Notoginsenoside R1 attenuates glucose-induced podocyte injury via the inhibition of apoptosis and the activation of autophagy through the PI3K/Akt/mTOR signaling pathway

Huang,Guodong; Zou,Bingyu; Lv,Jianzhen; Li,Tongyu; Huai,Guoli; Xiang,Shaowei; Lu,Shilong; Luo,Huan; Zhang,Yaping; Jin,Yi; Wang,Yi

doi:10.3892/ijmm.2017.2864

Notoginsenoside R1 attenuates glucose-induced podocyte injury via the inhibition of apoptosis and the activation of autophagy through the PI3K/Akt/mTOR signaling pathway

Authors:
- Guodong Huang
- Bingyu Zou
- Jianzhen Lv
- Tongyu Li
- Guoli Huai
- Shaowei Xiang
- Shilong Lu
- Huan Luo
- Yaping Zhang
- Yi Jin
- Yi Wang
View Affiliations

Affiliations: Department of Nephrology, Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning, Guangxi 530011, P.R. China, Department of Gynecology, Sichuan Academy of Medical Science and Sichuan Provincial People's Hospital, Chengdu, Sichuan 610072, P.R. China, School of Basic Medical Science, Guangxi University of Chinese Medicine, Nanning, Guangxi 530001, P.R. China, Department of Biomedical Engineering, Medical School of University of Electronic Science and Technology of China, Chengdu, Sichuan 610054, P.R. China, Center for Perinatal Research, Research Institute at Nationwide Children's Hospital, Columbus, OH 43215, USA
Published online on: January 20, 2017 https://doi.org/10.3892/ijmm.2017.2864
Pages: 559-568
Copyright: © Huang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Injury to terminally differentiated podocytes contributes ignificantly to proteinuria and glomerulosclerosis. The aim of this study was to examine the protective effects of notoginsenoside R1 (NR1) on the maintenance of podocyte number and foot process architecture via the inhibition of apoptosis, the induction of autophagy and the maintenance pf podocyte biology in target cells. The effects of NR1 on conditionally immortalized human podocytes under high glucose conditions were evaluated by determining the percentage apoptosis, the percentage autophagy and the expression levels of slit diaphragm proteins. Our results revealed that NR1 protected the podocytes against high glucose-induced injury by decreasing apoptosis, increasing autophagy and by promoting cytoskeletal recovery. The phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling pathway was further investigated in order to elucidate the mechanisms responsible for the protective effects of NR1 on podocytes. Our data indicated that treatment with NR increased the phosphorylation levels of PI3K, Akt and mTOR, leading to the activation of the PI3K/Akt/mTOR signaling pathway in podocytes. To the best of our knowledge, this is the first in vitro study to demonstrate that NR1 protects podocytes by activating the PI3K/Akt/mTOR pathway.

View Figures

View References

1	Mogensen CE: Microalbuminuria as a predictor of clinical diabetic nephropathy. Kidney Int. 31:673–689. 1987. View Article : Google Scholar : PubMed/NCBI
2	Rudberg S, Persson B and Dahlquist G: Increased glomerular filtration rate as a predictor of diabetic nephropathy - an 8-year prospective study. Kidney Int. 41:822–828. 1992. View Article : Google Scholar : PubMed/NCBI
3	White KE, Bilous RW and Diabiopsies Study Group: Structural alterations to the podocyte are related to proteinuria in type 2 diabetic patients. Nephrol Dial Transplant. 19:1437–1440. 2004. View Article : Google Scholar : PubMed/NCBI
4	Kriz W, Gretz N and Lemley KV: Progression of glomerular diseases: is the podocyte the culprit. Kidney Int. 54:687–697. 1998. View Article : Google Scholar : PubMed/NCBI
5	Dalla Vestra M, Masiero A, Roiter AM, Saller A, Crepaldi G and Fioretto P: Is podocyte injury relevant in diabetic nephropathy? Studies in patients with type 2 diabetes. Diabetes. 52:1031–1035. 2003. View Article : Google Scholar : PubMed/NCBI
6	Meyer TW, Bennett PH and Nelson RG: Podocyte number predicts long-term urinary albumin excretion in Pima Indians with Type II diabetes and microalbuminuria. Diabetologia. 42:1341–1344. 1999. View Article : Google Scholar : PubMed/NCBI
7	Pavenstadt H, Kriz W and Kretzler M: Cell biology of the glomerular podocyte. Physiol Rev. 83:253–307. 2003. View Article : Google Scholar
8	Huang G, Lv J, Li T, Huai G, Li X, Xiang S, Wang L, Qin Z, Pang J, Zou B and Wang Y: Notoginsenoside R1 ameliorates podocyte injury in rats with diabetic nephropathy by activating the PI3K/Akt signaling pathway. Int J Mol Med. 38:1179–1189. 2016.PubMed/NCBI
9	Quaggin SE: Transcriptional regulation of podocyte specification and differentiation. Microsc Res Tech. 57:208–211. 2002. View Article : Google Scholar : PubMed/NCBI
10	White KE, Bilous RW, Marshall SM, El Nahas M, Remuzzi G, Piras G, De Cosmo S and Viberti G: Podocyte number in normotensive type 1 diabetic patients with albuminuria. Diabetes. 51:3083–3089. 2002. View Article : Google Scholar : PubMed/NCBI
11	Verzola D, Gandolfo MT, Ferrario F, Rastaldi MP, Villaggio B, Gianiorio F, Giannoni M, Rimoldi L, Lauria F, Miji M, et al: Apoptosis in the kidneys of patients with type II diabetic nephropathy. Kidney Int. 72:1262–1272. 2007. View Article : Google Scholar : PubMed/NCBI
12	Liu WJ, Tang HT, Jia YT, Ma B, Fu JF, Wang Y, Lv KY and Xia ZF: Notoginsenoside R1 attenuates renal ischemia-reperfusion injury in rats. Shock. 34:314–320. 2010. View Article : Google Scholar
13	Gui D, Wei L, Jian G, Guo Y, Yang J and Wang N: Notoginsenoside R1 ameliorates podocyte adhesion under diabetic condition through α3β1 integrin upregulation in vitro and in vivo. Cell Physiol Biochem. 34:1849–1862. 2014. View Article : Google Scholar
14	Hartleben B, Gödel M, Meyer-Schwesinger C, Liu S, Ulrich T, Köbler S, Wiech T, Grahammer F, Arnold SJ, Lindenmeyer MT, et al: Autophagy influences glomerular disease susceptibility and maintains podocyte homeostasis in aging mice. J Clin Invest. 120:1084–1096. 2010. View Article : Google Scholar : PubMed/NCBI
15	Schiffer M, Bitzer M, Roberts IS, Kopp JB, ten Dijke P, Mundel P and Böttinger EP: Apoptosis in podocytes induced by TGF-beta and Smad7. J Clin Invest. 108:807–816. 2001. View Article : Google Scholar : PubMed/NCBI
16	Isermann B, Vinnikov IA, Madhusudhan T, Herzog S, Kashif M, Blautzik J, Corat MA, Zeier M, Blessing E, Oh J, et al: Activated protein C protects against diabetic nephropathy by inhibiting endothelial and podocyte apoptosis. Nat Med. 13:1349–1358. 2007. View Article : Google Scholar : PubMed/NCBI
17	Mundel P and Shankland SJ: Podocyte biology and response to injury. J Am Soc Nephrol. 13:3005–3015. 2002. View Article : Google Scholar : PubMed/NCBI
18	Zeng C, Fan Y, Wu J, Shi S, Chen Z, Zhong Y, Zhang C, Zen K and Liu Z: Podocyte autophagic activity plays a protective role in renal injury and delays the progression of podocytopathies. J Pathol. 234:203–213. 2014.PubMed/NCBI
19	Tejada T, Catanuto P, Ijaz A, Santos JV, Xia X, Sanchez P, Sanabria N, Lenz O, Elliot SJ and Fornoni A: Failure to phosphorylate AKT in podocytes from mice with early diabetic nephropathy promotes cell death. Kidney Int. 73:1385–1393. 2008. View Article : Google Scholar : PubMed/NCBI
20	Xiao H, Shi W, Liu S, Wang W, Zhang B, Zhang Y, Xu L, Liang X and Liang Y: 1,25-Dihydroxyvitamin D(3) prevents puromycin aminonucleoside-induced apoptosis of glomerular podocytes by activating the phosphatidylinositol 3-kinase/Akt-signaling pathway. Am J Nephrol. 30:34–43. 2009. View Article : Google Scholar : PubMed/NCBI
21	Feng Z, Tang L, Wu L, Cui S, Hong Q, Cai G, Wu D, Fu B, Wei R and Chen X: Na⁺/H⁺ exchanger-1 reduces podocyte injury caused by endoplasmic reticulum stress via autophagy activation. Lab Invest. 94:439–454. 2014. View Article : Google Scholar : PubMed/NCBI
22	Cinà DP, Onay T, Paltoo A, Li C, Maezawa Y, De Arteaga J, Jurisicova A and Quaggin SE: Inhibition of MTOR disrupts autophagic flux in podocytes. J Am Soc Nephrol. 23:412–420. 2012. View Article : Google Scholar :
23	Gödel M, Hartleben B, Herbach N, Liu S, Zschiedrich S, Lu S, Debreczeni-Mór A, Lindenmeyer MT, Rastaldi MP, Hartleben G, et al: Role of mTOR in podocyte function and diabetic nephropathy in humans and mice. J Clin Invest. 121:2197–2209. 2011. View Article : Google Scholar : PubMed/NCBI
24	Saleem MA, O'Hare MJ, Reiser J, Coward RJ, Inward CD, Farren T, Xing CY, Ni L, Mathieson PW and Mundel P: A conditionally immortalized human podocyte cell line demonstrating nephrin and podocin expression. J Am Soc Nephrol. 13:630–638. 2002.PubMed/NCBI
25	Li C, Chen J, Lu B, Shi Z, Wang H, Zhang B, Zhao K, Qi W, Bao J and Wang Y: Molecular switch role of Akt in Polygonatum odoratum lectin-induced apoptosis and autophagy in human non-small cell lung cancer A549 cells. PLoS One. 9:e1015262014. View Article : Google Scholar : PubMed/NCBI
26	Wang Y, Liu Y, Wang H, Li C, Qi P and Bao J: Agaricus bisporus lectins mediates islet β-cell proliferation through regulation of cell cycle proteins. Exp Biol Med (Maywood). 237:287–296. 2012. View Article : Google Scholar
27	Wang Y, Wang H, Liu Y, Li C, Qi P and Bao J: Antihyperglycemic effect of ginsenoside Rh2 by inducing islet β-cell regeneration in mice. Horm Metab Res. 44:33–40. 2012. View Article : Google Scholar
28	Rantanen M, Palmén T, Pätäri A, Ahola H, Lehtonen S, Aström E, Floss T, Vauti F, Wurst W and Ruiz P: Nephrin TRAP mice lack slit diaphragms and show fibrotic glomeruli and cystic tubular lesions. J Am Soc Nephrol. 13:1586–1594. 2002. View Article : Google Scholar : PubMed/NCBI
29	Kawachi H, Koike H, Kurihara H, Sakai T and Shimizu F: Cloning of rat homologue of podocin: expression in proteinuric states and in developing glomeruli. J Am Soc Nephrol. 14:46–56. 2003. View Article : Google Scholar
30	Wiggins JE, Goyal M, Sanden SK, Wharram BL, Shedden KA, Misek DE, Kuick RD and Wiggins RC: Podocyte hypertrophy, 'adaptation,' and 'decompensation' associated with glomerular enlargement and glomerulosclerosis in the aging rat: prevention by calorie restriction. J Am Soc Nephrol. 16:2953–2966. 2005. View Article : Google Scholar : PubMed/NCBI
31	Schmittgen TD and Livak KJ: Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 3:1101–1108. 2008. View Article : Google Scholar : PubMed/NCBI
32	Reiser J, Kriz W, Kretzler M and Mundel P: The glomerular slit diaphragm is a modified adherens junction. J Am Soc Nephrol. 11:1–8. 2000.PubMed/NCBI
33	Nielsen JS and McNagny KM: The role of podocalyxin in health and disease. J Am Soc Nephrol. 20:1669–1676. 2009. View Article : Google Scholar : PubMed/NCBI
34	Yaoita E, Kawasaki K, Yamamoto T and Kihara I: Variable expression of desmin in rat glomerular epithelial cells. The Am J Pathol. 136:899–908. 1990.PubMed/NCBI
35	Floege J, Alpers CE, Sage EH, Pritzl P, Gordon K, Johnson RJ and Couser WG: Markers of complement-dependent and complement-independent glomerular visceral epithelial cell injury in vivo. Expression of antiadhesive proteins and cytoskeletal changes. Lab Invest. 67:486–497. 1992.PubMed/NCBI
36	Somlo S and Mundel P: Getting a foothold in nephrotic syndrome. Nat Genet. 24:333–335. 2000. View Article : Google Scholar : PubMed/NCBI
37	Kerjaschki D: Caught flat-footed: podocyte damage and the molecular bases of focal glomerulosclerosis. J Clin Invest. 108:1583–1587. 2001. View Article : Google Scholar : PubMed/NCBI
38	Saleem MA, Ni L, Witherden I, Tryggvason K, Ruotsalainen V, Mundel P and Mathieson PW: Co-localization of nephrin, podocin, and the actin cytoskeleton: evidence for a role in podocyte foot process formation. Am J Pathol. 161:1459–1466. 2002. View Article : Google Scholar : PubMed/NCBI
39	Asanuma K, Tanida I, Shirato I, Ueno T, Takahara H, Nishitani T, Kominami E and Tomino Y: MAP-LC3, a promising autophagosomal marker, is processed during the differentiation and recovery of podocytes from PAN nephrosis. FASEB J. 17:1165–1167. 2003.PubMed/NCBI
40	Nunez G, Benedict MA, Hu Y and Inohara N: Caspases: the proteases of the apoptotic pathway. Oncogene. 17:3237–3245. 1998. View Article : Google Scholar
41	Mohr S, McCormick TS and Lapetina EG: Macrophages resistant to endogenously generated nitric oxide-mediated apoptosis are hypersensitive to exogenously added nitric oxide donors: dichotomous apoptotic response independent of caspase 3 and reversal by the mitogen-activated protein kinase kinase (MEK) inhibitor PD 098059. Proc Natl Acad Sci USA. 95:5045–5050. 1998. View Article : Google Scholar
42	Wada T, Pippin JW, Marshall CB, Griffin SV and Shankland SJ: Dexamethasone prevents podocyte apoptosis induced by puromycin aminonucleoside: role of p53 and Bcl-2-related family proteins. J Am Soc Nephrol. 16:2615–2625. 2005. View Article : Google Scholar : PubMed/NCBI
43	Li X, Zhang X, Li X, Ding F and Ding J: The role of survivin in podocyte injury induced by puromycin aminonucleoside. Int J Mol Sci. 15:6657–6673. 2014. View Article : Google Scholar : PubMed/NCBI
44	Chen YQ, Wang XX, Yao XM, Zhang DL, Yang XF, Tian SF and Wang NS: MicroRNA-195 promotes apoptosis in mouse podocytes via enhanced caspase activity driven by BCL2 insufficiency. Am J Nephrol. 34:549–559. 2011. View Article : Google Scholar : PubMed/NCBI
45	Gui D, Guo Y, Wang F, Liu W, Chen J, Chen Y, Huang J and Wang N: Astragaloside IV, a novel antioxidant, prevents glucose-induced podocyte apoptosis in vitro and in vivo. PLoS One. 7:e398242012. View Article : Google Scholar : PubMed/NCBI
46	Cantley LC: The phosphoinositide 3-kinase pathway. Science. 296:1655–1657. 2002. View Article : Google Scholar : PubMed/NCBI
47	Franke TF, Hornik CP, Segev L, Shostak GA and Sugimoto C: PI3K/Akt and apoptosis: size matters. Oncogene. 22:8983–8998. 2003. View Article : Google Scholar : PubMed/NCBI
48	Huber TB, Hartleben B, Kim J, Schmidts M, Schermer B, Keil A, Egger L, Lecha RL, Borner C, Pavenstädt H, et al: Nephrin and CD2AP associate with phosphoinositide 3-OH kinase and stimulate AKT-dependent signaling. Mol Cell Biol. 23:4917–4928. 2003. View Article : Google Scholar : PubMed/NCBI