中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
28期
4530-4534
,共5页
滕金龙%李丹%于海初%蔡尚郎%潘新亭
滕金龍%李丹%于海初%蔡尚郎%潘新亭
등금룡%리단%우해초%채상랑%반신정
干细胞%移植%脓毒症%骨髓干细胞%骨髓间充质干细胞%重组人促红细胞生成素%心肌损伤%大鼠%造血干细胞移植%凋亡
榦細胞%移植%膿毒癥%骨髓榦細胞%骨髓間充質榦細胞%重組人促紅細胞生成素%心肌損傷%大鼠%造血榦細胞移植%凋亡
간세포%이식%농독증%골수간세포%골수간충질간세포%중조인촉홍세포생성소%심기손상%대서%조혈간세포이식%조망
erythropoietin%mesenchymal stem cells%sepsis%myocytes,cardiac%inflammation
背景：相关研究表明促红细胞生成素及骨髓间充质干细胞均对心肌凋亡有一定影响，但两者联合应用治疗脓毒症相关性心肌损伤少见报道。目的：观察骨髓间充质干细胞移植联合腹腔内注射促红细胞生成素对脓毒症大鼠心肌细胞病理学改变及细胞凋亡的影响。方法：选用SD大鼠50只，随机数字表法均分为5组(n=10)，应用盲肠结扎穿孔术建立脓毒症大鼠模型。骨髓间充质干细胞组建模后即刻尾静脉输注异体间充质干细胞；促红细胞生成素组建模后即刻腹腔内注射促红细胞生成素；促红细胞生成素+细胞移植组两者联合应用；模型组行盲肠结扎穿孔术，对照组开腹后不做任何处理，均尾静脉输注相同容量的生理盐水。24 h 后麻醉处死实验动物，取心肌标本，采用苏木精-伊红染色观察心肌组织形态；用Western blot方法检测心肌组织中凋亡蛋白Bax、Caspase-3，抗凋亡蛋白Bcl-2的表达量。结果与结论：苏木精-伊红染色结果：对照组可见心肌细胞排列整齐，心肌细胞结构完整；模型组可见广泛心肌纤维断裂，排列紊乱，心肌细胞肿胀或皱缩，可见空泡变性；心肌间质血管充血、水肿，炎症细胞浸润；促红细胞生成素组与骨髓间充质干细胞组心肌组织相似，炎性细胞浸润情况较轻，期间散在分布正常心肌细胞；促红细胞生成素+细胞移植组心肌细胞损害较轻，间质充血不明显，少量炎症细胞浸润。Western blot结果：促红细胞生成素+细胞移植组Bcl-2蛋白表达显著高于促红细胞生成素组、模型组及对照组(P <0.01)， Bax及Caspase-3蛋白表达量均减低(P<0.05)。结果显示，骨髓间充质干细胞移植联合给予促红细胞生成素在脓毒症相关性心肌损伤的治疗中可减轻心肌的病理学改变、抑制心肌细胞的凋亡，其机制可能是通过上调抗凋亡蛋白、下调凋亡蛋白的表达来实现的。
揹景：相關研究錶明促紅細胞生成素及骨髓間充質榦細胞均對心肌凋亡有一定影響，但兩者聯閤應用治療膿毒癥相關性心肌損傷少見報道。目的：觀察骨髓間充質榦細胞移植聯閤腹腔內註射促紅細胞生成素對膿毒癥大鼠心肌細胞病理學改變及細胞凋亡的影響。方法：選用SD大鼠50隻，隨機數字錶法均分為5組(n=10)，應用盲腸結扎穿孔術建立膿毒癥大鼠模型。骨髓間充質榦細胞組建模後即刻尾靜脈輸註異體間充質榦細胞；促紅細胞生成素組建模後即刻腹腔內註射促紅細胞生成素；促紅細胞生成素+細胞移植組兩者聯閤應用；模型組行盲腸結扎穿孔術，對照組開腹後不做任何處理，均尾靜脈輸註相同容量的生理鹽水。24 h 後痳醉處死實驗動物，取心肌標本，採用囌木精-伊紅染色觀察心肌組織形態；用Western blot方法檢測心肌組織中凋亡蛋白Bax、Caspase-3，抗凋亡蛋白Bcl-2的錶達量。結果與結論：囌木精-伊紅染色結果：對照組可見心肌細胞排列整齊，心肌細胞結構完整；模型組可見廣汎心肌纖維斷裂，排列紊亂，心肌細胞腫脹或皺縮，可見空泡變性；心肌間質血管充血、水腫，炎癥細胞浸潤；促紅細胞生成素組與骨髓間充質榦細胞組心肌組織相似，炎性細胞浸潤情況較輕，期間散在分佈正常心肌細胞；促紅細胞生成素+細胞移植組心肌細胞損害較輕，間質充血不明顯，少量炎癥細胞浸潤。Western blot結果：促紅細胞生成素+細胞移植組Bcl-2蛋白錶達顯著高于促紅細胞生成素組、模型組及對照組(P <0.01)， Bax及Caspase-3蛋白錶達量均減低(P<0.05)。結果顯示，骨髓間充質榦細胞移植聯閤給予促紅細胞生成素在膿毒癥相關性心肌損傷的治療中可減輕心肌的病理學改變、抑製心肌細胞的凋亡，其機製可能是通過上調抗凋亡蛋白、下調凋亡蛋白的錶達來實現的。
배경：상관연구표명촉홍세포생성소급골수간충질간세포균대심기조망유일정영향，단량자연합응용치료농독증상관성심기손상소견보도。목적：관찰골수간충질간세포이식연합복강내주사촉홍세포생성소대농독증대서심기세포병이학개변급세포조망적영향。방법：선용SD대서50지，수궤수자표법균분위5조(n=10)，응용맹장결찰천공술건립농독증대서모형。골수간충질간세포조건모후즉각미정맥수주이체간충질간세포；촉홍세포생성소조건모후즉각복강내주사촉홍세포생성소；촉홍세포생성소+세포이식조량자연합응용；모형조행맹장결찰천공술，대조조개복후불주임하처리，균미정맥수주상동용량적생리염수。24 h 후마취처사실험동물，취심기표본，채용소목정-이홍염색관찰심기조직형태；용Western blot방법검측심기조직중조망단백Bax、Caspase-3，항조망단백Bcl-2적표체량。결과여결론：소목정-이홍염색결과：대조조가견심기세포배렬정제，심기세포결구완정；모형조가견엄범심기섬유단렬，배렬문란，심기세포종창혹추축，가견공포변성；심기간질혈관충혈、수종，염증세포침윤；촉홍세포생성소조여골수간충질간세포조심기조직상사，염성세포침윤정황교경，기간산재분포정상심기세포；촉홍세포생성소+세포이식조심기세포손해교경，간질충혈불명현，소량염증세포침윤。Western blot결과：촉홍세포생성소+세포이식조Bcl-2단백표체현저고우촉홍세포생성소조、모형조급대조조(P <0.01)， Bax급Caspase-3단백표체량균감저(P<0.05)。결과현시，골수간충질간세포이식연합급여촉홍세포생성소재농독증상관성심기손상적치료중가감경심기적병이학개변、억제심기세포적조망，기궤제가능시통과상조항조망단백、하조조망단백적표체래실현적。
BACKGROUND:Erythropoietin and bone marrow mesenchymal stem cells have been shown to affect myocardial apoptosis. However, few studies concerned their combined application to sepsis-related myocardial injury. OBJECTIVE:To observe the effects of the combination of erythropoietin and bone marrow mesenchymal stem cells on pathology and apoptosis of sepsis rat cardiomyocytes. METHODS:A total of 50 Sprague-Dawley rats were randomly selected and assigned to five groups (n=10). Sepsis models were established by cecal ligation perforation method. Rat models in the bone marrow mesenchymal stem cellgroup, erythropoietin group and erythropoietin+bone marrow mesenchymal stem cellgroup were respectively treated with bone marrow mesenchymal stem cells, erythropoietin and erythropoietin+bone marrow mesenchymal stem cells immediately after model induction via intraperitoneal injection or caudal vein. Model group received cecum ligation and puncture. Control group did not undergo any treatment after the abdomen was opened. Model and control groups were infused with an equal volume of physiological saline via caudal vein. At 24 hours, experimental animals were sacrificed by anesthesia. Myocardial specimens were col ected. Myocardial appearance was observed using hematoxylin-eosin staining. Bax, Caspase-3 and Bcl-2 were tested by western blot assay. RESULTS AND CONCLUSION:Hematoxylin-eosin staining:cardiomyocytes were regularly arranged, showing integrated structure in the control group. Extensive myocardial fiber breakage, disordered arrangement, cardiomyocyte swel ing or shrinkage, and vacuolar degeneration were observed in the model group. Moreover, myocardial interstitial vascular congestion, edema, and inflammatory cellinfiltration were visible. Myocardial tissue was similar between erythropoietin and bone marrow mesenchymal stem cellgroups, with the presence of mild inflammatory cellinfiltration and scattered normal cardiomyocytes. In the erythropoietin+bone marrow mesenchymal stem cellgroup, cardiomyocytes were slightly damaged. Interstitial vascular congestion was not apparent, and a few inflammatory cells infiltrated. Western blot assay results demonstrated that Bcl-2 protein expression was significantly higher (P<0.01), but Bax and Caspase-3 protein expression was lower (P<0.05) in the erythropoietin+bone marrow mesenchymal stem cellgroup than in the erythropoietin, model and control groups. The combination of erythropoietin and bone marrow mesenchymal stem cells in treatment of sepsis-related myocardial injury could lessen myocardial pathological changes, and inhibit cardiomyocyte apoptosis. The mechanisms maybe exert by upregulating anti-apoptotic protein expression and downregulating apoptotic protein expression.
