栝楼桂枝颗粒对脑缺血再灌注损伤大鼠线粒体能量代谢的影响

Effects of Gualou Guizhi Granules on energy metabolism of mitochondria following cerebral ischemia reperfusion injury rats

目的:探讨栝楼桂枝颗粒是否通过调控线粒体能量代谢对脑缺血再灌注损伤大鼠起到脑保护作用。方法:SD大鼠随机分为假手术组、模型组和中药组。采用线栓法制备脑缺血再灌注模型,采用神经功能缺损评分法(mNSS)进行大鼠神经功能评分,测定大鼠脑梗死面积,采用透射电镜观察脑线粒体超微结构的改变;采用分光光度法测定线粒体呼吸链酶复合物Ⅰ的活力来评价栝楼桂枝颗粒对模型大鼠线粒体能量代谢的影响;利用免疫组化法观察解偶联蛋白4(UCP4)和ATPase 6在脑组织中的表达。结果:与模型组比较,中药组大鼠神经行为学情况得到显著改善(P<0.05),缺损程度降低(P<0.05);中药组对模型大鼠脑线粒体超微结构病理状态有改善作用;中药组能够提高模型大鼠脑组织中线粒体呼吸链酶复合物Ⅰ的活力(P<0.05)、脑组织中ATP的含量(P<0.05),显著提高模型大鼠脑组织中Na+-K+-ATPase、Ca2+-ATPase、Mg2+-ATPase、Ca2+-Mg2+-ATPase的活性(P<0.05,P<0.01);中药组能够恢复模型大鼠脑组织中UCP4和ATPase 6的表达(P<0.01)。结论:栝楼桂枝颗粒可以通过保护线粒体而调节线粒体能量代谢从而起神经保护作用。

Objective: To investigate the effects of Gualou Guizhi Granules on mitochondria of energy metabolism following cerebral ischemia reperfusion injury. Methods: A rat model of cerebral ischemia reperfusion injury was established by middle cerebral artery occlusion, SD rats were divided into sham-operation group, model group and Chinese medicine group, using modified neurological severity scores(mNSS) to evaluate the neural function and the area of cerebral infarction was determined;Transmission electron microscopy was utilized to analyze the ultrastructural changes of mitochondria;Evaluated the influence of Gualou Guizhi Granules on the activity of mitochondrial respiratory chain complexes Ⅰ detected by UV-vis spectrophotometry, mitochondrial energy metabolism of model rats. Immunocytochemical staining methods were used to observe the expression of UCP4 and ATPase 6 in brain issue of rats. Results: Compared with model group, the nerve function defect and cerebral infarction area score of the Chinese medicine group increased significantly(P<0.05);Chinese medicine group could improve the ultrastructural pathological state of mitochondria(P<0.05);Chinese medicine group could increase the activity of mitochondrial complex Ⅰ(P<0.05), the ATP content in cerebral tissue(P<0.05), and could significantly increase the activity of Na+-K+-ATPase, Ca2+-ATPase, Mg2+-ATPase, Ca2+-Mg2+-ATPase(P<0.05, P<0.01);Chinese medicine group could restore the expression of UCP4 and ATPase 6 in brain issue of rats(P<0.01). Conclusion: Gualou Guizhi Granules has neuroprotective effect by protecting mitochondria and adjusting the mitochondrial energy metabolism.