Study On The Regulation Of Radix Pulsatillae Alcohol Extract In Contols Matrix Metalloproteinase-3 Expression In Colonic Mucosa Of Rat With Trinitrobenzenesulfonic Acid Induced Colitis

Objective: To explore the anti-inflammation of radix pulsatillae alcohol extract on trinitrobenzenesulfonic acid(TNBS) induced rat colitis, to observe the effects of the radix pulsatillae alcohol extract on the gene expression of Matrix Metalloproteinase-3(MMP3), so as to interpret further the mechanism that radix pulsatillae alcohol extract cures rat colitis. Methods: 28 male and female rats were randomized into 3 groups: radix pulsatillae alcohol extract treated group(Group A,n=10), TNBS controlled group(Goup B,n=10) and normal controlled group(Group C,n=8). All rats were on an absolute diety for 24 hours except drinking water freely, then 1ml enema containing TNBS (100mg/kg) dissolved in 50% ethanol was instilled into the colon about 8cm away from anus in Group A and Group B with a silica gel tube, the Group C rats were instilled with sodium chloride in an equal volume as Group A, keeping the anus in superior position for 2～3 minutes to avoid that enema reflows. The Group A rats were perfused with 2ml radix pulsatillae alcohol extract at the dosage of 2g/kg/d for 7 days; the Group B and Group C rats were treated with distilled sodium chloride, all rats drank water freely. Mental status, eating, activity, stools, body weights were observed and were cut the neck off on the 8th day, segregated and taked down the colon, and then cleaned the enteric cavity soil with cotton bud. The appearance of hyperemia, the size and distribution of ulcers, the bleeding, the disease acticity index(DAI) and the score of the tissue damages of colonic mucosa were calculated together with HE staining. The levels of tumor necrosis factor-a(TNF-α) and interleukin-10(IL-10) in colonic tissue were defected by enzyme linked immune sorbent assay (ELISA). The expression of NF-κB p65 in colonic tissue was observed by immunohistochemistry. The expression of MMP3-mRNA was detected by realtime fluorescent quantitation polymerase chain reaction (RT-PCR). Results: Compared with Goup B, the disease acticity index(DAI) and histological damages of colonic mucosa in Group A were significantly lowered(P<0.01); the levels of TNF-αwere significantly decreased(P<0.05), but that of IL-10 were significantly increased(P<0.01); the expression of NF-κBp65 were significantly decreased (P<0.05); the expression of MMP3-mRNA in colonic tissue were significantly decreased(P<0.01); the expression of MMP3-mRNA has a significant positive correlation with those of NF-KB p65 (r=0.677, P<0.01). Conclusion: According to the research, radix palstillae alcohol extract has an exact anti-inflammatory effects on TNBS induced rat colitis. The mechanism is possibly related to down-regulate the expression of MMP3-mRNA, inhibit the activation of NF-κB, regulate network balance of proinflamtory cytokines and anti-inflammatory cytokines, then reduces the degraded and destructive processes of MMP3 in rats colon tissue, improves the tissue immune function and reduces the infiltration of inflammatory cell.