<em>Ginkgo biloba</em> leaf extract prevents diabetic nephropathy through the suppression of tissue transglutaminase

Yu,Xiaoyan; Su,Qing; Geng,Jianan; Liu,Hui; Liu,Yumeng; Liu,Jinming; Shi,Yan; Zou,Yinggang

doi:10.3892/etm.2021.9764

Ginkgo biloba leaf extract prevents diabetic nephropathy through the suppression of tissue transglutaminase

Authors:
- Xiaoyan Yu
- Qing Su
- Jianan Geng
- Hui Liu
- Yumeng Liu
- Jinming Liu
- Yan Shi
- Yinggang Zou
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Affiliations: Department of Experimental Pharmacology and Toxicology, School of Pharmaceutical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China, Department of Anatomy, College of Basic Medical Science, Jilin University, Changchun, Jilin 130012, P.R. China, Department of Obstetrics and Gynecology, The Second Hospital, Jilin University, Changchun, Jilin 130041, P.R. China
Published online on: February 8, 2021 https://doi.org/10.3892/etm.2021.9764
Article Number: 333
Copyright: © Yu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to investigate the preventive effects of Ginkgo biloba leaf extract (GBE) against extracellular matrix (ECM) accumulation in a streptozotocin (STZ)‑induced rat model of diabetic nephropathy (DN), and to determine its underlying molecular mechanism. In vivo, a rat model of DN was established by intraperitoneal injection of STZ, and the rats were subsequently administered GBE. The results demonstrated that GBE significantly decreased blood glucose, the urine protein excretion rate and ECM accumulation in DN rats. In addition, the development of DN significantly induced tissue transglutaminase (tTG) protein expression, which was detected by immunohistochemistry, western blotting and PCR analyses, while GBE administration decreased tTG expression in the diabetic kidney. In vitro, rat glomerular mesangial cells (HBZY‑1 cells) cultured with high glucose were also treated with GBE. The concentrations of tTG, fibronectin, type IV collagen, transforming growth factor (TGF)‑β and connective tissue growth factor (CTGF) were detected via ELISA. The results demonstrated that GBE notably decreased the concentration of these proteins, and tTG expression was positively associated with TGF‑β. GBE also suppressed tTG expression of high glucose‑treated HBZY‑1 cells in a concentration‑dependent manner. Furthermore, tTG protein expression was detected in high glucose‑treated HBZY‑1 cells transfected with small interfering RNA (siRNA) oligonucleotides against TGF‑β and CTGF to investigate a possible mechanism of GBE‑mediated inhibition of tTG. The results demonstrated that the tTG levels remained unchanged in CTGF siRNA‑transfected cells, but were decreased in the GBE + CTGF siRNA group compared with the control siRNA group, suggesting that tTG may not be regulated by CTGF, and the inhibitory effect of GBE on tTG may not be associated with the direct inhibition of CTGF. However, tTG expression was decreased following the transfection with TGF‑β siRNA, in which levels of tTG were similar compared with both the GBE group and GBE + TGF‑β siRNA group, indicating that tTG may be regulated by TGF‑β, and that the GBE‑induced repression of tTG expression may be associated with the downregulation of TGF‑β. Taken together, the results of the present study suggest that GBE prevented ECM accumulation by suppressing tTG expression in DN, which was predominantly mediated by TGF‑β.

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1	Guthrie RA and Guthrie DW: Pathophysiology of diabetes mellitus. Crit Care Nurs Q. 27:113–125. 2004.PubMed/NCBI View Article : Google Scholar
2	Islam MA, Amin MN, Siddiqui SA, Hossain MP, Sultana F and Kabir MR: Trans fatty acids and lipid profile: A serious risk factor to cardiovascular disease, cancer and diabetes. Diabetes Metab Syndr. 13:1643–1647. 2019.PubMed/NCBI View Article : Google Scholar
3	Schmidt AM: Highlighting diabetes mellitus: The epidemic continues. Arterioscler Thromb Vasc Biol. 38:e1–e8. 2018.PubMed/NCBI View Article : Google Scholar
4	Tung CW, Hsu YC, Shih YH, Chang PJ and Lin CL: Glomerular mesangial cell and podocyte injuries in diabetic nephropathy. Nephrology (Carlton). 23 (Suppl 4):S32–S37. 2018.PubMed/NCBI View Article : Google Scholar
5	Schelling JR: Tissue transglutaminase inhibition as treatment for diabetic glomerular scarring: It's good to be glueless. Kidney Int. 76:363–365. 2009.PubMed/NCBI View Article : Google Scholar
6	Skill NJ, Johnson TS, Coutts IG, Saint RE, Fisher M, Huang L, El Nahas AM, Collighan RJ and Griffin M: Inhibition of transglutaminase activity reduces extracellular matrix accumulation induced by high glucose levels in proximal tubular epithelial cells. J Biol Chem. 279:47754–47762. 2004.PubMed/NCBI View Article : Google Scholar
7	Fleckenstein B, Qiao SW, Larsen MR, Jung G, Roepstorff P and Sollid LM: Molecular characterization of covalent complexes between tissue transglutaminase and gliadin peptides. J Biol Chem. 279:17607–17616. 2004.PubMed/NCBI View Article : Google Scholar
8	El Nahas AM, Abo-Zenah H, Skill NJ, Bex S, Wild G, Griffin M and Johnson TS: Elevated epsilon-(gamma-glutamyl)lysine in human diabetic nephropathy results from increased expression and cellular release of tissue transglutaminase. Nephron Clin Pract. 97:c108–c117. 2004.PubMed/NCBI View Article : Google Scholar
9	Skill NJ, Griffin M, El Nahas AM, Sanai T, Haylor JL, Fisher M, Jamie MF, Mould NN and Johnson TS: Increases in renal epsilon-(gamma-glutamyl)-lysine crosslinks result from compartment-specific changes in tissue transglutaminase in early experimental diabetic nephropathy: Pathologic implications. Lab Invest. 81:705–716. 2001.PubMed/NCBI View Article : Google Scholar
10	Loeffler I and Wolf G: Transforming growth factor-β and the progression of renal disease. Nephrol Dial Transplant. 29 (Suppl 1):i37–i45. 2014.PubMed/NCBI View Article : Google Scholar
11	Border WA and Noble NA: Evidence that TGF-beta should be a therapeutic target in diabetic nephropathy. Kidney Int. 54:1390–1391. 1998.PubMed/NCBI View Article : Google Scholar
12	Sutariya B, Jhonsa D and Saraf MN: TGF-β: The connecting link between nephropathy and fibrosis. Immunopharmacol Immunotoxicol. 38:39–49. 2016.PubMed/NCBI View Article : Google Scholar
13	Lee HS: Paracrine role for TGF-β-induced CTGF and VEGF in mesangial matrix expansion in progressive glomerular disease. Histol Histopathol. 27:1131–1141. 2012.PubMed/NCBI View Article : Google Scholar
14	Roestenberg P, van Nieuwenhoven FA, Wieten L, Boer P, Diekman T, Tiller AM, Wiersinga WM, Oliver N, Usinger W, Weitz S, et al: Connective tissue growth factor is increased in plasma of type 1 diabetic patients with nephropathy. Diabetes Care. 27:1164–1170. 2004.PubMed/NCBI View Article : Google Scholar
15	Wahab NA, Yevdokimova N, Weston BS, Roberts T, Li XJ, Brinkman H and Mason RM: Role of connective tissue growth factor in the pathogenesis of diabetic nephropathy. Biochem J. 359:77–87. 2001.PubMed/NCBI View Article : Google Scholar
16	Rosenbloom J, Ren S and Macarak E: New frontiers in fibrotic disease therapies: The focus of the Joan and Joel rosenbloom center for fibrotic diseases at thomas jefferson university. Matrix Biol. 51:14–25. 2016.PubMed/NCBI View Article : Google Scholar
17	Lu Q, Yin XX, Wang JY, Gao YY and Pan YM: Effects of Ginkgo biloba on prevention of development of experimental diabetic nephropathy in rats. Acta Pharmacol Sin. 28:818–828. 2007.PubMed/NCBI View Article : Google Scholar
18	Ji L, Yin XX, Wu ZM, Wang JY, Lu Q and Gao YY: Ginkgo biloba extract prevents glucose-induced accumulation of ECM in rat mesangial cells. Phytother Res. 23:477–485. 2009.PubMed/NCBI View Article : Google Scholar
19	Wang D, Li X, Yu X, Shi Y and Yin L: Expression of tissue transglutaminase on renal interstitial fibrosis rats and intervention of GBE. Zhongguo Zhong Yao Za Zhi. 34:1133–1136. 2009.PubMed/NCBI(In Chinese).
20	Chinese Pharmacopoeia Commission. Pharmacopoeia of the People's Republic of China. Vol. 1. Beijing. China Medical Science and Technology Press, 2015.
21	Yang M, Kan L, Wu L, Zhu Y and Wang Q: Effect of baicalin on renal function in patients with diabetic nephropathy and its therapeutic mechanism. Exp Ther Med. 17:2071–2076. 2019.PubMed/NCBI View Article : Google Scholar
22	Afghahi H, Svensson MK, Pirouzifard M, Eliasson B and Svensson AM: Blood pressure level and risk of major cardiovascular events and all-cause of mortality in patients with type 2 diabetes and renal impairment: An observational study from the swedish national diabetes register. Diabetologia. 58:1203–1211. 2015.PubMed/NCBI View Article : Google Scholar
23	Rogers NM, Teubner DJ and Coates PT: Calcific uremic arteriolopathy: Advances in pathogenesis and treatment. Semin Dial. 20:150–157. 2007.PubMed/NCBI View Article : Google Scholar
24	Eberhardt W, Engels C, Müller R and Pfeilschifter J: Mechanisms of dexamethasone-mediated inhibition of cAMP-induced tPA expression in rat mesangial cells. Kidney Int. 62:809–821. 2002.PubMed/NCBI View Article : Google Scholar
25	Fang J, Wang Z, Wang P and Wang M: Extraction, structure and bioactivities of the polysaccharides from Ginkgo biloba: A review. Int J Biol Macromol. 162:1897–1905. 2020.PubMed/NCBI View Article : Google Scholar
26	Castro NE, Kato M, Park JT and Natarajan R: Transforming growth factor β1 (TGF-β1) enhances expression of profibrotic genes through a novel signaling cascade and microRNAs in renal mesangial cells. J Biol Chem. 289:29001–29013. 2014.PubMed/NCBI View Article : Google Scholar
27	Solini A, Manca ML, Penno G, Pugliese G, Cobb JE and Ferrannini E: Prediction of declining renal function and albuminuria in patients with type 2 diabetes by metabolomics. J Clin Endocrinol Metab. 101:696–704. 2016.PubMed/NCBI View Article : Google Scholar
28	Tian J, Popal MS and Liu Y, Gao R, Lyu S, Chen K and Liu Y: Ginkgo biloba leaf extract attenuates atherosclerosis in streptozotocin-induced diabetic ApoE^-/- mice by inhibiting endoplasmic reticulum stress via restoration of autophagy through the mTOR signaling pathway. Oxid Med Cell Longev. 2019(8134678)2019.PubMed/NCBI View Article : Google Scholar
29	Coskun O, Armutcu F, Kanter M and Kuzey GM: Protection of endotoxin-induced oxidative renal tissue damage of rats by vitamin E or/and EGb 761 treatment. J Appl Toxicol. 25:8–12. 2005.PubMed/NCBI View Article : Google Scholar
30	Lasaite L, Spadiene A, Savickiene N, Skesters A and Silova A: The effect of Ginkgo biloba and Camellia sinensis extracts on psychological state and glycemic control in patients with type 2 diabetes mellitus. Nat Prod Commun. 9:1345–1350. 2014.PubMed/NCBI
31	Cui JF, Yang W, Xie YM, Sun Y, Zhuang Y and Wang YY: Real-world analysis of concurrent diseases and medicine use among patients with insomnia. Zhongguo Zhong Yao Za Zhi. 39:3519–3526. 2014.PubMed/NCBI(In Chinese).
32	Pohlers D, Brenmoehl J, Löffler I, Müller CK, Leipner C, Schultze-Mosgau S, Stallmach A, Kinne RW and Wolf G: TGF-beta and fibrosis in different organs-molecular pathway imprints. Biochim Biophys Acta. 1792:746–756. 2009.PubMed/NCBI View Article : Google Scholar
33	Serban AI, Stanca L, Geicu OI, Munteanu MC and Dinischiotu A: RAGE and TGF-β1 cross-talk regulate extracellular matrix turnover and cytokine synthesis in AGEs exposed fibroblast cells. PLoS One. 11(e0152376)2016.PubMed/NCBI View Article : Google Scholar
34	Cui Y, Robertson J, Maharaj S, Waldhauser L, Niu J, Wang J, Farkas L, Kolb M and Gauldie J: Oxidative stress contributes to the induction and persistence of TGF-β1 induced pulmonary fibrosis. Int J Biochem Cell Biol. 43:1122–1133. 2011.PubMed/NCBI View Article : Google Scholar
35	Yin Q and Liu H: Connective tissue growth factor and renal fibrosis. Adv Exp Med Biol. 1165:365–380. 2019.PubMed/NCBI View Article : Google Scholar
36	Douthwaite JA, Johnson TS, Haylor JL, Watson P and El Nahas AM: Effects of transforming growth factor-beta1 on renal extracellular matrix components and their regulating proteins. J Am Soc Nephrol. 10:2109–2119. 1999.PubMed/NCBI
37	Akimov SS and Belkin AM: Cell-surface transglutaminase promotes fibronectin assembly via interaction with the gelatin-binding domain of fibronectin: A role in TGFbeta-dependent matrix deposition. J Cell Sci. 114:2989–3000. 2001.PubMed/NCBI
38	Zhang H, Cai X, Yi B, Huang J, Wang J and Sun J: Correlation of CTGF gene promoter methylation with CTGF expression in type 2 diabetes mellitus with or without nephropathy. Mol Med Rep. 9:2138–2144. 2014.PubMed/NCBI View Article : Google Scholar
39	Hou X, Tian J, Geng J, Li X, Tang X, Zhang J and Bai X: MicroRNA-27a promotes renal tubulointerstitial fibrosis via suppressing PPARγ pathway in diabetic nephropathy. Oncotarget. 7:47760–47776. 2016.PubMed/NCBI View Article : Google Scholar
40	Tang D, Zhang Z, Gao Y, Wei Y and Han L: Protective effects of serum containing Ginkgo biloba extract on glomerulosclerosis in rat mesangial cells. J Ethnopharmacol. 124:26–33. 2009.PubMed/NCBI View Article : Google Scholar