大豆异黄酮和法舒地尔联合应用对大鼠脑缺血再灌注损伤的保护作用
Protective effects of soybean isoflavones combined with fasudil on cerebral ischemia reperfusion injury in rats
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摘要: 目的：探讨大豆异黄酮（SI）和法舒地尔（Fas）对大鼠脑缺血再灌注后脑组织高迁移率族蛋白B1（HMGB1）、血管细胞黏附分子-1（VCAM-1）和核转录因子κB1（NF-κB1）表达的影响。方法：以体质量240~260 g雄性SD大鼠为实验对象，分为假手术组（S组）、缺血再灌注组（IR组）、IR+Fas组、IR+SI组和IR+Fas+SI组，HE染色法观察脑组织病理形态学改变，免疫组织化学法和Western blot法检测NF-κB1（p50）和HMGB1蛋白的定性和定量表达，荧光定量PCR测定脑组织HMGB1、NF-κB1和VCAM-1 mRNA的表达。结果：IR组脑神经细胞溶解性坏死，核固缩，组织间隙明显水肿增大；与IR组比较，各用药组脑组织损伤均有不同程度减轻，核固缩减少，组织间隙水肿改善。免疫组织化学法结果示，与S组比较，IR组NF-κB1和HMGB1的阳性细胞表达量均显著升高（P < 0.01）；与IR组比较，各用药组NF-κB1和HMGB1的阳性细胞数均明显降低（P < 0.01），其中IR+Fas+SI组与单独用药组比较降低均更显著（P < 0.01）。与S组比较，IR组HMGB1蛋白及HMGB1、NF-κB1和VCAM-1 mRNA表达均显著升高（P < 0.01）；与IR组比较，各用药组HMGB1蛋白及HMGB1、NF-κB1和VCAM-1 mRNA表达均明显降低（P < 0.01），其中IR+Fas+SI组与单独用药组比较下降均更显著（P < 0.01）。结论：SI和Fas对大鼠脑缺血再灌注损伤的保护作用可能是通过抑制HMGB1、NF-κB1和VCAM-1的表达，从而抑制NF-κB信号通路，以减弱炎症反应对脑组织的损伤，且联合应用比单独应用效果更显著。Abstract: Objective: To investigate the effects of soybean isoflavones(SI) combined with fasudil(Fas) on the expressions of high mobility group box 1(HMGB1),vascular cell adhesion molecule-1(VCAM-1),nuclear factor κB1(NF-κB1) in rats with cerebral ischemia reperfusion.Methods: The weight 240 to 260 g male SD rats were randomly divided into the sham group(S group),ischemia reperfusion(IR group),IR and Fas group,IR and SI group,and IR,Fas and SI group.The cerebral pathological change was observed by HE staining.The qualitative and quantitative expressions of NF-κB1(p50) and HMGB1 in cerebral tissue were measured by immunohistochemistry and western blotting,respectively.The expressions of HMGB1,NF-κB1 and VCAM-1 mRNA were examined by qRT-PCR.Results: Compared with the single drug group,the decreasing of the positive cells with NF-κB1 and HMGB1 expressions in IR,Fas and SI group were the more significant.Compared with the S group,the protein expression of HMGB1,and mRNA expression of HMGB1,NF-κB1and VCAM-1 in IR group increased significantly(P < 0.01).Compared with the IR group,the protein expression of HMGB1,and mRNA expressions of HMGB1,NF-κB1and VCAM-1 in other drug groups decreased significantly(P < 0.01).Compared with the single drug group,the decreasing of the protein expression of HMGB1,and mRNA expressions of HMGB1,NF-κB1 and VCAM-1 in IR,Fas and SI group were the more significant(P < 0.01).Conclusions: The protective effects of SI combined with Fas on cerebral ischemia reperfusion injury in rats maybe be implemented by inhibiting the expressions of HMGB1,NF-κB1 and VCAM-1,which can attenuate the signal channel of NF-κB,and decrease the cerebral tissue injury casued by inflammatory response.The protective effects of drug combination is significantly better than that of single drug.
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Key words:
- reperfusion injury /
- cerebral ischemia /
- soybean isoflavones /
- fasudil /
- nuclear factor κB /
- rats
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