Expression of mir-199a in ulcerative colitis rats induced by 2, 4, 6-trinitrobenzene sulfonic acid/ethanol and study on the effect of tripteryginum wilfordii polyglycoside / 中国药学杂志

ABSTRACT

OBJECTIVE: To explore the expression of miR-199a in ulcerative colitis (UC) rats induced by 2, 4, 6-trinitrobenzene sulfonic (TNBS)/ethanol and the study on the effect of TWP on them. METHODS: Through injecting TNBS/ethyl alcohol acid liquid into the anus of the rats to establish the UC rat model. The colitics commom morphous damage and grade the histopathological score (CMDI) of colon mucosa injury were evaluated. Chip analysis and Real-time PCR were used to verify the expression of miR-199a in each colon mucosa tissue. Based on the expression profile, the downstream target genes mRNA in milwalk database was selected, then the expression of target genes mRNA by Real-time PCR in each group was veritied, at last the relevant signal pathway in the DAVID database was analyzed. Doing these to analyse the target gene mRNA regulated by the miR-199a in the inflammatory activity of UC. RESULTS: Compared with the model group, TWP high dose group was significantly lower on gross morphological damage score and histopathological injury score(P < 0.01). Chip analysis showed that in model group, the expression of miR-199a was significantly higher than the normal group(P < 0.01), and expression of the AZA group was significantly lower than the model group(P < 0.01, P < 0.05). The expression of miR-199a-3p in medium dose group and the expression of miR-199a-5p in high dose group were significantly lower than the model group(P < 0.05). The results of Real-time PCR showed that expression of miR-199a in the model group was significantly increased than that in the normal group(P < 0.01). The expression of miR-199a-3p in TWP medium dose group, high dose group and AZA group were decreased than that in model group(P < 0.05). Meanwhile, the expression of miR-199a-5p in TWP medium dose group was decreased than that in model group(P < 0.05). The gene expression profile showed that FASL was the target gene of miR-199a. In the model group, the expression of FASL was higher than that in the normal group. The expression of FASL in AZA group was significantly decreased than that in the model group(P < 0.01). The results by the Real-time PCR of the target gene FASL showed that in the model group, the expression of FASL was higher than that in the normal group (P < 0.01). The expression of FASL in medium dose group, high dose group and AZA group were significantly decreased than that in the model group (P < 0.01). CONCLUSION: miR-199a is up-regulated in TNBS/Ethanol UC rats, and FASL is the downstream target gene of miR-199a. TWP can reduce the UC's inflammatory effectively and decrease the up-regulated miR-199a in UC. FASL is up-regulated in UC's inflammatory activity. TWP can reduce downstream target gene FASL of miR-199a.