Huaiqihuang may protect from proteinuria by resisting MPC5 podocyte damage via targeting p-ERK/CHOP pathway

Authors

  • Tingxia Li The Children’s Hospital Zhejiang University School of Medicine, Hangzhou, Zhejiang, China
  • Jianhua Mao The Children’s Hospital Zhejiang University School of Medicine, Hangzhou, Zhejiang, China
  • Lei Huang The Children’s Hospital Zhejiang University School of Medicine, Hangzhou, Zhejiang, China
  • Haidong Fu The Children’s Hospital Zhejiang University School of Medicine, Hangzhou, Zhejiang, China
  • Shuohui Chen The Children’s Hospital Zhejiang University School of Medicine, Hangzhou, Zhejiang, China
  • Aimin Liu The Children’s Hospital Zhejiang University School of Medicine, Hangzhou, Zhejiang, China
  • Yuqin Liang The Children’s Hospital Zhejiang University School of Medicine, Hangzhou, Zhejiang, China

DOI:

https://doi.org/10.17305/bjbms.2016.887

Keywords:

Proteinuria, podocytes, Huaiqihuang, protective role, p-ERK/CHOP pathway

Abstract

The purpose of this study was to investigate the potential effects of Huaiqihuang (HQH) granule, a Chinese herbal medicine, in treating proteinuria and to reveal its possible mechanism. MPC5 podocytes were cultured in vitro at 37°C and induced with tunicamycin (TM). The TM-induced cells were treated with HQH at different concentrations. The cell proliferation was detected using the MTT assay. The optimal effective dose of HQH for MPC5 cells was determined by the MTT assay and LDH assay respectively. The influences of HQH on the proteinuria-related protein expression and the signaling pathway associated protein expression were also detected using quantitative reverse transcription PCR and Western blotting analysis. The results showed that the MPC5 cell model was successfully constructed in vitro. The HQH application could improve the harmful effects induced by TM on the MPC5 cells, including promoted cell proliferation and suppressed cell apoptosis. Furthermore, the protein expression, including podocin, nephrin, and synaptopodin was down-regulated by the TM treatment in the MPC5 cells. On contrary, the expression of these proteins was up-regulated after the HQH application. Also, the effect of TM on integrin α3 and integrin β1 expressions was also reversed by the HQH treatment. Moreover, the HQH application decreased the expression of p-ERK and DNA-damage-inducible transcript 3 (DDIT3 or CHOP) in the MPC5 cells, which was opposite to the effect observed in the cells treated with TM. Taken together, our study suggest that HQH application may protect podocytes from TM damage by suppressing the p-ERK/CHOP signaling pathway.

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References

Jafar TH, Stark PC, Schmid CH, Landa M, Maschio G, De Jong PE, et al. Progression of chronic kidney disease: the role of blood pressure control, proteinuria, and angiotensin-converting enzyme inhibition. Ann Intern Med. 2003;139(4):244-52. http://dx.doi.org/10.7326/0003-4819-139-4-200308190-00006.

Harita Y. Tyrosine kinase signaling in kidney glomerular podocytes. J Signal Transduct. 2011;2011:317852. http://dx.doi.org/10.1155/2011/317852.

Asanuma K, Yanagida-Asanuma E, Takagi M, Kodama F, Tomino Y. The role of podocytes in proteinuria. Nephrology 2007;12(Supplement s3):S15-20.

http://dx.doi.org/10.1111/j.1440-1797.2007.00876.x.

Harita Y. The Mechanisms of proteinuria: The role of signaling in podocytes. Nihon Shoni Jinzobyo Gakkai Zasshi 2007;20:176-81.

http://dx.doi.org/10.3165/jjpn.20.176.

Silva C, Inês LS, Nour D, Straub RH. Differential male and female adrenal cortical steroid hormone and cortisol responses to interleukin-6 in humans. Ann N Y Acad Sci. 2002;966(1):68-72. DOI: 10.1111/j.1749-6632.2002.tb04203.x.

Dong Y, Gao G, Jiao X, Wang X. HPLC-MS Determination of adrenal cortical hormone in traditional chinese medicine. Chinese Pharmaceutical Affairs. 2013.

Salama ZB. Pharmaceutical agent comprising blood components <10 Kda and their use for prophylaxis and treatment of defects of the immune system. United States patent US20080220083. 2008.

Carlotti AP, Franco PB, Elias LL, Facincani I, Costa EL, Foss N, et al. Glucocorticoid receptors, in vitro steroid sensitivity, and cytokine secretion in idiopathic nephrotic syndrome. Kidney Int. 2004;65(2):403-8.

DOI:10.1111/j.1523-1755.2004.00392.x.

Wen S, Zhi Z, Jian Y, Wang YH, Man X, Xia G, et al. Effects of Chinese herbal medicine Huaiqihuang Granule on nephrin and podocin expressions in renal tissues of rats with adriamycin-induced nephrosis. Chin J Integr Med. 2011;9(5):546-52. http://dx.doi.org/10.3736/jcim20110513.

Cao GH, Liu CH, Yun Y. Impacts of Huaiqihuang granules on humoral immunity and cellular immunity in children nephrotic syndrome. World Journal of Integrated Traditional & Western Medicine. 2013.

Wang X, Luan J, Tao LI, Wang X. Clinical study on Huaiqihuang granule on infantile nephropathy palindromia and its mechanism. Journal of Hubei University of Chinese Medicine. 2011.

Li LT, Shi MY, Wei SY, Li T, Li B. Huai Qi Huang ameliorates proteinuria and hematuria in mild IgA nephropathy patients: a prospective randomized controlled study. J Formos Med Assoc. 2013;112(12):766–72.

http://dx.doi.org/10.1016/j.jfma.2013.10.019.

Huber T. Molekulare pathogenese der proteinuria [In German]. Dtsch med Wochenschr 2008; 133(18): 954-58. http://dx.doi.org/10.1055/s-2008-1075673.

Ying MC, Miner JH. Glomerular basement membrane and related glomerular disease. Transl Res. 2012;160(4):291–7.

http://dx.doi.org/10.1016/j.trsl.2012.03.004.

Zhang B, Ning NI. Effect of Huaiqihuang particles on serum TNF-α, IL-18 and IL-10 in children with primary nephrotic syndrome. Chinese Journal of Practical Pediatrics. 2010;25(1).

Zhao CG, Yang YC, Wu YB, Zhang B. Changes of T regulatory cells and interleukin-10 in children with primary nephrotic syndrome before and after treatment assisted with Huaiqihuang particles. Journal of Applied Clinical Pediatrics. 2011;26(11):859-68.

Liu H, Sun W, Gu L, Tu Y, Yu B, Hu H. Huaiqihuang Granules reduce proteinuria by enhancing nephrin expression and regulating necrosis factor κB signaling pathway in adriamycin-induced nephropathy. Chin J Integr Med. 2015.

Arbabi S, Maier RV. Mitogen-activated protein kinases. Crit Care Med. 2002;30(1):S74-9. http://dx.doi.org/10.1097/00003246-200201001-00010.

Ma Y, Wei S, Peker D. An extremely rare primary gallbladder myxoid liposarcoma associated with amplification of DDIT3 gene. J Gastrointestin Liver Dis. 2014;23:460-1.

Miyazaki-Anzai S, Masuda M, Demos-Davies KM, Keenan AL, Saunders SJ, Masuda R, et al. Endoplasmic reticulum stress effector CCAAT/enhancer-binding protein homologous protein (CHOP) regulates chronic kidney disease-induced vascular calcification. J Am Heart Assoc. 2014;3(3):e000949.

http://dx.doi.org/10.1161/JAHA.114.000949.

Rodriguez-Agudo D, Calderon-Dominguez M, Medina MA, Ren S, Gil G, Pandak WM. ER stress increases StarD5 expression by stabilizing its mRNA and leads to relocalization of its protein from the nucleus to the membranes. J Lipid Res. 2012;141(12):586–7.

Reiko I, Masaomi N, Hiroshi O, Hiroshi U, Yasuko K, Kiyokazu N, et al. Involvement of endoplasmic reticulum (ER) stress in podocyte injury induced by excessive protein accumulation. Kidney Int. 2005;68(6):2639-50.

http://dx.doi.org/10.1111/j.1523-1755.2005.00736.x.

Du L, Shi P, Zhao ZJ 1,Wu Ge-dun-qi-qi-ge, Zhang XX, Lin R. Effect of Tangshenping on podocytes' proliferation and apoptosis in high glucose and LPS environment. China Journal of Traditional Chinese Medicine & Pharmacy 2013.

Radko L, Minta M, Stypuła-Trębas S. Cellular toxicity of malachite green and leucomalachite green evaluated on two rat cell lines by MTT, NRU, LDH, and protein assays. Bull Vet Inst Pulawy. 2011;55(2):347-53.

Han X, Gelein R, Corson N, Wade-Mercer P, Jiang J, Biswas P, et al. Validation of an LDH assay for assessing nanoparticle toxicity. Toxicology. 2011;287(1-3):99–104. http://dx.doi.org/10.1016/j.tox.2011.06.011.

Hummon AB, Lim SR, Difilippantonio MJ, Ried T. Isolation and solubilization of proteins after TRIzol® extraction of RNA and DNA from patient material following prolonged storage. Biotechniques. 2007;42(4):467. http://dx.doi.org/10.2144/000112401.

Mabuchi S, Altomare DA, Cheung M, Zhang L, Poulikakos PI, Hensley HH, et al. RAD001 inhibits human ovarian cancer cell proliferation, enhances cisplatin-induced apoptosis, and prolongs survival in an ovarian cancer model. Clin Cancer Res. 2007;13(14):4261-70. http://dx.doi.org/10.1158/1078-0432.CCR-06-2770.

Hermann PD, Wilhelm K, Matthias K. Cell biology of the glomerular podocyte. Physiol Rev. 2003;83(1):253-307. http://dx.doi.org/10.1152/physrev.00020.2002.

Samant RS, Clarke PA, Paul WE. E3 ubiquitin ligase Cullin-5 modulates multiple molecular and cellular responses to heat shock protein 90 inhibition in human cancer cells. Proc Natl Acad Sci U S A. 2014;111(18):6834-9. http://dx.doi.org/10.1073/pnas.1322412111.

Braun MH, Steele SL, Perry SF. The responses of zebrafish (Danio rerio) to high external ammonia and urea transporter inhibition: nitrogen excretion and expression of rhesus glycoproteins and urea transporter proteins. J Exp Biol. 2009;212(Pt23):3846-56. http://dx.doi.org/10.1242/jeb.034157.

Xu XM, Wang XB, Chen MM, Tao L, Li YX, Jia WH, et al. MicroRNA-19a and -19b regulate cervical carcinoma cell proliferation and invasion by targeting CUL5. Cancer Lett. 2012;322(2):148–58.

http://dx.doi.org/10.1016/j.canlet.2012.02.038.

Wu J, Zhang R, Torreggiani M, Ting A, Xiong H, Striker GE, et al. Induction of diabetes in aged C57B6 mice results in severe nephropathy: an association with oxidative stress, endoplasmic reticulum stress, and inflammation. Am J Pathol. 2010;176(5):2163-76. http://dx.doi.org/10.2353/ajpath.2010.090386.

Garovic VD, Wagner SJ, Petrovic LM, Gray CE, Pauline H, Hikaru S, et al. Glomerular expression of nephrin and synaptopodin, but not podocin, is decreased in kidney sections from women with preeclampsia. Nephrol Dial Transplant. 2007;22(4):1136-43. http://dx.doi.org/10.1093/ndt/gfl711.

Zhang H, Ding J, Fan Q, Liu S. RPC6 up-regulation in Ang II-induced podocyte apoptosis might result from ERK activation and NF-κB translocation. Exp Biol Med. 2009;234:1029-36. http://dx.doi.org/10.3181/0901-RM-11.

Fujita H, Omori S, Ishikura K, Hida M, Awazu M. ERK and p38 mediate high-glucose-induced hypertrophy and TGF-beta expression in renal tubular cells. Am J Physiol Renal Physiol. 2004;286(1):F120-6.

http://dx.doi.org/10.1152/ajprenal.00351.2002.

Huaiqihuang may protect from proteinuria by resisting MPC5 podocyte damage via targeting p-ERK/CHOP pathway

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Published

02-08-2016

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1.
Huaiqihuang may protect from proteinuria by resisting MPC5 podocyte damage via targeting p-ERK/CHOP pathway. Biomol Biomed [Internet]. 2016 Aug. 2 [cited 2024 Jul. 22];16(3):193-200. Available from: https://www.bjbms.org/ojs/index.php/bjbms/article/view/887