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目的观察缺氧预处理骨髓间充质干细胞(MSCs)移植对脑缺血再灌注损伤大鼠SDF-1/CXCR4 m RNA和蛋白表达的影响及清热化瘀方的干预作用。方法采用线栓法制备局灶性脑缺血大鼠(MCAO)模型,将216只SD大鼠随机分为6组:假手术组(SO)、模型组(MCAO)、MSCs移植对照组(N-MSCs)、经HP处理后的MSCs移植组(HP-MSCs)、MSCs移植联合清热化瘀方组(MSCs+QRHY)、HP-MSCs移植联合清热化瘀方组(HP+QRHY)。每组大鼠36只,每组根据取材时间点7,14,28 d又可分为每组3个亚组,每个亚组12只大鼠。采用q RT-PCR和Western blot观察3个时间点SDF-1/CXCR4 m RNA及其蛋白的表达变化,并以TUNEL法检测神经细胞凋亡。结果各组缺血半暗带SDF-1/CXCR4 m RNA及其蛋白的表达均于7d达到高峰,14,28 d表达逐渐下降。其中7,14 d同一时间点组间比较,HP-MSCs组、MSCs+QRHY组及HP+QRHY组二者的表达均明显优于N-MSCs组(P<0.01,P<0.05),而以HP+QRHY组增高最为明显(P<0.05,P<0.01),28 d后,各移植组的表达趋势趋同,但仍高于模型组(P<0.05)。结论缺氧预处理MSCs移植能够显著提高脑缺血再灌注损伤大鼠SDF-1/CXCR4的表达,清热化瘀方能够进一步上调SDF/1CXCR4的表达,减少细胞凋亡。
Objective To observe the effects of hypoxic preconditioning of bone marrow mesenchymal stem cells (MSCs) transplantation on the expression of SDF-1 / CXCR4 mRNA and protein after cerebral ischemia-reperfusion injury in rats and the intervention of clearing heat and removing stasis recipe. Methods Twenty-two SD rats were randomly divided into 6 groups: sham operation group (SO), model group (MCAO), MSCs transplantation control group (N MSCs transplantation, MSCs transplantation combined with MSCs + QRHY, HP-MSCs transplantation combined with HP + QRHY. Thirty-six rats in each group were divided into 3 subgroups and 12 rats in each subgroup according to the time points of 7,14,28 d. The expression changes of SDF-1 / CXCR4 m RNA and its protein at 3 time points were observed by q RT-PCR and Western blot. Apoptosis of neurons was detected by TUNEL method. Results The expression of SDF-1 / CXCR4 m RNA and its protein in ischemic penumbra of each group all reached a peak at 7d, and gradually decreased at 14 and 28 d. MSCs + QRHY group and HP + QRHY group were significantly better than the N-MSCs group (P <0.01, P <0.05) at 7 and 14 days, In HP + QRHY group, the increase was the most significant (P <0.05, P <0.01). After 28 days, the trend of expression in each group was similar but still higher than that in model group (P <0.05). Conclusion MSCs transplantation with hypoxic preconditioning can significantly increase the expression of SDF-1 / CXCR4 in rats with cerebral ischemia-reperfusion injury. Qingre Huayu prescription can further up-regulate the expression of SDF / 1CXCR4 and decrease apoptosis.