电针耳甲区对糖尿病大鼠脑微血管内皮细胞肿瘤坏死因子-α mRNA、细胞黏附因子-1和血管间黏附分子-1蛋白表达的影响  

Effects of Serum Derived from Rats Undergone Auricular Acupuncture Intervention on Expression of TNF-α mRNA,Cell Adhesion Factor-1 and Vascular Intercellular Adhesion Molecule-1 Proteins of Incubated Cerebral Microvascular Endotheliocytes with Diabetic In

作　　者：梅志刚[1] 曾永保[1] 王明智[1] 刘晓洁[1] 肖长义[1] 黎家华[1] 

机构地区：[1]三峡大学医学院,宜昌443002

出　　处：《针刺研究》2012年第6期440-446,共7页Acupuncture Research

基　　金：国家自然科学基金项目(30801486);三峡大学博士科研启动项目(0620070106)

摘　　要：目的:观察耳穴电针血清对糖尿病脑微血管内皮细胞损伤的保护作用,探讨耳针作用机制。方法:SD大鼠随机分为7组:正常组、模型组(高糖高脂进食,链脲佐菌素注射诱导)、耳针组(电针耳"胰胆""神门"穴)、迷走神经切断组(同侧迷走神经切断加耳针)、阿托品组、六烃季铵组和α-银环蛇毒素组(分别尾静脉注射阿托品、六烃季铵和α-银环蛇毒素外加耳针),10d后采集血清备用。正常培养的脑微血管内皮细胞分为同动物实验分组一致的7组,除正常组外,其它6组细胞建立糖尿病血管并发症细胞模型,48h后,收集细胞及条件培养液。观察各组细胞形态学和超微结构变化,检测各组细胞肿瘤坏死因子(TNF)-αmRNA表达情况,以及细胞条件培养液中细胞黏附分子-1(sICAM-1)和血管间黏附因子-1(sVCAM-1)的含量。结果:与模型组比较,耳针组、阿托品组细胞损伤较轻,细胞回缩数量明显减少,线粒体肿胀有改善,细胞TNF-αmRNA的表达、条件培养液sICAM-1和sVCAM-1的浓度显著下降(P<0.01,P<0.05);迷走神经切断组、六烃季铵组及α-银环蛇毒素组,耳针效应消失,细胞损伤与模型组相似,TNF-αmRNA表达以及sI-CAM-1和sVCAM-1蛋白浓度显著高于耳针组(P<0.01,P<0.05),而与模型组比较差异无统计学意义(P>0.05)。结论:耳针刺激具有抑制糖尿病血管并发症炎性反应,抑制细胞黏附等作用,上述抗炎作用可能是通过激活迷走神经,促进乙酰胆碱与其受体的α-亚单位结合而实现的。Objective To observe the protective effect of serum derived from rats undergone auricular electroacupunc- ture （EA） stimulation on the incubated cerebral microvascular endotheliocytes with diabetic injury so as to investigate the underl- ying mechanism of cholinergic anti-inflammatory action. Methods SD rats were randomized into normal group （n = 10）, diabe- tic model group （n = 6）, auricular EA group （n = 8）, vagotomy＋ EA group （n = 7, received ipsilateral vagotomy before auricular EA stimulation）, atropine＋ EA group （n = 8）, hexamethonium＋ EA group （n = 7） and a-bungaretoxin＋ EA group （n = 7）. Dia- betic mellitus model was established by feeding the rats with high sugar, high fat forage and intraperitoneal injection of 1% strep- tozotocin injection （STZ, 35 mg/kg）. EA was applied to ipsilateral ＂Yi-Dan＂-point and ＂Er-Shenmen＂ for 30 min, once daily for 10 days. Atropine（0. 1 mg/kg, an anticholinergic drug）, hexamethonium （10 mg/kg, an antagonist of the nicotinic acetylcholine receptors located in sympathetic and parasympathetic ganglia） and a-bungarotoxin （1.0 μg/kg, a type of a-neurotoxin that is known to bind irreversibly and competitively to the nicotinic acetylcholine receptors） were given to the rats by tail venous injec- tion, respectively, before ipsilateral auricular EA intervention, once daily for 10 days. Blood samples from rats of each group were then collected. Normally cultured rat brain micrevascular endotheliocytes were randomly divided into the same 7 groups. The diabetic-like damage model of cerebral microvascular endotheliocytes was established in the 6 groups except the normal oroup byadding the fluid containing glucose （20 mmol/L）, insulin （100 mU/L） and oxidized low density lipoprotein （200 mg/L） to the cul- ture medium. After 48 hours＇ incubation, the conditional culture solutions were collected for filtration and degerming. Morphologi- cal changes and cellular ultra-microstructure were examined using light micr

关 键 词：耳针糖尿病损伤 脑微血管内皮细胞胆碱能抗炎通路 

分 类 号：R245.323[医药卫生—针灸推拿学]