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目的观察针灸对溃疡性结肠炎大鼠IL1β和IL6mRNA表达的影响.方法采用人结肠术后新鲜结肠粘膜,加适量生理盐水制成粘膜匀浆,以3000r/min离心,取上清液,测定蛋白含量,并与Fruend佐剂混合配成乳剂.模型组大鼠首次足跖内注射抗原后,并于d10,d17,d24,d31分别于足跖、背部、腹股沟、腹腔内(末次注射不加佐剂)加强注射,至血清抗结肠抗体到一定效价后,再用20mL/L福尔马林和抗原液(不加佐剂)分别灌肠,建立溃疡性结肠炎大鼠模型.随机分为模型对照组(8只),电针组(8只),隔药灸组(8只).正常对照组大鼠8只.隔药灸组:用精制艾绒制成的艾炷在天枢(双)、气海穴位上隔药饼(药饼配方:附子、肉桂等药)灸2壮,1次/d,共14次;电针组:用间歇式脉冲波刺激天枢(双),气海,频率2Hz,强度4mA,时间20min,1次/d,共14次.治疗结束后,四组大鼠同时处死,分离脾脏、割取远端结肠.采用异硫氰酸胍/酚/氯仿一步法抽提组织总RNA,逆转录-聚合酶链反应(RTPCR)技术观察组织中IL1β,IL6mRNA的表达.结果正常对照组大鼠脾脏、结肠粘膜中未见IL1β,IL6mRNA的表达,模型对?
Objective To observe the effect of acupuncture on IL1β and IL6 mRNA expression in rats with ulcerative colitis. Methods The fresh colonic mucosa of human colon after operation and normal saline were added to make the mucosa homogenate. The supernatant was centrifuged at 3000 r / min. The protein content was determined and mixed with Fruend adjuvant to form the emulsion. After the first intraplantar injection of antigen in model group, the rats in model group were injected intensively on d10, d17, d24 and d31 in the plantar, dorsal, groin and intraperitoneal (the last injection without adjuvant), and the serum anti-colon antibody was effective After the price, and then 20mL / L formalin and antigen solution (without adjuvant) were enema, the establishment of ulcerative colitis rat model. Randomly divided into model control group (8), EA group (8), drug moxibustion group (8). Eight normal control rats. Moxibustion moxibustion group: made with refined moxa moxa moxibustion in Tianshu (double), gas sea points separated cake (cake recipe: aconite, cinnamon and other drugs) 2 strong, 1 / d, a total of 14 Electroacupuncture group: Tianshu (double), gas sea, frequency 2Hz, intensity 4mA, time 20min, 1 time / d were stimulated by intermittent pulse wave for 14 times. After treatment, four groups of rats were killed at the same time, the spleens were isolated and the distal colon was harvested. Total RNA was extracted by one-step method with guanidinium isothiocyanate / phenol / chloroform and the expression of IL-1β, IL-6 mRNA was observed by reverse transcription-polymerase chain reaction (RTPCR). Results The normal control group rats spleen, colonic mucosa did not see IL 1β, IL 6 mRNA expression, the model?