重庆医学
重慶醫學
중경의학
Chongqing Medicine
2015年
27期
3752-3755
,共4页
烧伤%间质干细胞%微管%迁移%PI3K/Akt%信号通路
燒傷%間質榦細胞%微管%遷移%PI3K/Akt%信號通路
소상%간질간세포%미관%천이%PI3K/Akt%신호통로
burns%mesenchymal stem cells%microtubules%migration%PI3K/Akt%signaling pathway
目的：探讨严重烧伤大鼠血清对骨髓间充质干细胞(BMSCs)迁移的影响及作用机制。方法建立严重烧伤大鼠模型,并制备严重烧伤大鼠血清。实验分为正常培养组(含10%胎牛血清,C 组)、烧伤血清组(含10%烧伤大鼠血清,B 组)、烧伤血清+阻断剂组(含10%烧伤大鼠血清+终浓度为10μmol/L 的 PI3K 信号通路抑制剂 LY294002,B+LY 组),MTT 法检测各组细胞活性；Transwell 小室检测各组细胞迁移情况；Western blot 检测各组细胞 p-Akt、Akt 蛋白表达；免疫荧光检测各组细胞微管结构。结果与 C 组比较,B 组烧伤血清处理24 h 后,BMSCs 细胞活性增高(P <0.01),p-Akt 水平升高(P <0.05),细胞微管结构出现解聚,迁移数量增高(P <0.001)；加入抑制剂后,与 B 组比较,B+LY 组 BMSCs 细胞活性降低(P <0.01),迁移数量降低(P <0.001),p-Akt 水平降低(P <0.05),细胞微管结构变化不明显。结论严重烧伤大鼠血清可促进 BMSCs 细胞迁移,可能与烧伤血清中细胞因子激活 PI3K/Akt 信号通路相关,进而引起 BMSCs 细胞微管结构变化,促进 BMSCs 迁移。
目的：探討嚴重燒傷大鼠血清對骨髓間充質榦細胞(BMSCs)遷移的影響及作用機製。方法建立嚴重燒傷大鼠模型,併製備嚴重燒傷大鼠血清。實驗分為正常培養組(含10%胎牛血清,C 組)、燒傷血清組(含10%燒傷大鼠血清,B 組)、燒傷血清+阻斷劑組(含10%燒傷大鼠血清+終濃度為10μmol/L 的 PI3K 信號通路抑製劑 LY294002,B+LY 組),MTT 法檢測各組細胞活性；Transwell 小室檢測各組細胞遷移情況；Western blot 檢測各組細胞 p-Akt、Akt 蛋白錶達；免疫熒光檢測各組細胞微管結構。結果與 C 組比較,B 組燒傷血清處理24 h 後,BMSCs 細胞活性增高(P <0.01),p-Akt 水平升高(P <0.05),細胞微管結構齣現解聚,遷移數量增高(P <0.001)；加入抑製劑後,與 B 組比較,B+LY 組 BMSCs 細胞活性降低(P <0.01),遷移數量降低(P <0.001),p-Akt 水平降低(P <0.05),細胞微管結構變化不明顯。結論嚴重燒傷大鼠血清可促進 BMSCs 細胞遷移,可能與燒傷血清中細胞因子激活 PI3K/Akt 信號通路相關,進而引起 BMSCs 細胞微管結構變化,促進 BMSCs 遷移。
목적：탐토엄중소상대서혈청대골수간충질간세포(BMSCs)천이적영향급작용궤제。방법건립엄중소상대서모형,병제비엄중소상대서혈청。실험분위정상배양조(함10%태우혈청,C 조)、소상혈청조(함10%소상대서혈청,B 조)、소상혈청+조단제조(함10%소상대서혈청+종농도위10μmol/L 적 PI3K 신호통로억제제 LY294002,B+LY 조),MTT 법검측각조세포활성；Transwell 소실검측각조세포천이정황；Western blot 검측각조세포 p-Akt、Akt 단백표체；면역형광검측각조세포미관결구。결과여 C 조비교,B 조소상혈청처리24 h 후,BMSCs 세포활성증고(P <0.01),p-Akt 수평승고(P <0.05),세포미관결구출현해취,천이수량증고(P <0.001)；가입억제제후,여 B 조비교,B+LY 조 BMSCs 세포활성강저(P <0.01),천이수량강저(P <0.001),p-Akt 수평강저(P <0.05),세포미관결구변화불명현。결론엄중소상대서혈청가촉진 BMSCs 세포천이,가능여소상혈청중세포인자격활 PI3K/Akt 신호통로상관,진이인기 BMSCs 세포미관결구변화,촉진 BMSCs 천이。
Objective To study the effects of severely burned rats serum on migration of BMSCs and mechanism.Methods Severely burned rats model was established,and the preparation of severely burned rats serum.Experimental groups:normal train-ing group(containing 10% fetal bovine serum,group C),burn serum group(containing 10% burns in the rat serum,group B),burn serum+blockers(10% burns in the rat serum+final concentration of 10 μmol/L PI3K signaling pathway inhibitor LY294002 train-ing,group B+LY).Activity of cells was examined with MTT;migration of cells was examined with Transwell chambers testing;protein expression of p-AKT/AKT was determined with Western blot;microtubule structure of cells was examined with immuno-fluorescence.Results Compared with group C,group B burn serum treatment after 24 h,BMSCs activity(P <0.01),p-AKT levels (P <0.05),increased migration quantity(P <0.001);cell microtubule structures appear rupture,after adding inhibitor,compared with group B,group B+LY BMSCs activity(P <0.01),to reduce the number of migration(P <0.001),p-lower AKT(P <0.05), cell microtubule structure similar to the normal group.Conclusion Severely burned rats serum can promote BMSCs migration,may burn serum cytokine activation of PI3K/AKT signal pathway,resulting in cell microtubule structure change,promote the migration of BMSCs.
