Inflammatory expression in IgA nephropathy cell model and anti-inflammatory effect of sodium valproate

doi:10.3969/j.issn.1674-8115.2022.06.009

Abstract

Abstract: Objective

·To study the effects of aggregated IgA1 (P-aIgA1) from patients with IgA nephropathy on the secretion of inflammatory factors and cell proliferation of human renal mesangial cells (HMCs), and observe the anti-inflammatory and anti-proliferation effects of histone deacetylation (HDAC) inhibitor sodium valproate (VPA) in vitro.

Methods

·N-IgA1 (IgA1 from health control) and P-IgA1 (IgA1 from patients with IgA nephropathy) were prepared by affinity chromatography, and P-aIgA1 and N-aIgA1 were prepared by thermal polymerization. The expression of inflammatory factors in HMCs culture supernatant was detected by human inflammatory factor protein chip. The proliferation of HMCs was detected by MTT assay. The expression of related proteins was detected by Western blotting or ELISA.

Results

·The results of inflammatory factor protein chip showed that the concentration of interleukin-6 soluble receptor (IL-6sR), regulated upon activation normal T cell expressed and secreted factor(RANTES), tissue inhibitor of metalloproteinase 1 (TIMP1), tissue inhibitor of metalloproteinase 2 (TIMP2), tumor necrosis factor-α(TNF-α) and tumor necrosis factor type Ⅱ receptor (TNFR Ⅱ) in the culture supernatant of HMCs in the P-aIgA1 group were significantly up-regulated compared with the control group, which were 34, 124, 269, 100, 1.6 and 52 times higher than those in the control group, respectively; N-aIgA1, P-IgA1 and P-aIgA1 could promote the expression of TIMP2, and there were significant differences (P=0.000, P=0.000, P=0.001). The proliferation of HMCs was observed by MTT method. The results showed that: ① Compared with the control group, N-IgA1 group had no significant effect on the proliferation of HMCs, and P-IgA1 and P-aIgA1 could significantly promote the proliferation of HMCs (P=0.045 and P=0.003); compared with the N-IgA1, HMCs in the P-aIgA1 group had significant proliferation, and the differences was statistically significant (P=0.036). ② The results of the effects of different concentrations of P-aIgA1 on the proliferation of HMCs showed that: 25 μg/mL P-aIgA1 could significantly promote the proliferation of HMCs (P=0.038), in a dose-dependent manner. ③ 400 μg/mL VPA could significantly inhibit the proliferation of HMCs (P=0.028). The effect of different IgA1 on HDAC1 in HMCs showed that the expression of HDAC1 in each group was (8.64±0.59) times (P-aIgA1), (5.42±0.16) times (P-IgA1) and (5.87±0.58) times (N-IgA1) higher than that in the control group, respectively; compared with N-IgA1, the expression of HDAC1 was significantly increased in the P-aIgA1 group (P=0.021). The effect of VPA on TNF-α secretion by mesangial cells showed that the protein expression of TNF-α in the P-aIgA1 group was significantly increased compared with the normal control group (P=0.001); compared with the P-aIgA1 group, the protein expression of TNF-α in the P-aIgA1+VPA group decreased significantly (P=0.035).

Conclusion

·P-aIgA1 can significantly promote the release of pro-inflammatory factors and mesangial cell proliferation, and the effect of promoting cell proliferation is concentration-dependent.Abnormal acetylation modification exists in IgA nephropathy cell model in vitro, which is involved in mesangial cell proliferation and inflammatory response. The HDAC inhibitor sodium valproate can partially reverse the above reaction. The above results suggest that sodium valproate has a certain application prospect in IgAN disease intervention, and provide a new idea for the prevention and treatment of IgA nephropathy from the perspective of epigenetics.

Key words: IgA nephropathy, human renal mesangial cell, histone deacetylation, sodium valproate, inflammatory factor

CLC Number:

R593.9

DAI Qin, WANG Weiming. Inflammatory expression in IgA nephropathy cell model and anti-inflammatory effect of sodium valproate[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022, 42(6): 751-757.

Figures/Tables 7

Fig 1 Expression of inflammatory factor in cultured HMCs supernatant induced by different IgA1s

Fig 2 Statistical chart of the array

Fig 3 Effects of different IgA1s on the proliferation of mesangial cell

Fig 4 Effects of different concentrations of P-aIgA1 on the proliferation of mesangial cell

Fig 5 Effects of different IgA1s on HDAC1 protein in HMCs

Fig 6 Effects of different concentrations of VPA on the proliferation of mesangial cell induced by P-aIgA1

Fig 7 Effects of different stimulators on TNF-α protein in HMCs cultured supernatant

TrendMD

References 22

1	LAI K N, HO R T, LEUNG J C, et al. Increased mRNA encoding for transforming factor-beta in CD4+ cells from patients with IgA nephropathy[J]. Kidney Int, 1994, 46(3): 862-868.
2	BERTHOUX F, SUZUKI H, THIBAUDIN L, et al. Autoantibodies targeting galactose-deficient IgA1 associate with progression of IgA nephropathy[J]. J Am Soc Nephrol, 2012, 23(9): 1579-1587.
3	戴芹, 缪怡, 王伟铭. 血浆Gd-IgA1浓度与IgA肾病严重程度的相关性及其产生机制研究[J]. 临床肾脏病杂志, 2020, 20(9): 711-716.
	DAI Q, MIAO Y, WANG W M. Study on the relationship between plasma Gd-IgA1 concentration and disease severity in IgA nephropathy and its production mechanism[J]. J Clin Nephrol, 2020,20(9):711-716.
4	GIANNAKAKIS K, FERIOZZI S, PEREZ M, et al. Aberrantly glycosylated IgA1 in glomerular immune deposits of IgA nephropathy[J]. J Am Soc Nephrol, 2007, 18(12): 3139-3146.
5	VAN DER BOOG P J M, VAN KOOTEN C, DE FIJTER J W, et al. Role of macromolecular IgA in IgA nephropathy[J]. Kidney Int, 2005, 67(3): 813-821.
6	XU L X, ZHAO M H. Aberrantly glycosylated serum IgA1 are closely associated with pathologic phenotypes of IgA nephropathy[J]. Kidney Int, 2005, 68(1): 167-172.
7	DING J X, XU L X, LV J C, et al. Aberrant sialylation of serum IgA1 was associated with prognosis of patients with IgA nephropathy[J]. Clin Immunol, 2007, 125(3): 268-274.
8	TOMANA M, NOVAK J, JULIAN B A, et al. Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and antiglycan antibodies[J]. J Clin Invest, 1999, 104(1): 73-81.
9	ABBOUD H E. Mesangial cell biology[J]. Exp Cell Res, 2012, 318(9): 979-985.
10	MOURA I C, BENHAMOU M, LAUNAY P, et al. The glomerular response to IgA deposition in IgA nephropathy[J]. Semin Nephrol, 2008, 28(1): 88-95.
11	INOUE M K, YAMAMOTOYA T, NAKATSU Y, et al. The xanthine oxidase inhibitor febuxostat suppresses the progression of IgA nephropathy, possibly via its anti-inflammatory and anti-fibrotic effects in the gddY mouse model[J]. Int J Mol Sci, 2018, 19(12): 3967.
12	DAI Q, LIU J, DU Y L, et al. Histone deacetylase inhibitors attenuate P-aIgA1-induced cell proliferation and extracellular matrix synthesis in human renal mesangial cells in vitro[J]. Acta Pharmacol Sin, 2016, 37(2): 228-234.
13	SUZUKI H, OHTO U, HIGAKI K, et al. Structural basis of pharmacological chaperoning for human β-galactosidase[J]. J Biol Chem, 2014, 289(21): 14560-14568.
14	CHEN S W, BELLEW C, YAO X, et al. Histone deacetylase (HDAC) activity is critical for embryonic kidney gene expression, growth, and differentiation[J]. J Biol Chem, 2011, 286(37): 32775-32789.
15	SELVASKANDAN H, SHI S F, TWAIJ S, et al. Monitoring immune responses in IgA nephropathy: biomarkers to guide management[J]. Front Immunol, 2020, 11: 572754.
16	BUSH E W, MCKINSEY T A. Protein acetylation in the cardiorenal axis: the promise of histone deacetylase inhibitors[J]. Circ Res, 2010, 106(2): 272-284.
17	HADDEN M J, ADVANI A. Histone deacetylase inhibitors and diabetic kidney disease[J]. Int J Mol Sci, 2018, 19(9): 2630.
18	戴芹, 王伟铭. 组蛋白乙酰化在IgA肾病发病中的作用[J]. 上海交通大学学报(医学版), 2020, 40(8): 1063-1068.
	DAI Q, WANG W M. Role of histone acetylation in the pathogenesis of IgA nephropathy[J]. J Shanghai Jiao Tong Univ (Med Sci), 2020,40(8): 1063-1068.
19	JONES J, JUENGEL E, MICKUCKYTE A, et al. The histone deacetylase inhibitor valproic acid alters growth properties of renal cell carcinoma in vitro and in vivo[J]. J Cell Mol Med, 2009, 13(8B): 2376-2385.
20	SANAEI M, KAVOOSI F. Effect of valproic acid on the class I histone deacetylase 1, 2 and 3, tumor suppressor genes p21WAF1/CIP₁ and p53, and intrinsic mitochondrial apoptotic pathway, pro- (bax, bak, and bim) and anti- (bcl-2, bcl-xL, and mcl-1) apoptotic genes expression, cell viability, and apoptosis induction in hepatocellular carcinoma HepG2 cell line[J]. Asian Pac J Cancer Prev, 2021, 22(S1): 89-95.
21	RIVA G, CILIBRASI C, BAZZONI R, et al. Valproic acid inhibits proliferation and reduces invasiveness in glioma stem cells through Wnt/β catenin signalling activation[J]. Genes, 2018, 9(11): 522.
22	VAN BENEDEN K, GEERS C, PAUWELS M, et al. Valproic acid attenuates proteinuria and kidney injury[J]. J Am Soc Nephrol, 2011, 22(10): 1863-1875.

[1]	ZHANG Huanyu, JIANG Yiting, ZHU Xiaochen, HE Zhiyan, ZHOU Wei, SONG Zhongchen. Effects of gingipain extracts on brain neuroinflammation in mice [J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022, 42(5): 570-577.
[2]	DAI Qin1, WANG Wei-ming2. Role of histone acetylation in the pathogenesis of IgA nephropathy [J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2020, 40(8): 1063-1068.
[3]	KONG Ling-yun, JIANG Geng-ru. Effect of serum IgA1 on human mesangial cells and of TGF-β1 in patients of IgA nephropathy with different renal function progress [J]. , 2018, 38(6): 647-.
[4]	SU Zheng-jia, PAN Xiao-qian, CAO Jiu-mei, WU Fang. Effects of liraglutide on glucose induced of miRNA-146b-3p, inflammatory factors and COX-2 in human umbilical vein endothelial cells [J]. , 2018, 38(12): 1420-.
[5]	JING Bo, CHEN Peng-hui, GAO Xiang, XU Yuan-yuan, WU Yun-zhao, SUN Yun, WU Ying-li . Establishment of cell-based screening system for compound regulating the stability of retinoic acid receptors [J]. , 2017, 37(4): 432-.
[6]	SHI Ying-feng, XU Liu-qing, ZHUANG Shou-gang, LIU Na . Correlation between pathology and clinical indexes in IgA nephropathy patients#br# [J]. , 2017, 37(10): 1362-.
[7]	YANG Meng, XIE Jing-yuan, OUYANG Yan, ZHANG Xiao-yan, PAN Xiao-xia, XU Jing, WANG Zhao-hui, WANG Wei-ming, CHEN Nan. Analysis of clinicopathologic features and prognosis of IgA nephropathy patients with H factor deposition in renal tissue [J]. , 2016, 36(8): 1144-.
[8]	ZHOU Yue-ling, JIANG Geng-ru. Research progresses of clinical prediction of development of IgA nephropathy towards end-stage renal disease [J]. , 2016, 36(2): 296-.
[9]	LIAO Wei-wen, SONG Zhong-chen, SHU Rong, et al. Effects of recombinant human amelogenin on expressions of inflammatory factors of human periodontal ligament fibroblasts under inflammatory microenvironment [J]. , 2016, 36(1): 33-.
[10]	GU Yu, LIU Shuang, ZHU Yi, et al. Establishment and identification of a mice model of transgenic IgA nephropathy [J]. , 2016, 36(1): 43-.
[11]	HUANG Wei, WANG Wei-ming. Applications and research progresses of new immunosuppressive agents for the treatment of IgA nephropathy [J]. , 2015, 35(11): 1722-.
[12]	DAI Qin, LIU Jian, HAO Xu, et al. Protective effect of sodium valproate on rats with adriamycin-induced nephropathy [J]. , 2014, 34(5): 573-.
[13]	ZHANG Ying, ZHONG Wen-hui, WANG Ai-zhong. Effects of dexmedetomidine on function of intestinal mucosal barrier of rats with ischemia-reperfusion injury [J]. , 2014, 34(4): 487-.
[14]	WANG Wei-ming, JIA Xiao-yuan, PAN Xiao-xia, et al. Clinical study on treatment of IgA nephropathy with renal insufficiency by corticosteroid, corticosteroid combined with cyclophosphamide and corticosteroid combined with mycophenolate mofetil [J]. , 2013, 33(2): 162-.
[15]	WANG Xi-jin, YAN Zhi-qiang, GU Sheng-li, et al. Minocycline protects dopaminergic neurons by inhibiting production of proinflammatory factors [J]. , 2012, 32(6): 711-.