TLR9对IgA肾病肾小管上皮细胞炎症反应及Gd-IgA1水平的影响实验研究OA

Objective:To investigate the effects of Toll-like receptor 9(TLR9)on inflammatory response and galactose-deficient IgA1(Gd-IgA1)levels in renal tubular epithelial cells of immunoglobulin A nephropathy(IgAN).Methods:Human renal tubular epithelial cells HK-2 were selected and divided into six groups:control group(routine culture),model group(treated with 25 μg/ml IgA),TLR9 activation group(treated with 25 μg/ml IgA+5 μg/ml TLR9 agonist CpG oligodeoxynucleotide[CpG-ODN]),TLR9 silencing group(transfected with TLR9 siRNA+25 μg/ml IgA),NC group(treated with 25 μg/ml IgA+5 μg/ml Non-CpG-ODN),and inhibition NC group(trans-fected with negative control siRNA+25 μg/ml IgA).Cell proliferation ability of HK-2 cells in each group was detec-ted by CCK-8 assay.The mRNA expressions of a proliferation-inducing ligand(APRIL),B cell activating factor(BAFF),and TLR9 in each group were detected by RT-qPCR.The levels of APRIL,BAFF and Gd-IgA1 in each group were detected by ELISA.The protein expressions of APRIL,BAFF,TLR9 and Gd-IgA1 in each group were detected by Western blot.Results:Comparison of HK-2 cell proliferation rates at 24 and 48 hours among groups showed that the TLR9 activation group was lower than the model group,and the model group was lower than the control group;the TLR9 silencing group was higher than both the model group and TLR9 activation group,but lower than the control group(all P＜0.05).Comparisons of TLR9,BAFF and APRIL mRNA and protein expression levels,BAFF,APRIL and Gd-IgA1 contents,and Gd-IgA1 protein expression levels in HK-2 cells among groups showed that the TLR9 activation group was higher than the model group,the model group was higher than the control group,and the control group was higher than the TLR9 silencing group(all P＜0.05).Conclusion:TLR9 may participate in the occurrence and development of IgAN by inhibiting the proliferation of renal tubular epithelial cells,enhancing inflammatory re-sponse,and promoting the production of pathogenic Gd-IgA1.