【摘要】 背景:急性脑缺血时基质金属蛋白酶通过降解细胞外基质,破坏内皮细胞间的紧密连接而增加微血管的通透性,破坏血脑屏障,导致脑水肿的发生。目的:旨在从基质金属蛋白酶及细胞外基质方面,探讨补阳还五汤联合骨髓间充质干细胞移植对脑缺血再灌注大鼠的治疗作用机制。方法:将96只SD大鼠随机分为模型组,组织型基质金属蛋白酶抑制剂1组,组织型基质金属蛋白酶抑制剂1+骨髓间充质干细胞组(简称骨髓间充质干细胞组),组织型基质金属蛋白酶抑制剂1+补阳还五汤联合骨髓间充质干细胞组(简称联合组,分12,24,36,48 h 4个亚组)。构建大鼠脑缺血再灌注模型,在立体定位仪下采用微量进样器脑部给予基质金属酶抑制剂及培养的骨髓间充质干细胞;联合组术前7 d开始给予补阳还五汤(10 m L/kg)灌胃,其余各组给予等剂量的生理盐水。给药10 d后采集实验动物血清和脑组织,采用ELISA法测定血清中基质金属蛋白酶2、基质金属蛋白酶9水平,明胶酶谱法分析脑组织基质金属蛋白酶9活性。结果与结论:与模型组相比,各组血清基质金属蛋白酶2、基质金属蛋白酶9水平和脑组织基质金属蛋白酶9的活性均有不同程度的下降,其中基质金属蛋白酶9下降明显(P<0.05);与骨髓间充质干细胞组相比,联合组36 h和联合组48 h血清基质金属蛋白酶2、基质金属蛋白酶9水平和脑组织基质金属蛋白酶9、基质金属蛋白酶9前体活性下降最为明显(P<0.01)。结果表明补阳还五汤联合骨髓间充质干细胞可与组织型基质金属蛋白酶抑制剂1协同作用,从而抑制因基质金属蛋白酶2、基质金属蛋白酶9所致的细胞外基质降解,修复受损的血脑屏障及防治缺血后脑水肿的形成。更多还原

【Abstract】 BACKGROUND: When acute cerebral ischemia attacks, matrix metalloproteinases(MMPs) lead to the occurrence of cerebral edema through degrading the extracellular matrix, breaking the close connection between endothelial cells, increasing the permeability of capillaries, and destroying the blood brain barrier. OBJECTIVE: From the aspects of MMPs and extracellular matrix, to discuss the therapeutic effects of buyang huanwu decoction combined with bone marrow mesenchymal stem cell(BMSCs) transplantation on cerebral ischemia-reperfusion injury.METHODS: A total of 96 Sprague-Dawley rats were randomly divided into model group, tissue inhibitor of MMPs-1(TIMP-1) group, TIMP-1+BMSCs group(BMSCs group) and buyang huanwu decoction+BMSCs+TIMP-1 group(combined group that was divided into four subgroups, 12-, 24-, 36-, 48-hour groups). Rat models of cerebral ischemia-reperfusion were constructed, and TIMP-1 and BMSCs were injected to the brain of rats by a microinjector in a stereotaxic apparatus. Rats in the combined group were given buyang huanwu decoction(10 m L/kg), and rats in the other groups were given the same volume of normal saline at 7 days before surgery. After 10 days of administration, serum samples and brain tissues were collected to determine MMP-2 and MMP-9 levels using ELISA and to detect MMP-9 activity using gelatinases spectrometry method. RESULTS AND CONCLUSION: Compared with the model group, the levels of MMP-2 and MMP-9 in the serum and MMP-9 activity in the brain were decreased in the other groups to different extents, especially the levels of MMP-9(P < 0.05). Compared with the BMSCs group, the levels of MMP-2 and MMP-9 in serum as well as activities of MMP-9 and pro-MMP-9 in the brain were decreased significantly in the combined group at 36 and 48 hours after treatment(P < 0.01). The results show that the buyang huanwu decoction can be mutually cooperated with TIMP-1 to inhibit the degradation of extracellular matrix induced by MMP-2 and MMP- 9, repair the damaged blood brain barrier, prevent and cure cerebral edema after ischemia.更多还原