| Taurochenodeoxycholic acid?TCDCA?is one of the major active components of bile acid.It shows the effect of not only inhibition of both acute and chronic inflammation but also regulation of the immune function.In order to further reveal the anti-inflammatory and immunomodulatory effects of TCDCA,the following researches were conducted in this study:?1?The time-effect curves were detected by real-time PCR for acting of TCDCA on fibroblast-like synoviocytes?FLS?of adjuvant arthritis model rats?AA rats?.?2?The effects of TCDCA on FLS of AA rats was detected using RNA-seq.?3?Differential expression genes GAPDH,GPX3,SRSF9,CTGF and CSTB were confirmed by real-time PCR.?4?The expression of GRs and the nuclear translocation effected by different concentrations of TCDCA in NR8383 cells were detected by immunofluorescence technique.?5?The effect of TCDCA on the gene expression levels of VCAM-1 and COX-2 in FLS of AA rats were evaluated by real-time PCR.?6?The effect of TCDCA on the expression of VCAM-1 in FLS of AA rats was also measured by ELISA.The results showed that the optimum treated time of TCDCA on FLS was 12 h.Twelve samples from four experimental groups were sequenced using RNA-seq technology and 24 differential genes were screened.There were 5 differential genes related to anti-inflammatory immunity,identified GAPDH,GPX3,SRSF9,CTGF and CSTB;TCDCA could upregulate the m RNA expression levels of SRSF9,GPX3 and CSTB.The results also demonstrated that TCDCA at the concentration of 1×10-55 mol/L could significantly?p<0.05?upregulated the mRNA expression level of CTGF;while 10-5and 10-77 mol/L of TDCCA could significantly?p<0.01?up-regulated the GAPDH mRNA expression level;Moreover,TCDCA could significantly?p<0.01?activate GR receptor in NR8383 cells,and after adding RU486,a specific inhibitor of GR,the fluorescence value of GR receptor decreased significantly;While there was no significant difference between DEX group and TCDCA group?p>0.05?;TCDCA could inhibit the expression of VCAM-1 and COX-2 in FLS of AA rats.The main conclusions of this experimental study are as following:?1?The optimum time of TCDCA promoting TGR5 and GR receptor expression in FLS of AA rats was 12 h;?2?After TCDCA acted on FLS,there were altogether 24 differential genes were screened by RNA-seq,among which GAPDH,GPX3,SRSF9,CTGF and CSTB were associated with anti-inflammatory property and immunity regulation;?3?Validated by real-time PCR,GPX3 and SRSF9 are consistent with the RNA sequencing results,while CTGF,GAPDH and RNA sequencing results are not;CSTB was not identical to the RNA sequencing results but was in agreement with the literature reports;?4?TCDCA can upregulate the expression of GPX3,SRSF9 and CSTB in FLS of AA rats;?5?TCDCA can cause nuclear translocation of GR receptor in NR8383,and significantly inhibit the expression of VCAM-1 and COX-2 in FLS of AA rats.In summary,the anti-inflammatory and immunomodulatory effects of TCDCA are related to its up-regulation of GPX3,SRSF9,and CSTB expression levels and the activation of glucocorticoid receptors,followed by the inhibition of VCAM-1 and COX-2in FLS of AA rats. |
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