Preliminary Study Of Shexiang Wulong Pill On The Mechanism Of Regulation Of MiR-155 And T Cell Subsets Differentiation In The Treatment Of RA

Objectives To explore the effect of serum of Shexiang Wulong Pill on the expression level of miR-155 and the differentiation of T lymphocyte subsets in peripheral blood mononuclear cells(PBMCs)in RA patients.And to explore the correlationn between miR-155 and the differentiation of T lymphocyte subsets in RA,to provide a new theoretical basis for treatment of RA.Methods 1 The peripheral blood mononuclear cells(PBMCs)were collected from 30 RA patients who meet the criterion and 30 normal subjects.Flow cytometry was used to detect the relative percentage of CD4+T lymphocytes and CD8+T lymphocytes in PBMCs.CD4+/CD8+ was calculated.Then compare them in RA patients and normal subjects.2Each RA PBMCs were divided in Shexiang Wulong pill group,TGP group,normal saline group and control group,then the corresponding 1640 serum was given.3 48 hours later,flow cytometry was used to detect the relative percentage of CD4+T cells and CD8+T cells in each group of PBMCs.CD4+/CD8+ was calculated.Then compare them in each group.4 The expression of miR-155 was detected by real-time fluorescent quantitative PCR.Compare the differences in these groups.5 After the intervention of Shexiang Wulong pill,the relevance between the expression of CD4+T lymphocyte and miR-155 was analyzed.Preliminary judge on whether there is relevance between them.The data were input by EXCEL and analyzed by SPSS17.0.Independent sample T test was used between two groups.One-way ANOVA was used in groups.The Pearson correlation analysis was used to analyze the two variables of bivariate normal distribution.The difference of P<0.05 was statistically significant.Results 1 Compared with normal,there was no significant difference in CD8+T lymphocyte relative contents.But CD4+T lymphocyte relative percentage and CD4+/CD8+ increased more significantly than normal(P<0.05).2 After 48 hours,CD4+T lymphocyte relative percent content and CD4+/CD8+ were significantly lower than before in Shexiang Wulong pill group and TGP group(P<0.001<0.05).Compared with the normal saline group and blank serum group,CD4+T lymphocyte relative percentage and CD4+/CD8+ in Shexiang Wulong pill group and TGP group was decreased significantly(P<0.05).CD4+T lymphocyte relative percentage and CD4+/CD8+ in Shexiang Wulong pill group decreased significantly than TGP group(P=0.006<0.05,P=0.028<0.05).3 After48 hours,the expression of miR-155 in Shexiang Wulong pill group and TGP group were significantly decreased than the normal saline group and blank serum group(P<0.001).The expression of miR-155 in Shexiang Wulong pill group were significantly lower than that of TGP group(P<0.001).4 There was a positive correlation between the reduced expression of CD4+T lymphocytes and the down regulated expression of miR-155 in the two groups(r=0.38),which was statistically significant(P<0.05)Conclusions Abnormal differentiation of CD4+ T lymphocyte subsets in peripheral blood of patients with RA is the main phenomenon.Shexiang Wulong pill may reduce the expression of CD4+T lymphocytes and CD4+/CD8+ in peripheral blood of patients with RA.Shexiang Wulong pill can reduce the expression of miR-155 in PBMCs of RA patients and reduce the inflammatory reaction.There was a positive correlation between the down regulated expression of miR-155 and the decrease of CD4+T lymphocyte differentiation,which may be related to the mechanism of Shexiang Wulong pill reducing the inflammatory reaction.