HMGB1联合MSCs移植对急性心肌梗死大鼠心脏血管生成影响

Effects of HMGB1 in combination with MSCs transplantation on angiogenesis after acute myocardial infarction in rats

目的通过高迁移率组蛋白1(High Mobility Group Box-1,HMGB1)预处理骨髓间充质干细胞(Mesenchymal stem cells,MSCs),观察HMGB1能否增强MSCs旁分泌效应;在大鼠急性心肌梗死的模型上,HMGB1同时联合移植MSCs,观察HMGB1联合MSCs移植是否能进一步增加梗死区域局部新生血管数量。方法贴壁离心法培养大鼠MSCs,HMGB1的受体TLR4和RAGE检测;不同浓度梯度HMGB1分别干预MSCs后,ELISA法检测VEGF的表达情况;SD雄性大鼠分为4组:模型组、MSCs移植组、HMGB1注射组、HMGB1注射+MSCs移植组(n=24只),制备大鼠急性心肌梗死模型;术后3、7、28dELISA法检测血清VEGF浓度水平;术后28d免疫组化测定梗死区新生血管密度。结果 1MSCs有TLR4和RAGE的表达;2浓度为12.5、25、50、100和200ng/mL HMGB1干预MSCs后,VEGF的分泌较对照组显著增加,当浓度为400和800ng/mL时,VEGF的分泌量较对照组减少(P﹤0.05);3术后3、7d时间点血清VEGF测定:HMGB1注射+MSCs移植组>MSCs移植组>HMGB1注射组>模型组(P<0.05);4HMGB1注射+MSCs移植组的CD31染色新生血管数明显增多。结论 HMGB1联合MSCs移植对急性心肌梗死大鼠心梗区及交界区新生血管生成有益。

Objective This study is to examine whether mesenchymal stem cells （MSCs） can producebetter effects by injecting High Mobility Group Box - 1 （ HMGB1 ） in rat model of acute myocardial infarction, focusingonthe new- born vesselsin the heart infarct area. Methods To obtain high purity rat MSCs by side -wall centrifugation, and the TLR4 and RAGE of HMGB1 detection. SD male rats were randomly divided into four groups：model control group, MSCs transplantation group, HMGB1 injection group, HMGB1 injection ＋ MSCs trans - plantation group （ n = 24）. Acute myocardial infarction for rats was established hy open heart surgey. The VEGF levels were examined 3, 7 and 28 days of operation by ELISA. The density of newborn blood vessels in the infarct area after the operation for 28 days was examined. Results 1 ） Isolated MSCs expressed TLR4 and RAGE receptor. 2）HMGB1 at concentrations of 12.5, 25, 50, 100 and 200 ng/mL added to MSCs cultures increased the secretion of VEGF, while at higher concentrations of 400 and 800 ng/mL, VEGF secretion was - decreased compared to control group. 3） 3 and 7 days after acute myocardial infarction, serum VEGF levels were in the order of HMGB1 injection ＋ MSCs transplantation group 〉 MSCs transplantation group 〉 HMGB1 injection group 〉 model control group. 4） In HMGB1 injection ＋ MSCs transplantation group, CD31 staining of new blood vessels was increased obviously. Conclusion HMGB1 combing MSCs transplantation is useful for the treatment for acute myocardial infarction in rats by increasing local new blood vessels.