中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2013年
6期
963-968
,共6页
毛洪波%邹赛%唐俊明%杨建业%王家宁%孔霞%郭凌郧%郑飞%张蕾%黄永章
毛洪波%鄒賽%唐俊明%楊建業%王傢寧%孔霞%郭凌鄖%鄭飛%張蕾%黃永章
모홍파%추새%당준명%양건업%왕가저%공하%곽릉운%정비%장뢰%황영장
干细胞%骨髓干细胞%间充质干细胞%H9C2 细胞%心肌细胞%心肌梗死%基质细胞源衍生因子 1%凋亡%PI3-K/Akt%国家自然科学基金%干细胞图片文章
榦細胞%骨髓榦細胞%間充質榦細胞%H9C2 細胞%心肌細胞%心肌梗死%基質細胞源衍生因子 1%凋亡%PI3-K/Akt%國傢自然科學基金%榦細胞圖片文章
간세포%골수간세포%간충질간세포%H9C2 세포%심기세포%심기경사%기질세포원연생인자 1%조망%PI3-K/Akt%국가자연과학기금%간세포도편문장
背景:有研究显示表达 CXCR4的干细胞能够沿着基质细胞衍生因子1的浓度梯度迁移到心肌梗死部位再生心肌和血管而改善心脏的功能.目的:探索间充质干细胞通过其分泌的基质细胞衍生因子1对心肌细胞的保护作用.方法:收集培养2 d 的间充质干细胞条件培养基.在缺氧条件,利用基质细胞衍生因子1受体 CXCR4阻断剂 AMD3100或 PI3-K/Akt 途径阻断剂 LY294002预处理 H9C2细胞后,利用 AnnexinV/PI 双标法流式细胞术分析间充质干细胞条件培养基作用下 H9C2细胞凋亡的变化;Western blotting 分析 H9C2细胞磷酸化 Akt 蛋白的表达;RT-PCR 分析间充质干细胞基质细胞衍生因子1的表达.结果与结论:RT-PCR 结果显示间充质干细胞表达基质细胞衍生因子1,Western blotting 结果显示间充质干细胞条件培养基增加了 H9C2细胞磷酸化 Akt 蛋白的水平.AnnexinV/PI 分析发现间充质干细胞条件培养基明显降低了 H9C2细胞缺氧复氧后的凋亡,且这种抗凋亡作用能被 CXCR4阻断剂 AMD3100或 PI3-K/Akt 途径阻断剂 LY294002所阻断.说明间充质干细胞通过其分泌的基质细胞衍生因子1通过激活 PI3-K/Akt 途径保护 H9C2细胞,增加 H9C2细胞的幸存能力.
揹景:有研究顯示錶達 CXCR4的榦細胞能夠沿著基質細胞衍生因子1的濃度梯度遷移到心肌梗死部位再生心肌和血管而改善心髒的功能.目的:探索間充質榦細胞通過其分泌的基質細胞衍生因子1對心肌細胞的保護作用.方法:收集培養2 d 的間充質榦細胞條件培養基.在缺氧條件,利用基質細胞衍生因子1受體 CXCR4阻斷劑 AMD3100或 PI3-K/Akt 途徑阻斷劑 LY294002預處理 H9C2細胞後,利用 AnnexinV/PI 雙標法流式細胞術分析間充質榦細胞條件培養基作用下 H9C2細胞凋亡的變化;Western blotting 分析 H9C2細胞燐痠化 Akt 蛋白的錶達;RT-PCR 分析間充質榦細胞基質細胞衍生因子1的錶達.結果與結論:RT-PCR 結果顯示間充質榦細胞錶達基質細胞衍生因子1,Western blotting 結果顯示間充質榦細胞條件培養基增加瞭 H9C2細胞燐痠化 Akt 蛋白的水平.AnnexinV/PI 分析髮現間充質榦細胞條件培養基明顯降低瞭 H9C2細胞缺氧複氧後的凋亡,且這種抗凋亡作用能被 CXCR4阻斷劑 AMD3100或 PI3-K/Akt 途徑阻斷劑 LY294002所阻斷.說明間充質榦細胞通過其分泌的基質細胞衍生因子1通過激活 PI3-K/Akt 途徑保護 H9C2細胞,增加 H9C2細胞的倖存能力.
배경:유연구현시표체 CXCR4적간세포능구연착기질세포연생인자1적농도제도천이도심기경사부위재생심기화혈관이개선심장적공능.목적:탐색간충질간세포통과기분비적기질세포연생인자1대심기세포적보호작용.방법:수집배양2 d 적간충질간세포조건배양기.재결양조건,이용기질세포연생인자1수체 CXCR4조단제 AMD3100혹 PI3-K/Akt 도경조단제 LY294002예처리 H9C2세포후,이용 AnnexinV/PI 쌍표법류식세포술분석간충질간세포조건배양기작용하 H9C2세포조망적변화;Western blotting 분석 H9C2세포린산화 Akt 단백적표체;RT-PCR 분석간충질간세포기질세포연생인자1적표체.결과여결론:RT-PCR 결과현시간충질간세포표체기질세포연생인자1,Western blotting 결과현시간충질간세포조건배양기증가료 H9C2세포린산화 Akt 단백적수평.AnnexinV/PI 분석발현간충질간세포조건배양기명현강저료 H9C2세포결양복양후적조망,차저충항조망작용능피 CXCR4조단제 AMD3100혹 PI3-K/Akt 도경조단제 LY294002소조단.설명간충질간세포통과기분비적기질세포연생인자1통과격활 PI3-K/Akt 도경보호 H9C2세포,증가 H9C2세포적행존능력.
BACKGROUND: Several studies have demonstrated that stem cells expressing CXCR4 can migrate into myocardial infarction region to improve the heart function by regenerating myocardial tissue and vessels under the gradient concentrations of stromal cel -derived factor-1. OBJECTIVE: To investigate the protective effect of stromal cel -derived factor-1 from bone marrow mesenchymal stem cells on myocardial cells. METHODS: Conditioned medium of bone marrow mesenchymal stem cells cultured for 2 days was col ected. Under hypoxic condition, H9C2 cells were pretreated with CXCR4 inhibitor AMD3100 (5 mg/mL) or PI3-K/Akt inhibitor LY294002 (10 μM) for 1 hour. After treated with bone marrow mesenchymal stem cel conditioned medium, H9C2 cel apoptosis was analyzed by Annexin V/PI double staining methods. Expression of Akt and pAkt in H9C2 cells was analyzed by Western blotting. Expression of stromal cel -derived factor-1 was analyzed by RT-PCR. RESULTS AND CONCLUSION: RT-PCR results showed that bone marrow mesenchymal stem cells expressed stromal cel -derived factor-1. Western blotting results showed that bone marrow mesenchymal stem cel conditioned medium increased pAkt protein level in H9C2 cells. Annexin V/PI analysis showed that bone marrow mesenchymal stem cel conditioned medium significantly decreased apoptosis of H9C2 cells induced by hypoxia/reoxygenation and this anti-apoptotic effect could be blocked by CXCR4 inhibitor AMD3100 or PI3-K/Akt inhibitor LY294002. Stromal cel -derived factor-1 from mesenchymal stem cells plays an important role in the protection of cardiomyocytes through PI3-K/Akt signal pathway.
