ABSTRACT
Objective:To observe the Toll-like receptor 4(TLR4)and its negative regulating factorInterleukin-1 receptor-associated kinase-M(IRAK-M)in colonic mucosa of rats with experimental ulcerative colitis(UC), and to discuss the mechanism of the Chinese medicine Sishenwan. Method:The 90 Wistar rats were randomly divide into six groups, blank group, model group, sulfasalazine group(0.36 g·kg-1), Sishenwan low, medium and high-dose group(2.5, 5, 10 g·kg-1), 15 cases in each group. A rat model of UC was prepared by using a solution of trinitrobenzenesulfonic acid/ethano.The histopathological changes of colon were observed by hematoxylin-eosin (HE) staining. The contents of serum free triiodothyroid acid (FT3), serum free thyroxine (FT4), immunoglobulin (Ig) E and interleukin (IL)-2 were determined by radioimmunoassay. The activity of superoxide dismutase (SOD) in rat serum was determined by xanthine oxidation method. The activity of malondialdehyde (MDA) in serum of rats was determined by thiobarbituric acid (TBA) colorimetry. Result:Compared with blank group, intestinal mucosal injury score of rats in model group was significantly increased (PP3, FT4, IL-2 and SOD were significantly decreased (PPPPPP3, FT4, IL-2, and SOD contents were significantly increased (PPPPPConclusion:The unbalanced expressions of TLR4 and its negative regulating factor IRAK-M are connected with the pathogenesis of UC.Sishenwan can cure UC and control the expression of TLR4 and promote the expression of IRAK-M.
ABSTRACT
Objective:To investigate the expression of interleukin-1 receptor associated kinase-M(IRAKM)in systemic lupus erythematosus ( SLE) and its relationship with immunoregulation .Methods:103 patients with SLE were divided into active stage group (n=55) and stable stage group(n=48) according to their disease activity score (SLEDAI),and 40 healthy persons were chosen as control group.Real-time quantitative PCR ( RT-PCR ) was used to detect the expression of IRAKM mRNA in peripheral blood monouclear cells of the three groups .The levels of anti ds-DNA antibody and anti Sm antibody in serum were detected by Enzyme linked immunosorbent assay(ELISA).The levels of serum complement C3 and C4 were measured by immune scattering turbidimetry .Factor analysis of variance was performed to analyze the differences of all the observed indicators in the three groups .The correlation between IRAKM and autoantibodies and complements of SLE was analyzed by Pearson or Spearman .Results: The expression level of IRAKM mRNA of SLE patients in the active stage group and stable stage group were significantly lower than that in the control group ( P<0.05).Moreover,the expression level of IRAKM mRNA in the active stage group was lower than that in the stable stage group ,and the difference was statistically significant ( P<0.05 ) .Compared with the control group , the levels of anti ds-DNA antibody and anti Sm antibody in the active stage group and stable stage group were significantly increased ( P<0.05 ) ,and the levels of complement C 3 and C4 were obviously lower(P<0.05).The changes of serum levels of autoantibodies and complements in the active stage group were more remarkable than those in the stable stage group ( P<0.05 ) .The expression level of IRAKM mRNA in the SLE patients was negatively correlated with the levels of anti ds-DNA antibody and anti Sm antibody (P<0.05).However,it was positively correlated with the levels of complement C3 and C4 ( P<0.05 ) .Conclusion: IRAKM participates in the immune regulation process of SLE through negative regulation,and its expression level is closely related to the degree of SLE disease activity .
ABSTRACT
Objective To study the effect of Xuebijing, a Chinese traditional medicine injection, on the THP-1 cells challenged by endotoxin, and to explore whether it induces endotoxin tolerance. Methods The THP-1 cells were pretreated with Xuebijing in different concentrations (10, 25, 50 mg/mL) and times (4, 12, 24 hours), and then challenged by endotoxin. The level of TNF-α in culture supernatant was detected by ELISA assay, and the expression of TLR4 and IRAK-M mRNA were detected by Real time-PCR technique. Results There was no significant difference in TNF-α level among all the groups (pretreated with different concentrations of Xuebijing for different time) (P>0.05). Only in 50 mg/mL Xuebijing group, TLR4 mRNA was 1.547-fold increase in 24 h than in 4 h group (P<0.05). Only when pretreated for 24 h, IRAK-M mRNA was 1.349-fold increase in 50 mg/mL Xuebijing group than in control group (P<0.05). However,there was no significant difference among other groups (P>0.05). Conclusions Xuebijing can not block the release of TNF-α from the THP-1 cells challenged by endotoxin;and it does not induce endotoxin tolerance. When pretreated with high concentration of Xuebijing for long time, the expression of both TLR4 and IRAK-M mRNA is up-regulated, but its significance is not yet clear.