞活性檢測及活/死細胞染色觀測細胞增殖情況,細胞骨架染色觀察細胞骨架形態(tài);采用劃痕實驗比較各組細胞遷徙能力,ELISA法檢測各組細胞MIF及VEGF表達水平。結(jié)果干預培養(yǎng)后24 h,Alamar Blue細胞活性檢測及活/死細胞染色示C組細胞活性最好且活細胞數(shù)多,D組細胞活性最差、活細胞數(shù)最少,組間比較差異均有統(tǒng)計學意義(P0.05)。細胞骨架染色見,A、B組細胞形態(tài)正常;C組細胞大量增殖,細胞內(nèi)見大量分泌顆粒;D組細胞骨架形態(tài)異常。劃痕實驗示C組細胞遷移能力最強,D組最弱。各組MIF和VEGF累積濃度隨時間延長均顯著增高。各時間點,C組MIF累積濃度顯著高于其他各組(P0.05)。干預培養(yǎng)24 h內(nèi),各組組內(nèi)1~8 h時MIF階段濃度顯著低于0~1 h及8~24 h時(P0.05);且C組0~1 h和8~24 h時MIF階段濃度顯著高于其他組(P0.05)。干預培養(yǎng)2 h內(nèi),各組組內(nèi)0.5~1 h MIF階段濃度顯著高于其他各時間段(P0.05)。8、24 h時C組VEGF累積濃度顯著高于其他各組(P0.05)。干預24 h內(nèi),C、D組8~24 h時VEGF階段濃度顯著高于其他各時間段(P0.05);干預培養(yǎng)2 h內(nèi),各組組內(nèi)各時間段及組間VEGF階段濃度比較,差異均無統(tǒng)計學意義(P0.05)。結(jié)論缺氧條件下ECs增殖及遷徙能力增強,MIF及VEGF分泌增加;而高濃度地塞米松會抑制上述過程,引發(fā)血管修復障礙,導致SANFH發(fā)生、發(fā)展。
[Abstract]:Objective to detect the proliferation of endothelial cells (ECs) induced by hypoxia / hormone in vitro. The expression of macrophage migration inhibitory factor MIF and VEGF in migration and macrophage migration suppressor. To investigate steroid-induced avascular necrosis of femoral head. Methods the third generation of human umbilical vein ECs(human umbilical vein ECs was used. HUVECs were divided into groups. The normal control group (group A) was not treated with any intervention. Dexamethasone group (group B) was treated with mol/L dexamethasone (1.0 脳 10 ~ (-1)) dexamethasone. Anoxic culture group (group C): cells were cultured in hypoxic cell culture tank; Dexamethasone anoxic culture group (group D): cells were cultured in hypoxic cell culture tank. At the same time, mol/L dexamethasone containing 1.0 脳 10 ~ (-1) -6) was used for 24 h after intervention. The activity of Alamar Blue cells was detected and the cell proliferation was observed by living / dead cell staining. The cytoskeleton morphology was observed by cytoskeleton staining. The expression of MIF and VEGF in the cells of each group were detected by scratch test and Elisa. Results 24 hours after intervention, the expression of MIF and VEGF were detected. Alamar Blue cell activity test and live / dead cell staining showed that C group had the best cell activity, and group D had the worst cell activity and the number of living cells was the least. The difference between the two groups was statistically significant (P 0.05). The cytoskeleton staining showed that the morphology of the cells was normal in group B. In group C, a large number of cells proliferated and a large number of secretory granules were found in the cells. The morphology of cytoskeleton in group D was abnormal. Scratch test showed that the ability of cell migration in group C was the strongest and the accumulation of MIF and VEGF in group D was the weakest. The cumulative concentrations of MIF and VEGF increased significantly with time. The cumulative concentration of MIF in group C was significantly higher than that in other groups (P 0.05). The concentration of MIF at 1h and 8h in each group was significantly lower than that at 0 h and 824 h, respectively. The concentration of MIF in group C was significantly higher than that in other groups at 0 h and 824 h. The concentration of 0. 5 h MIF in each group was significantly higher than that in other time periods (P 0. 05. 8). The cumulative concentration of VEGF in group C was significantly higher than that in other groups at 24 h. The concentration of VEGF phase in group D was significantly higher than that in other time periods at 824 h. Within 2 hours of intervention, there was no significant difference in the concentration of VEGF in each time period and between groups. Conclusion the proliferation and migration ability of ECs was enhanced under hypoxia condition. [WT5 "HZ] [WT5" HZ] [WT5 "BZ] [WT5" BZ]. The secretion of MIF and VEGF increased. High concentration of dexamethasone can inhibit these processes, causing vascular repair disorders, leading to the occurrence and development of SANFH.
【作者單位】： 西安交通大學第二附屬醫(yī)院骨一科;
【基金】：國家自然科學基金資助項目(81301562) 西安交通大學第二附屬醫(yī)院人才培養(yǎng)專項科研基金資助項目[RC(XM)201501]~~
【分類號】：R681.8
【正文快照】： 激素性股骨頭缺血性壞死(steroid-induced avas-cular necrosis of femoral head,SANFH)是骨科常見病、多發(fā)病[1]。近年研究發(fā)現(xiàn),SANFH臨床標本中可觀察到嚴重微血管損傷,骨細胞缺血性壞死,新生血管減少,血管修復障礙等病理現(xiàn)象[2-4]。糖皮質(zhì)激素(glucocorticoids,Glc)誘發(fā)的

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