电针对溃疡性结肠炎大鼠结肠细胞自噬的影响  

Effect of electroacupuncture on autophagy of colonic cells in rats with ulcerative colitis

作　　者：徐慧超 王海军 曹玉霞 武荣林 蒋子文 李让钱 冀来喜 XU Hui-chao;WANG Hai-jun;CAO Yu-xia;WU Rong-lin;JIANG Zi-wen;LI Rang-qian;JI Lai-xi(Second Clinical College,Shanxi University of Traditional Chinese Medicine,Jinzhong 030619,China;Affiliated Acupuncture and Massage Hospital of Shanxi University of Traditional Chinese Medicine,Taiyuan 030006,China)

机构地区：[1]山西中医药大学第二临床学院,山西晋中030619 [2]山西中医药大学附属针灸推拿医院,山西太原030006

出　　处：《时珍国医国药》2024年第4期1022-1024,F0003,F0004,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(82074549)。

摘　　要：目的探讨电针“肠病方”对溃疡性结肠炎(UC)大鼠结肠细胞自噬水平的调控作用。方法将40只雄性SD大鼠随机分为5组:正常对照组(正常组),UC模型组(模型组),电针治疗组(电针组),雷帕霉素(RAPA)溶液干预组(RAPA组),3-甲基腺嘌呤(3-MA)溶液干预组(3-MA组),每组8只大鼠。正常组大鼠给予正常饮食,其余4组大鼠采用5%葡聚糖硫酸钠(DSS)溶液建立UC大鼠模型。造模成功后,电针组大鼠采用电针刺激大鼠“中脘”及双侧“天枢”、“上巨虚”五个穴位,疏密波,频率10 Hz/50 Hz,强度以大鼠肢体轻微颤动,电针20 min。RAPA组大鼠腹腔注射2 mg/kg RAPA溶液,3-MA组大鼠腹腔注射1.5 mg/100 g 3-MA溶液。测量大鼠体质量、结肠长度,计算疾病活动指数(DAI)评分,并取各组大鼠血清及结肠组织,通过HE染色观察大鼠结肠形态学变化;酶联免疫吸附实验(ELISA)检测血清中肿瘤坏死因子(TNF-α)、白细胞介素(IL)-6、IL-2、IL-10含量;超微电镜观察大鼠结肠黏膜上皮细胞超微结构;Western blot法检测自噬相关蛋白(ATG)5、ATG 12、LC 3和p 62蛋白相对含量;实时荧光定量PCR(qPCR)检测ATG 5、ATG 12、LC 3和p 62 mRNA含量。结果与正常组相比,模型组、3-MA组大鼠DAI评分分值增加,结肠长度缩短,血清中IL-2、IL-10水平降低,TNF-α、IL-6水平增加,结肠组织中ATG 5、ATG 12、LC 3相对蛋白表达含量及该基因mRNA的相对含量显著降低,p 62相对蛋白表达含量及其基因mRNA的相对含量显著升高(P<0.05,P<0.01);与模型组相比,电针组和RAPA组大鼠DAI评分分值下降,结肠长度增加,血清中IL-2、IL-10水平升高,TNF-α、IL-6水平降低,结肠组织中ATG 5、ATG 12、LC 3相对蛋白表达含量及该基因mRNA的相对含量显著升高,p 62相对蛋白表达含量及其基因mRNA的相对含量显著降低(P<0.05,P<0.01);与电针组相比,3-MA组大鼠DAI评分分值增加,结肠长度缩短,血清中IL-2、IL-10水平降低,TNF-α、IL-6水平升Objective To investigate the effect of electroacupuncture(EA)of"Changbing Decoction"on the level of autophagy in colonic cells in rats with ulcerative colitis(UC).Methods The UC model was established by using 5%dextran sulfate sodium(DSS)solution.Forty male SD rats were randomly divided into 5 groups:Rats in the normal control group(normal group),UC model group(model group),EA treatment group(EA group),UC group(RAPA group),and UC group(3-MA group)were treated with intraperitoneal injection of rapamycin(RAPA group),and UC group(3-MA group)were treated with intraperitoneal injection of 3-methyladenine(3-MA group).Ea was applied to"Zhongwan"(CV 12),bilateral"Tianshu"(ST 25)and"Shangjuxu"(ST 37)for 20 min,with disperse-dense wave,10 Hz/50 Hz in frequency and slight tremor in the limbs of rats.The body weight and colon length of rats were measured,and the disease activity index(DAI)score was calculated.The serum and colon tissue of rats in each group were collected,and the morphological changes of colon were observed by HE staining.The levels of tumor necrosis factor(TNF)-α,interleukin(IL)-6,IL-2 and IL-10 in serum were detected by enzyme-linked immunosorbent assay(ELISA).The ultrastructure of colonic mucosal epithelial cells was observed by ultramicroelectron microscope.Western blot was used to detect the relative levels of autophagy-related proteins(ATG 5,ATG 12,LC 3 and p 62).Quantitative real-time PCR(qPCR)was used to detect the mRNA levels of ATG 5,ATG 12,LC 3 and p 62.The UC model was established by using 5%dextran sulfate sodium(DSS)solution.Forty male SD rats were randomly divided into 5 groups:Rats in the normal control group(normal group),UC model group(model group),EA treatment group(EA group),UC group(RAPA group),and UC group(3-MA group)were treated with intraperitoneal injection of rapamycin(RAPA group),and UC group(3-MA group)were treated with intraperitoneal injection of 3-methyladenine(3-MA group).Ea was applied to"Zhongwan"(CV 12),bilateral"Tianshu"(ST 25)and"Shangjuxu"(ST 37)for 20 min,with disperse

关 键 词：溃疡性结肠炎 电针 细胞自噬 

分 类 号：R2-03[医药卫生—中医学]