The role of C1GALT1C1 in lipopolysaccharide-induced IgA1 aberrant O-glycosylation in IgA nephropathy.
The role of C1GALT1C1 in lipopolysaccharide-induced IgA1 aberrant O-glycosylation in IgA nephropathy.
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DOI：
10.25011/cim.v33i1.11832
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发表时间：
2010-02
期刊：
影响因子：
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通讯作者：
Linshen Xie;W. Qin;Junming Fan;Jun Huang;Xi-sheng Xie;Zi Li
中科院分区：
文献类型：
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作者：
Linshen Xie;W. Qin;Junming Fan;Jun Huang;Xi-sheng Xie;Zi Li
目的IgA 1异常O-糖基化是伊加肾病（IgAN）的主要发病特征之一。本研究试图确定C1 GALT 1C 1在脂多糖（LPS）诱导的异常IgA 1 O-糖基化中的作用，并确定IgAN的潜在治疗靶点。方法22例IgAN患者和17例正常人的外周血淋巴细胞分别加入LPS和5-AZA培养3 ~ 7 d。采用实时荧光定量PCR和Western blot方法分别检测C1 GALT 1C 1 mRNA和蛋白的表达水平。ELISA和Vicia villosa（VV）凝集素结合试验测定IgA 1浓度和O-糖基化水平。对C1 GALT 1C 1蛋白表达与IgA 1 O-糖基化进行相关性分析。结果LPS刺激后IgAN患者淋巴细胞分泌的IgA 1明显高于正常对照组（P分别为0.26、0.002和0.005），而5-AZA可抑制IgA 1的分泌（P分别为0.001、0.025和0.001）。LPS刺激可明显抑制IgAN患者C1 GALT 1C 1的表达（第3、5和7天分别降低71%、82%和92%; P0.05）。结论LPS可诱导IgA肾病患者IgA 1异常O-糖基化，并抑制C1 GALT 1C 1的表达。5-AZA上调C1 GALT 1C 1表达可逆转IgA 1异常O-糖基化。这些结果表明，C1 GALT 1C 1可能在调节IgA 1 O-糖基化中起关键作用。
PURPOSE IgA1 aberrant O-glycosylation is one of the main pathogenetic features of IgA nephropathy (IgAN). This study attempted to determine the role of C1GALT1C1 in aberrant IgA1 O-glycosylation induced by lipopolysaccharide (LPS) and identify potential therapeutic targets in IgAN. METHODS Lymphocytes isolated from 22 patients with IgAN and 17 normal controls were cultured for 3 to 7 days with or without LPS and 5-azacytidine (5-AZA). Expression levels of C1GALT1C1 mRNA and protein were measured by real-time PCR and Western blot analysis, respectively. Concentration of IgA1 and level of O-glycosylation were determined by ELISA and Vicia villosa (VV) lectin-binding assay. Correlation analysis was performed between the expression of C1GALT1C1 protein and IgA1 O-glycosylation. RESULTS Lymphocytes from patients with IgAN secreted more IgA1 than that from normal controls after LPS stimulation (P=0.26, 0.002 and 0.005 on the 3rd, 5th and 7th day, respectively) which could be inhibited by 5-AZA (P=0.001, 0.025 and 0.001 on the 3rd, 5th and 7th day, respectively). Moreover, LPS stimulation could obviously inhibit C1GALT1C1 expression in patients with IgAN (decreased by 71%, 82% and 92% on the 3rd, 5th and 7th day, respectively; P 0.05). CONCLUSION LPS induced IgA1 aberrant O-glycosylation and suppressed C1GALT1C1 expression in patients with IgAN. Upregulation of C1GALT1C1 expression by 5-AZA could reverse the IgA1 aberrant O-glycosylation. These results suggest that C1GALT1C1 may play a key role in the regulation of IgA1 O-glycosylation.