中华临床医师杂志（电子版）
中華臨床醫師雜誌（電子版）
중화림상의사잡지（전자판）
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2013年
18期
8270-8274
,共5页
王艾丽%曾彬%程新耀%严斐斐
王艾麗%曾彬%程新耀%嚴斐斐
왕애려%증빈%정신요%엄비비
间质干细胞%心肌梗死%血红素加氧酶-1%细胞因子类%血管新生
間質榦細胞%心肌梗死%血紅素加氧酶-1%細胞因子類%血管新生
간질간세포%심기경사%혈홍소가양매-1%세포인자류%혈관신생
Mesenchymal stem cells%Myocardial infarction%Heme oxygenase-1%Cytokines%Angiogenesis
目的：探讨血红素氧合酶-1（HO-1）基因修饰的骨髓间充质干细胞（MSCs）培养上清液对大鼠心肌梗死的治疗作用。方法 HO-1腺病毒或对照GFP腺病毒转染MSCs，Western blot检测HO-1的表达；流式细胞仪检测无氧无血清条件下HO-1-MSCs、GFP-MSCs、MSCs的凋亡；ELISA和RT-PCR检测无氧无血清条件下 HO-1-MSCs 细胞因子的分泌。收集 HO-1-MSCs、GFP-MSCs 在无氧无血清条件下的培养上清液，于结扎冠状动脉1 h后多点注射到心肌梗死区边缘，注射Control Medium为对照组。注射4 d后检测心功能变化，4周后取梗死区边缘心肌行Masson染色和免疫组化CD34染色。结果 HO-1转染的 MSCs可以稳定高效地表达HO-1蛋白（P＝0.01），HO-1-MSCs在无氧无血清条件下的凋亡率显著低于GFP-MSCs和MSCs（P＝0.01），且其在无氧无血清条件下HGF、bFGF、TGF-β、VEGF分泌水平显著增高（P＝0.02）；注射4 d后发现HO-1-MSCs上清液治疗组心功能各项指标均显著改善（P＜0.01）；4周后HO-1-MSCs上清液治疗组胶原面积明显减少（P＜0.01），且梗死区微血管数量也增加。结论 HO-1-MSCs分泌多种细胞因子，通过促进血管新生，减少胶原沉积以及减少心肌坏死，从而改善心功能以及急性心肌梗死后的左心室重构。
目的：探討血紅素氧閤酶-1（HO-1）基因脩飾的骨髓間充質榦細胞（MSCs）培養上清液對大鼠心肌梗死的治療作用。方法 HO-1腺病毒或對照GFP腺病毒轉染MSCs，Western blot檢測HO-1的錶達；流式細胞儀檢測無氧無血清條件下HO-1-MSCs、GFP-MSCs、MSCs的凋亡；ELISA和RT-PCR檢測無氧無血清條件下 HO-1-MSCs 細胞因子的分泌。收集 HO-1-MSCs、GFP-MSCs 在無氧無血清條件下的培養上清液，于結扎冠狀動脈1 h後多點註射到心肌梗死區邊緣，註射Control Medium為對照組。註射4 d後檢測心功能變化，4週後取梗死區邊緣心肌行Masson染色和免疫組化CD34染色。結果 HO-1轉染的 MSCs可以穩定高效地錶達HO-1蛋白（P＝0.01），HO-1-MSCs在無氧無血清條件下的凋亡率顯著低于GFP-MSCs和MSCs（P＝0.01），且其在無氧無血清條件下HGF、bFGF、TGF-β、VEGF分泌水平顯著增高（P＝0.02）；註射4 d後髮現HO-1-MSCs上清液治療組心功能各項指標均顯著改善（P＜0.01）；4週後HO-1-MSCs上清液治療組膠原麵積明顯減少（P＜0.01），且梗死區微血管數量也增加。結論 HO-1-MSCs分泌多種細胞因子，通過促進血管新生，減少膠原沉積以及減少心肌壞死，從而改善心功能以及急性心肌梗死後的左心室重構。
목적：탐토혈홍소양합매-1（HO-1）기인수식적골수간충질간세포（MSCs）배양상청액대대서심기경사적치료작용。방법 HO-1선병독혹대조GFP선병독전염MSCs，Western blot검측HO-1적표체；류식세포의검측무양무혈청조건하HO-1-MSCs、GFP-MSCs、MSCs적조망；ELISA화RT-PCR검측무양무혈청조건하 HO-1-MSCs 세포인자적분비。수집 HO-1-MSCs、GFP-MSCs 재무양무혈청조건하적배양상청액，우결찰관상동맥1 h후다점주사도심기경사구변연，주사Control Medium위대조조。주사4 d후검측심공능변화，4주후취경사구변연심기행Masson염색화면역조화CD34염색。결과 HO-1전염적 MSCs가이은정고효지표체HO-1단백（P＝0.01），HO-1-MSCs재무양무혈청조건하적조망솔현저저우GFP-MSCs화MSCs（P＝0.01），차기재무양무혈청조건하HGF、bFGF、TGF-β、VEGF분비수평현저증고（P＝0.02）；주사4 d후발현HO-1-MSCs상청액치료조심공능각항지표균현저개선（P＜0.01）；4주후HO-1-MSCs상청액치료조효원면적명현감소（P＜0.01），차경사구미혈관수량야증가。결론 HO-1-MSCs분비다충세포인자，통과촉진혈관신생，감소효원침적이급감소심기배사，종이개선심공능이급급성심기경사후적좌심실중구。
Objective To investigate the effect of the culture supernatants from MSCs overexpressing HO-1 gene on a rat myocardial infarction. Methods MSCs were cultured in vitro and transfected by Adv-HO-1 or Adv-GFP. Western blot were used to determine the expression of HO-1. FACS was used to determined the apoptosis of MSCs under the condition of hypoxia plus serum-free. ELISA and RT-PCR were peformed to assay the expression and secrection of HGF, bFGF, TGF-β and VEGF. The culture supernatants from HO-1-MSCs or GFP-MSCs were injected into the infarcted border zone, separately. Four days after injection, cardiac function was measured by ultrasound, and 4 weeks after injection, the myocardial infarct size was measured with Masson’s trichrome, and the CD34 in myocardial infarction area was detected by immunohistochemical method. Results HO-1-MSCs demonstrated HO-1 expression increase and were more resistant to hypoxia plus serum-free stimulation. Various growth factors, including HGF, bFGF, TGF-β and VEGF were produced by HO-1-MSCs, some of which were enhanced significantly under hypoxia plus serum-free stimulation. Four days after injection, the culture supernatants from HO-1-MSCs significantly improved the early cardiac function, and 4 weeks after injection, the culture supernatants from HO-1-MSCs significantly increased vascular regeneration, decreased the myocardium apoptosis, and decreased the collagen deposition. Conclusion HO-1-MSCs may improve cardiac function and left ventricular remodeling after acute myocardial infarction, which can secrete a variety of cytokines, promote angiogenesis, decrease collagen deposition and myocardial necrosis.
