Study of the mechanisms of acupuncture and moxibustion treatment for ulcerative colitis rats in view of the gene expression of cytokines

摘要

AIM: To observe the effect of acupuncture and moxibustion on the expression of IL-1β and IL-6 mRNA in ulcerative colitis rats.METHODS: The SD rat ulcerative colitis model was created by immunological method associated with local stimulation. Colonic mucosa was prepared from human fresh surgical colonic specimens, homogenized by adding appropriate amount of normal saline and centrifuged at 3000 r/min. The supernatant was collected for measurement of protein conentration and then mixed with Freund adjuvant. This antigen fluid was first injected into the plantae of the model group rats, and then into their plantae, dorsa, inguina and abdominal cavities (no Freund adjuvant for the last injection) again on the 10th, 17th, 24th and 31st day. When a certain titer of serum anti-colonic anti body was reached, 2% formalin and antigen fluid ( no Freund adjuvant ) were administered separately by enema. The ulcerative colitis rat model was thus set up. The animals were randomly divided into four groups: model control group ( MC, n = 8 ), electro-acupuncture group (EA, n = 8), herbs-partition moxibustion group (HPM 8), normal control group ( NC, n = 8 ). HPM: Moxa cones made of refined mugwort floss were placed on the medicinal pad (medicinal pad dispensing: Radix Aconiti praeparata, cortex Cinnamomi, etc) for Qihai (RN 6) and Tianshu (S T 25, bilateral) and ignited. Two moxa cones were used for each acupoint once a day and 14 times in all. EA: Tianshu (bilateral) and Qihai were stimulated by the intermittent pulse with 2 Hz frequency, 4mA intensity for 20 min once a day and 14 times in all. After treatment, rats of all four groups 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 study the expression of IL-1β and IL-6 mRNA.RESULTS: IL-1β and IL-6 mRNAs were not detected in the spleen and colonic mucosa of the NC rats, whereas they were significantly expressed in that of the MC rats. IL-1β and IL-6 mRNAs we re markedly lower in the EA and HPM rats than that in MC rats. There was no significant difference between the levels of IL-1β and IL-6 mRNAs in the EA and HPM rats. The expressions of IL-1β and IL-6 mRNAs were nearly the same in the spleen and colon of all groups.CONCLUSION: Acupuncture and moxibustion greatly inhibited the expression of IL-1β and IL-6 mRNA in the experimental ulcerative colitis rats.