电针刺激对HGF-MSC减轻大鼠肺纤维化的影响  

Effect of electroacupuncture stimulation on HGF-MSC-induced reduction of pulmonary fibrosis in rats

作　　者：雷立华[1] 刘日烽[2] 郭依清 余剑波 林健清[2] 何诗灵 林小华 林君 郭廷生[2] 林群[2] Lei Lihua;Liu Rifeng;Guo Yiqing;Yu Jianbo;Lin Jianqing;He Shiling;Lin Xiaohua;Lin Jun;Guo Tingsheng;Lin Qun(Department of Anesthesiology,Fujian Provincial Hospital,Shengli Clinical Medical College of Fujian Medical University,Fuzhou 350001,China;Department of Anesthesiology,First Affiliated Hospital of Fujian Medical University,Fuzhou 350004,China;Department of Anesthesiology,Tianjin Nankai Hospital,Tianjin 300100,China)

机构地区：[1]福建省立医院麻醉二科福建医科大学省立临床学院,福州350001 [2]福建医科大学附属第一医院麻醉科,福州350005 [3]天津南开医院麻醉科,300100

出　　处：《中华麻醉学杂志》2019年第12期1509-1513,共5页Chinese Journal of Anesthesiology

基　　金：福建省医学创新课题(2016-CX-2,2016-CX-30);福建省自然科学基金面上项目(2019J0118)。

摘　　要：目的评价电针刺激对人肝细胞生长因子基因修饰骨髓间充质干细胞(HGF-MSC)减轻大鼠肺纤维化的影响。方法清洁级健康雄性F344大鼠40只,6周龄,体重200~250 g,采用随机数字表法分为5组(n=8):对照组(C组)、肺纤维化组(L组)、肺纤维化+HGF-MSC组(LH组)、肺纤维化+电针刺激组(LE组)和肺纤维化+电针刺激+HGF-MSC组(LEH组)。采用雾化吸入博来霉素5 mg/kg的方法制备大鼠肺纤维化模型。LH组和LEH组大鼠于吸入博来霉素后第15天尾静脉注射1 ml HGF-MSC悬浮液(106个/ml),L组注射等量生理盐水,LE组和LEH组,选取双侧足三里和肺俞穴进行电针刺激(疏密波,频率2/15 Hz,刺激强度以肢体出现轻微颤动为宜),30 min/d,持续28 d。博来霉素或者生理盐水吸入后第28天处死大鼠取肺组织,检测MDA、羟脯氨酸(HYP)、人HGF蛋白、SOD、谷胱甘肽过氧化物酶(GSH-Px)及核蛋白Nrf2表达的水平,根据Ashcroft评分评估肺纤维化程度。结果与C组比较,L组肺组织SOD和GSH-Px活性降低,羟脯氨酸和MDA含量、Ashcroft评分升高,核蛋白Nrf2表达上调(P<0.05);与L组比较,LE组和LH组肺组织SOD活性升高,MDA含量和Ashcroft评分降低,LH组羟脯氨酸含量降低,核蛋白Nrf2表达上调(P<0.05);与LEH组比较,LE组和LH组肺组织SOD活性降低,核蛋白Nrf2表达下调,MDA含量和Ashcroft评分升高(P<0.05)。结论电针刺激可增强HGF-MSC减轻大鼠肺纤维化的作用,其机制可能与上调Nrf2表达,增强抗氧化能力有关。Objective To evaluate the effect of electroacupuncture(EA)stimulation on human HGF gene modified bone marrow mesenchymal stem cells(HGF-MSC)-induced reduction of pulmonary fibrosis in rats.Methods Forty clean-grade healthy male F344 rats,aged 6 weeks,weighing 200-250 g,were divided into 5 groups(n=8 each)using a random number table method:control group(group C),pulmonary fibrosis group(group L),pulmonary fibrosis plus HGF-MSC group(group LH),pulmonary fibrosis+EA stimulation group(group LE),and pulmonary fibrosis+HGF-MSC+EA stimulation group(group LEH).Pulmonary fibrosis was induced by atomized inhalation of bleomycin 5 mg/kg in rats.HGF-MSC suspension(106 cells/ml)1 ml was injected via the the tail vein on 15th day after inhaling bleomycin in LH and LEH groups,while the equal volume of normal saline was given instead in group L.Bilateral acupoints Zusanli and Feishu were stimulated with disperse-dense waves(frequency 2/15 Hz,appropriate intensity of stimulation was slight fibrillation)for 30 min per day for 28 consecutive days in LE and LEH groups.The rats were sacrificed at 28 days after bleomycin or normal saline inhalation,and lung tissues were removed for determination of contents of malondialdehyde(MDA),hydroxyproline(HYP),human HGF protein,activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),and expression of Nrf2 in nucleoprotein and for microscopic examination.The degree of pulmonary fibrosis was assessed according to the Ashcroft score.Results Compared with group C,the activities of SOD and GSH-Px in lung tissues were significantly decreased,the contents of hydroxyproline and MDA and Ashcroft score were increased,and the expression of Nrf2 in nucleoprotein was up-regulated in group L(P<0.05).Compared with group L,the SOD activity in lung tissues was significantly increased,and the MDA content and Ashcroft score were decreased in LE and LH groups,and the hydroxyproline content was significantly decreased,and the expression of Nrf2 in nucleoprotein was up-regulated in group LH(P<0.05)

关 键 词：电针 肝细胞生长因子 间充质基质细胞 肺纤维化 

分 类 号：R24[医药卫生—中医临床基础]