脑出血各证型患者细胞自噬及Cathepsin D表达差异研究

Various Syndrome Autophagy of Intracerebral Hemorrhage and Expression Variation of Cathepsin

目的观察脑出血(ICH)闭脱证患者血肿周围组织细胞自噬和自噬相关蛋白Cathepsin D的表达差异。方法将27例ICH患者根据中医辨证分为阳闭证(12例)、阴闭证(9例)、脱证(6例)3组,评估出血量及格拉斯哥昏迷量表(GCS)评分、美国国立卫生研究院卒中量表(NIHSS)评分;另选6例颅内深部动静脉畸形(AVM)需开颅手术,但未发生脑出血的患者,作为对照组;采用透射电镜(TEM)技术和免疫组织化学法(IHC)检测细胞内自噬泡及Cathepsin D蛋白表达情况。结果 ICH患者血肿周围脑组织中自噬泡数量及Cathepsin D表达明显高于对照组(P<0.01)。各证型按自噬泡数量、Cathepsin D表达水平、出血量、NIHSS评分排列由高至低均为:脱证、阴闭证、阳闭证(P<0.01,P<0.05);按GCS评分排列由高至低为阳闭证、阴闭证、脱证(P<0.01)。自噬泡数量、Cathepsin D表达与出血量、NIHSS评分呈正相关(P<0.01);与GCS评分呈负相关(P<0.01)。结论细胞自噬参与了ICH后继发性神经细胞损伤,闭脱证患者血肿周围组织细胞自噬和自噬相关蛋白Cathepsin D的表达存在差异。

Objective To observe the differences of neural autophagy and the expression of Ca- thepsin D in patients with excess syndrome of stroke and depletion syndrome of stroke in the peri-hema- tomal brain tissue of intracerebral hemorrhage（ICH）. Methods Twenty-seven patients were assigned to three groups according to syndrome typing,i.e,as Yang excess group（12 cases） ,Yin excess group（9 cases） ,and depletion syndrome group（6 cases）. The hemorrhage volume was assessed. National Insti- tute of Health Stroke Scale（NIHSS） and Glasgow Coma Scale（GCS） scores were recorded. Another six encephalic arteriovenous malformation （AVM） patients that require craniotomy, without occurrence of ICH, were enrolled as the control group. Autophagy in the peri-hematomal brain tissue was observed by using transmission electron microscopy（TEM）.The expressions of Cathepsin D was detected by immuno- histochemistry（IHC）. Results The number of autophagic vacuoles and the expression of Cathepsin D were significantly increased in the peri-hematomal brain tissue of ICH patients compared with that in the control group（P 〈0.01 ）. In accordance with autophagic amount, Cathepsin D expression level, bleeding amount, and NIHSS, syndromes types were sequenced from high to low as depletion syndrome, Yin ex- cess syndrome and Yang excess syndrome （P 〈0.01, P 〈0.05） ~ in accordance with GCS, syndromes types were sequenced as Yang excess syndrome, Yin excess syndrome and depletion syndrome （P 〈 0.01 ）. Autophagic amount and Cathepsin D expression were positively correlated to bleeding volume and NIHSS （P 〈0.01 ）, and negatively correlated to GCS （P 〈0.01 ）.Conclusions Autophagy participates in secondary neuron injury after ICH. Autophagy and the expression of Cathepsin D were different in the pe-ri-hematomal brain tissue of patients with excess syndrome of stroke and depletion syndrome of stroke.