骨髓间充质干细胞联合活性水凝胶治疗2型糖尿病大鼠心肌梗死研究

Co-transplantation of bone marrow derived-mesenchymal stem cells and bioactive hydrogel for myocardium infarction therapy in type 2 diabetic rats

目的利用水凝胶作为骨髓间充质干细胞(BM-MSCs)移植的载体,治疗2型糖尿病大鼠急性心肌梗死。方法 SD大鼠20只,3周龄,体质量50 g左右,用于制造2型糖尿病模型。将造模成功的SD大鼠随机分为水凝胶联合细胞移植组、细胞移植组、生理盐水组、假手术组,每组5只。将水凝胶联合细胞移植组、细胞移植组、生理盐水组2型糖尿病大鼠冠状动脉左前降支结扎,制造急性心肌梗死模型,同时做不结扎的假手术组。前3组分别移植100μL Matrigel和1×106 BM-MSCs的混合物、100μL含有1×106 BM-MSCs的0.9%氯化钠溶液、100μL 0.9%氯化钠溶液。术后28 d,检测大鼠血糖、血脂及血胰岛素水平;对术后28 d的大鼠心脏样本进行组织学染色评价心室重构情况;v WF(von Willebrand factor)免疫荧光染色观察大鼠心脏梗死区血管新生情况。结果 SD大鼠的BM-MSCs呈典型的长梭形且贴壁生长;经过8周高脂高糖饲料喂养后,血样生物化学指标显示,SD大鼠血液中总胆固醇、低密度脂蛋白、胰岛素等显著升高;后注射链脲霉素,大鼠空腹血糖在7.0 mmol/L以上,说明成功建立糖尿病大鼠模型;BM-MSCs移植糖尿病大鼠体内后,与不治疗组相比,BM-MSCs移植能显著改善心肌梗死后大鼠左心室重构;免疫荧光染色结果显示,BM-MSCs能促进梗死区内的血管新生。联合Matrigel移植能进一步提升BM-MSCs的治疗效果。结论联合Matrigel移植能显著提升BM-MSCs对糖尿病大鼠心肌梗死的治疗效果。

Objective To evaluate the therapeutic effect of co-transplantation of bone marrow derived-mesenchymal stem cells（BM-MSCs） with Matrigel for treatment of acute myocardial infarction（AMI） in type 2 diabetic（T2DM） rats. Methods T2 DM models were induced with 3-week-old SD rats（n = 20） with body weight of 50 g, which were randomly divided into 4 groups（n = 5）： hydrogel co-transplantation group, cell transplantation group, 0.9 % sodium chloride solution group and sham operation group. The left anterior descending coronary artery of T2 DM SD rats were permanently ligated, and the rats suffered AMI received transplantation of 100 μL mixture of Matrigel and 1 × 106BM-MSCs（hydrogel co-transplantation group）, 1 × 106BM-MSCs suspended in 100 μL 0.9 % sodium chloride solution（cell transplantation group）, and 100 μL 0.9 % sodium chloride solution（0.9 % sodium chloride solution group）, with the sham operation group as experimental control. The blood glucose, serum lipid and insulin of rats were measured at 28-day postoperation. All of rats were sacrificed and the heart remodeling was evaluated by histological staining. Immunofluorescent staining was performed to detect the blood vessels reconstruction in MI heart. Results The BM-MSCs isolated from SD rats were typically long fusiform and demonstrated adhesion dependent growth. The level of total cholesterol, low density lipoprotein cholesterol, and insulin in serum of SD rats were obviously up-regulated after 8-week high fat and high fructose diet. After Streptozotocin injection,the fasting blood-glucose of SD rats was all above 7.0 mmol/L, which indicated that the diabetic rat models were successfully constructed. Compared with the saline solution group, transplantation of BM-MSCs obviously improved the heart remodeling post MI,which was favorable for angiogenesis in ischemic heart as determined by immunofluorescent staining. Furthermore, co-transplantation with Matrigel was improved the therapeutic effect of BM-MSCs for MI. Conclusion It is demonstrated that co-transplantation with Matrigel could significantly improve the therapeutic effect of BM-MSCs for MI in T2 DM rats.