摘要
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?
AIM To observe the effect of acupuncture and moxibustion on the expression of IL 1β and IL 6 mRNA in ulcerative colitis rats. METHODS Human fresh surgical colonic mucosa was homogenized by adding appropriate amount of normal saline and centrifuged at 3000r/min . Protein content of the supernatant was measured and then mixed with Freund adjuvant, and injected into the plantae of the model group rats, then into the plantea, dorsa, inguen and abdominal cavities (no Freund adjuvant for the last injection) on the 10th, 17th, 24th and 31st day respectively. When a certain titer of serum anti colonic antibody was reached, 20mL/L formalin and antigen fluid (no Frenud adjuvant) were administered by enema to set up ulcerative colitis rat model. The animals were randomly divided into four groups: model control group (MC=8), electropuncture group (EP=8), herb spartition moxibustion group (HPM=8) and normal control group (NC=8). HPM: Mosa cones made of refined mugwort floss were placed on the medicinal pads (medicinal pad dispensing: Radix Aconiti praeparata, cortex Cinnamomi , etc) for Qihai (RN 6) and Tianshu (ST 25, bilateral) and ignited. Two moxa cones were used for each acupuncture once a day and 14 times in all. EP: Tianshu (bilateral) and Qihai were stimulated by the intermittent pulse with 2Hz frequency, 4mA intensity for 20 minutes once a day and 14 times in all. After treatment, all rats were killed simultaneously. The spleen was separated and the distal colon was dissected. Total tissue RNA was isolated by the guanidinium thiocyanate phenol chloroform extraction method. RT PCR technique was used to observe the expression of IL 1β and IL 6 mRNA. RESULTS IL 1β and IL 6 mRNA was not detected in the spleen and colonic mucosa of NC rats, while it was significantly expressed in that of MC rats. IL 1β and IL 6 mRNA was markedly lower in EP and HPM rats than that in MC rats. There was no significant difference in the levels of IL 1β and IL 6 mRNA between EP and HPW rats. The amount of
基金
国家自然科学基金
关键词
溃疡性结肠炎
针灸疗法
细胞因子
基因表达
colitis, ulcerative/therapy
acupuncture and moxibustion
cytokines
gene expression
RNA, messenger
