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Research Article|Articles in Press, 155376

Inhibition of S100A8/A9 ameliorates renal interstitial fibrosis in diabetic nephropathy

Published:December 11, 2022DOI:https://doi.org/10.1016/j.metabol.2022.155376

      Highlights

      • S100A8/A9 expressions are increased in diabetic renal tubular.
      • The levels of S100A8/A9 are associated with renal interstitial fibrosis.
      • S100A8/A9 promote renal interstitial fibrosis through TLR4/NF-κB signal pathway.
      • New compound AB38b ameliorates renal interstitial fibrosis by inhibiting S100A8/A9.

      Abstract

      Background

      Renal interstitial fibrosis (RIF) is one of the main features of diabetic nephropathy (DN), but the molecular mechanisms mediating RIF in DN has yet been fully understood. S100A8 and S100A9 are the proteins associated with immune and inflammation response. Here we reported the expression of S100A8 and S100A9 were significantly increased on tubular epithelial cells in diabetic kidneys through a proteomic analysis.

      Methods

      We detected the expression of S100A8/A9 in diabetic kidneys by using immunoblotting, real-time PCR and immunostaining. RNA silencing and overexpression were performed by using S100A8/A9 expression/knockdown lentivirus to investigate the connection between S100A8/A9 and epithelial to mesenchymal transition (EMT) process. We also identify the expression of TLR4/NFκB pathway-related molecules in the case mentioned above. Afterwards a CO-IP assay was used to verify that compound AB38b ameliorates the EMT by interfering S100A8/A9 expression.

      Results

      The expression of S100A8 and S100A9 were significantly increased on tubular epithelial cells in diabetic kidneys. S100A8/A9 knocking-down alleviate and over-expression promote the renal interstitial fibrosis of diabetic mice. Mechanically, high levels of S100A8/A9 expression in tubular epithelial cells during diabetic condition activated the TLR4/NF-κB signal pathway which promoted the EMT process and finally led to RIF progression. S100A8/A9 knockdown ameliorated RIF of diabetic mice. Further experiments revealed that compound AB38b inhibited the EMT progression of tubular epithelial cells induced by S100A8/A9 through interfering the expressions of S100A8/A9.

      Conclusions

      Our study suggest that abnormal expression of S100A8/A9 in the disease condition promotes EMT process and RIF through TLR4/NF-κB signal pathway. Using small molecular inhibitor AB38b to inhibit the abnormal expressions of S100A8/A9 might be a novel therapeutic strategy in treating DN.

      Graphical abstract

      Keywords

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      References

        • Umanath K.
        • Lewis J.B.
        Update on diabetic nephropathy: core curriculum 2018.
        Am J Kidney Dis. 2018; 71: 884-895
        • Gnudi L.
        The patient with diabetes mellitus.
        in: Turner N. Oxford textbook of clinical nephrology. Oxford University Press, 2016: 1199-1247
        • Chan S.C.
        • Zhang Y.
        • Shao A.
        • Avdulov S.
        • Herrera J.
        • Aboudehen K.
        • et al.
        Mechanism of fibrosis in HNF1B-related autosomal dominant tubulointerstitial kidney disease.
        J Am Soc Nephrol. 2018; 29: 2493-2509
        • Hills C.E.
        • Siamantouras E.
        • Smith S.W.
        • Cockwell P.
        • Liu K.K.
        • Squires P.E.
        TGFβ modulates cell-to-cell communication in early epithelial-to-mesenchymal transition.
        Diabetologia. 2012; 55: 812-824
        • Lu Q.
        • Ji X.J.
        • Zhou Y.X.
        • Yao X.Q.
        • Liu Y.Q.
        • Zhang F.
        • et al.
        Quercetin inhibits the mTORC1/p70S6K signaling-mediated renal tubular epithelial-mesenchymal transition and renal fibrosis in diabetic nephropathy.
        Pharmacol Res. 2015; 99: 237-247
        • Kanwar Y.S.
        • Sun L.
        • Xie P.
        • Liu F.Y.
        • Chen S.
        A glimpse of various pathogenetic mechanisms of diabetic nephropathy.
        Annu Rev Pathol. 2011; 6: 395-423
        • Zhao Y.
        • Yin Z.
        • Li H.
        • Fan J.
        • Yang S.
        • Chen C.
        • et al.
        MiR-30c protects diabetic nephropathy by suppressing epithelial-to-mesenchymal transition in db/db mice.
        Aging Cell. 2017; 16: 387-400
        • Yang T.
        • Shu F.
        • Yang H.
        • Heng C.
        • Zhou Y.
        • Chen Y.
        • et al.
        YY1: a novel therapeutic target for diabetic nephropathy orchestrated renal fibrosis.
        Metabolism. 2019; 96: 33-45
        • Sreejit G.
        • Abdel-Latif A.
        • Athmanathan B.
        • Annabathula R.
        • Dhyani A.
        • Noothi S.K.
        • et al.
        Neutrophil-derived S100A8/A9 amplify granulopoiesis after myocardial infarction.
        Circulation. 2020; 141: 1080-1094
        • Pruenster M.
        • Vogl T.
        • Roth J.
        S100A8/A9: from basic science to clinical application.
        Pharmacol Ther. 2016; 167: 120-131
        • Pepper R.J.
        • Wang H.H.
        • Rajakaruna G.K.
        • Papakrivopoulou E.
        • Vogl T.
        • Pusey C.D.
        • et al.
        S100A8/A9 (calprotectin) is critical for development of glomerulonephritis and promotes inflammatory leukocyte-renal cell interactions.
        Am J Pathol. 2015; 185: 1264-1274
        • Vogl T.
        • Gharibyan A.L.
        Pro-inflammatory S100A8 and S100A9 proteins: self-assembly into multifunctional native and amyloid complexes.
        Int J Mol Sci. 2012; 13: 2893-2917
        • Donato R.
        • Cannon B.R.
        • Sorci G.
        • Riuzzi F.
        • Hsu K.
        • Weber D.J.
        • et al.
        Functions of S100 proteins.
        Curr Mol Med. 2013; 13: 24-57
        • Lovisa S.
        • Zeisberg M.
        • Kalluri R.
        Partial epithelial-to-mesenchymal transition and other new mechanisms of kidney fibrosis.
        Trends Endocrinol Metab. 2016; 27: 681-695
        • Tammaro A.
        • Florquin S.
        • Brok M.
        • Claessen N.
        • Butter L.M.
        • Teske G.J.D.
        • et al.
        S100A8/A9 promotes parenchymal damage and renal fibrosis in obstructive nephropathy.
        Clin Exp Immunol. 2018; 193: 361-375
        • Zhong A.
        • Xu W.
        • Zhao J.
        • Xie P.
        • Jia S.
        • Sun J.
        • et al.
        S100A8 and S100A9 are induced by decreased hydration in the epidermis and promote fibroblast activation and fibrosis in the dermis.
        Am J Pathol. 2016; 186: 109-122
        • Ehlermann P.
        • Eggers K.
        • Bierhaus A.
        • Most P.
        • Weichenhan D.
        • Greten J.
        • et al.
        Increased proinflammatory endothelial response to S100A8/A9 after preactivation through advanced glycation end products.
        Cardiovasc Diabetol. 2006; 5: 6
        • Vogl T.
        • Tenbrock K.
        • Ludwig S.
        • Leukert N.
        • Ehrhardt C.
        • van Zoelen M.A.
        • et al.
        Mrp8 and Mrp14 are endogenous activators of Toll-like receptor 4, promoting lethal, endotoxin-induced shock.
        Nat Med. 2007; 13: 1042-1049
        • Bierhaus A.
        • Schiekofer S.
        • Schwaninger M.
        • Andrassy M.
        • Humpert P.M.
        • Chen J.
        • et al.
        Diabetes-associated sustained activation of the transcription factor nuclear factor-kappaB.
        Diabetes. 2001; 50: 2792-2808
        • Liu P.
        • Li F.
        • Qiu M.
        • He L.
        Expression and cellular distribution of TLR4, MyD88, and NF-κB in diabetic renal tubulointerstitial fibrosis, in vitro and in vivo.
        Diabetes Res Clin Pract. 2014; 105: 206-216
        • Liu B.
        • Ding F.
        • Hu D.
        • Zhou Y.
        • Long C.
        • Shen L.
        • et al.
        Human umbilical cord mesenchymal stem cell conditioned medium attenuates renal fibrosis by reducing inflammation and epithelial-to-mesenchymal transition via the TLR4/NF-κB signaling pathway in vivo and in vitro.
        Stem Cell Res Ther. 2018; 9: 7
        • Ma Z.J.
        • Zhang X.N.
        • Li L.
        • Yang W.
        • Wang S.S.
        • Guo X.
        • et al.
        Tripterygium glycosides tablet ameliorates renal tubulointerstitial fibrosis via the toll-like receptor 4/nuclear factor kappa B signaling pathway in high-fat diet fed and streptozotocin-induced diabetic rats.
        J Diabetes Res. 2015; 2015390428
        • Lin M.
        • Yiu W.H.
        • Li R.X.
        • Wu H.J.
        • Wong D.W.
        • Chan L.Y.
        • et al.
        The TLR4 antagonist CRX-526 protects against advanced diabetic nephropathy.
        Kidney Int. 2013; 83: 887-900
        • Hills C.E.
        • Siamantouras E.
        • Smith S.W.
        • Cockwell P.
        • Liu K.K.
        • Squires P.E.
        TGFβ modulates cell-to-cell communication in early epithelial-to-mesenchymal transition.
        Diabetologia. 2012; 55: 812-824
        • Du L.
        • Wang L.
        • Wang B.
        • Wang J.
        • Hao M.
        • Chen Y.B.
        • et al.
        A novel compound AB38b attenuates oxidative stress and ECM protein accumulation in kidneys of diabetic mice through modulation of Keap1/Nrf2 signaling.
        Acta Pharmacol Sin. 2020; 41: 358-372
        • Du L.
        • Wang J.
        • Chen Y.B.
        • Li X.Z.
        • Wang L.
        • Li Y.
        • et al.
        Novel biphenyl diester derivative AB-38b inhibits NLRP3 inflammasome through Nrf2 activation in diabetic nephropathy.
        Cell Biol Toxicol. 2020; 36: 243-260
        • Simonson M.S.
        Phenotypic transitions and fibrosis in diabetic nephropathy.
        Kidney Int. 2007; 71: 846-854
        • Yamamoto Y.
        • Kato I.
        • Doi T.
        • Yonekura H.
        • Ohashi S.
        • Takeuchi M.
        • et al.
        Development and prevention of advanced diabetic nephropathy in RAGE-overexpressing mice.
        J Clin Invest. 2001; 108: 261-8
        • Lin M.
        • Yiu W.H.
        • Wu H.J.
        • Chan L.Y.
        • Leung J.C.
        • Au W.S.
        • et al.
        Toll-like receptor 4 promotes tubular inflammation in diabetic nephropathy.
        J Am Soc Nephrol. 2012; 23: 86-102
        • Chen Y.
        • Sumardika I.W.
        • Tomonobu N.
        • Kinoshita R.
        • Inoue Y.
        • Iioka H.
        • et al.
        Critical role of the MCAM-ETV4 axis triggered by extracellular S100A8/A9 in breast cancer aggressiveness.
        Neoplasia. 2019; 21: 627-640
        • Dessing M.C.
        • Tammaro A.
        • Pulskens W.P.
        • Teske G.J.
        • Butter L.M.
        • Claessen N.
        • et al.
        The calcium-binding protein complex S100A8/A9 has a crucial role in controlling macrophage-mediated renal repair following ischemia/reperfusion.
        Kidney Int. 2015; 87: 85-94
        • Fujiu K.
        • Manabe I.
        • Nagai R.
        Renal collecting duct epithelial cells regulate inflammation in tubulointerstitial damage in mice.
        J Clin Invest. 2011; 121: 3425-3441
        • Kuwabara T.
        • Mori K.
        • Mukoyama M.
        • et al.
        Exacerbation of diabetic nephropathy by hyperlipidaemia is mediated by toll-like receptor 4 in mice.
        Diabetologia. 2012; 55: 2256-2266
        • Kuwabara T.
        • Mori K.
        • Kasahara M.
        • et al.
        Predictive significance of kidney myeloid-related protein 8 expression in patients with obesity- or type 2 diabetes-associated kidney diseases.
        PLoS One. 2014; 9e88942
        • Wu Y.
        • Li Y.
        • Zhang C.
        • A X.
        • Wang Y.
        • Cui W.
        • et al.
        S100a8/a9 released by CD11b+Gr1+ neutrophils activates cardiac fibroblasts to initiate angiotensin II-Induced cardiac inflammation and injury.
        Hypertension. 2014; 63: 1241-1250
        • Li R.
        • Guo Y.
        • Zhang Y.
        • Zhang X.
        • Zhu L.
        • Yan T.
        Salidroside ameliorates renal interstitial fibrosis by inhibiting the TLR4/NF-κB and MAPK signaling pathways.
        Int J Mol Sci. 2019; 20: 1103
        • Oldfield M.D.
        • Bach L.A.
        • Forbes J.M.
        • Nikolic-Paterson D.
        • McRobert A.
        • Thallas V.
        • et al.
        Advanced glycation end products cause epithelial-myofibroblast transdifferentiation via the receptor for advanced glycation end products (RAGE).
        J Clin Invest. 2001; 108: 1853-1863
        • Pinzi L.
        • Rastelli G.
        Molecular docking: shifting paradigms in drug discovery.
        Int J Mol Sci. 2019; 20: 4331
        • Dong L.
        • Zhang X.
        • Xiang W.
        • Ni J.
        • Zhou W.
        • Li H.
        Post-transcription mediated snail stabilization is involved in radiation exposure induced invasion and migration of hepatocarcinoma cells.
        Biomed Pharmacother. 2018; 103: 767-772
        • Li Y.
        • Chen B.
        • Yang X.
        • Zhang C.
        • Jiao Y.
        • Li P.
        • et al.
        S100a8/a9 signaling causes mitochondrial dysfunction and cardiomyocyte death in response to ischemic/reperfusion injury.
        Circulation. 2019; 140: 751-764
        • Al Fayi M.
        • Otifi H.
        • Alshyarba M.
        • Dera A.A.
        • Rajagopalan P.
        Thymoquinone and curcumin combination protects cisplatin-induced kidney injury, nephrotoxicity by attenuating NFκB, KIM-1 and ameliorating Nrf2/HO-1 signalling.
        J Drug Target. 2020; 5: 1-10
        • Ghosh S.
        • Choudhury S.
        • Chowdhury O.
        • et al.
        Inflammation-induced behavioral changes is driven by alterations in Nrf2-dependent apoptosis and autophagy in mouse hippocampus: role of fluoxetine.
        Cell Signal. 2020; 68109521
        • Chen Y.J.
        • Tang Z.Z.
        • Du L.
        • Liu Y.
        • Lu Q.
        • Ma T.F.
        • et al.
        A novel compound AB-38b improves diabetes-associated cognitive decline in mice via activation of Nrf2/ARE pathway.
        Brain Res Bull. 2019; 150: 160-167
        • Tan X.
        • Zheng X.
        • Huang Z.
        • Lin J.
        • Xie C.
        • Lin Y.
        Involvement of S100A8/A9-TLR4-NLRP3 inflammasome pathway in contrast-induced acute kidney injury.
        Cell Physiol Biochem. 2017; 43: 209-222