CN: 32-1845/R
ISSN: 2095-6975
Cite this paper:
LI Yun-Ying, LU Xiao-Yan, SUN Jia-Li, WANG Qing-Qing, ZHANG Yao-Dan, ZHANG Jian-Bing, FAN Xiao-Hui. Potential hepatic and renal toxicity induced by the biflavonoids from Ginkgo biloba[J]. Chinese Journal of Natural Medicines, 2019, 17(9): 672-681

Potential hepatic and renal toxicity induced by the biflavonoids from Ginkgo biloba

LI Yun-Ying1, LU Xiao-Yan2, SUN Jia-Li2, WANG Qing-Qing3, ZHANG Yao-Dan3, ZHANG Jian-Bing3, FAN Xiao-Hui1,2
1 Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China;
2 Pharmaceutical Informatics Institute, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China;
3 Zhejiang University-Wanbangde Pharmaceutical Group Joint Research Center for Chinese Medicine Modernization, Hangzhou 310058, China
Evidence continues to grow on potential health risks associated with Ginkgo biloba and its constituents. While biflavonoid is a subclass of the flavonoid family in Ginkgo biloba with a plenty of pharmacological properties, the potential toxicological effects of biflavonoids remains largely unknown. Thus, the aim of this study was to investigate the in vitro and in vivo toxicological effects of the biflavonoids from Ginkgo biloba (i.e., amentoflavone, sciadopitysin, ginkgetin, isoginkgetin, and bilobetin). In the in vitro cytotoxicity test, the five biflavonoids all reduced cell viability in a dose-dependent manner in human renal tubular epithelial cells (HK-2) and human normal hepatocytes (L-02), indicating they might have potential liver and kidney toxicity. In the in vivo experi-ments, after intragastrical administration of these biflavonoids at 20 mg·kg-1·d-1 for 7 days, serum biochemical analysis and histo-pathological examinations were performed. The activity of alkaline phosphatase was significantly increased after all the biflavonoid administrations and widespread hydropic degeneration of hepatocytes was observed in ginkgetin or bilobetin-treated mice. More-over, the five biflavonoids all induced acute kidney injury in treated mice and the main pathological lesions were confirmed to the tubule, glomeruli, and interstitium injuries. As the in vitro and in vivo results suggested that these biflavonoids may be more toxic to the kidney than the liver, we further detected the mechanism of biflavonoids-induced nephrotoxicity. The increased TUNEL-positive cells were detected in kidney tissues of biflavonoids-treated mice, accompanied by elevated expression of proapoptotic protein BAX and unchanged levels of antiapoptotic protein BCL-2, indicating apoptosis was involved in biflavonoids-induced nephrotoxicity. Taken together, our results suggested that the five biflavonoids from Ginkgo biloba may have potential hepatic and renal toxicity and more attentions should be paid to ensure Ginkgo biloba preparations safety.
Key words:    Biflavonoids    Ginkgo biloba    Potential toxicity    Liver    Kidney    Apoptosis   
Received: 2019-05-17   Revised:
PDF (14435 KB) Free
Print this page
Email this article to others
Articles by LI Yun-Ying
Articles by LU Xiao-Yan
Articles by SUN Jia-Li
Articles by WANG Qing-Qing
Articles by ZHANG Yao-Dan
Articles by ZHANG Jian-Bing
Articles by FAN Xiao-Hui
[1] Yao X, Chen N, Ma CH, et al. Ginkgo biloba extracts attenuate lipopolysaccharide-induced inflammatory responses in acute lung injury by inhibiting the COX-2 and NF-κB pathways[J]. Chin J Nat Med, 2015, 13(1):52-58.
[2] Ahlemeyer B, Krieglstein J. Pharmacological studies supporting the therapeutic use of Ginkgo biloba extract for Alzheimer's disease[J]. Pharmacopsychiatry, 2003, 36(S1):8-14.
[3] Pietri S, Maurelli E, Drieu E, et al. Cardioprotective and anti-oxidant effects of the terpenoid constituents of Ginkgo biloba extract (EGb 761)[J]. J Mol Cell Cardiol, 1997, 29(2):733-742.
[4] Gohil K, Moy RK, Farzin S, et al. mRNA expression profile of a human cancer cell line in response to Ginkgo biloba extract:induction of antioxidant response and the Golgi system[J]. Free Radic Res, 2000, 33(6):831-849.
[5] Rhee KJ, Lee CG, Kim SW, et al. Extract of Ginkgo biloba ameliorates Streptozotocin-induced type 1 diabetes mellitus and high-fat diet-induced type 2 diabetes mellitus in mice[J]. Int J Med Sci, 2015, 12(12):987-994.
[6] Zha WB, A JY, Wang GJ, et al. Metabonomic approach to evaluating pharmacodynamics of Ginkgo biloba extract on the perturbed metabolism in hamsters with atherosclerosis by high fat diet[J]. Chin J Nat Med, 2011, 9(3):232-240.
[7] Mei N, Guo X, Ren Z, et al. Review of Ginkgo biloba-induced toxicity, from experimental studies to human case reports[J]. J Environ Sci Health C Environ Carcinog Ecotoxicol Rev, 2017, 35(1):1-28.
[8] Jacobs BP, Browner WS. Ginkgo biloba:a living fossil[J]. Am J Med, 2000, 108(4):341-342.
[9] Lu XY, Chen L, Liu T, et al. Chemical analysis, pharmacological activity and process optimization of the proportion of bilobalide and ginkgolides in Ginkgo biloba extract[J]. J Pharm Biomed Anal, 2018, 160:46-54.
[10] Smith JV, Luo Y. Studies on molecular mechanisms of Ginkgo biloba extract[J]. Appl Microbiol Biotechnol, 2004, 64(4):465-472.
[11] Shan SJ, Zhang PP, Luo J, et al. Two new phenolic glycosides isolated from Ginkgo seeds[J]. Chin J Nat Medicines, 2018, 16(7):505-508.
[12] Gontijo VS, Dos Santos MH, Viegas C Jr. Biological and chemical aspects of natural biflavonoids from plants:a brief review[J]. Mini Rev Med Chem, 2017, 17(10):834-862.
[13] Ishola IO, Chaturvedi JP, Rai S, et al. Evaluation of amen-toflavone isolated from Cnestis ferruginea Vahl ex DC (Connaraceae) on production of inflammatory mediators in LPS stimulated rat astrocytoma cell line (C6) and THP-1 cells[J]. J Ethnopharmacol, 2013, 146(2):440-448.
[14] Miki K, Nagai T, Suzuki K, et al. Anti-influenza virus activity of biflavonoids[J]. Bioorg Med Chem Lett, 2007, 17(3):772-775.
[15] Cao J, Tong C, Liu Y, et al. Ginkgetin inhibits growth of breast carcinoma via regulating MAPKs pathway[J]. Biomed Pharmacother, 2017, 96:450-458.
[16] Lee JS, Lee MS, Oh WK, et al. Fatty acid synthase inhibition by amentoflavone induces apoptosis and antiproliferation in human breast cancer cells[J]. Biol Pharm Bull, 2009, 32(8):1427-1432.
[17] Laishram S, Sheikh Y, Moirangthem DS, et al. Anti-diabetic molecules from Cycas pectinata Griff. traditionally used by the Maiba-Maibi[J]. Phytomedicine, 2015, 22(1):23-26.
[18] Son JK, Son MJ, Lee E, et al. Ginkgetin, a biflavone from Ginko biloba leaves, inhibits cyclooxygenases-2 and 5-lipoxygenase in mouse bone marrowderived mast cells[J]. Biol Pharm Bull, 2005, 28(12):2181-2184.
[19] Miao LP, Wang XJ, Zhou HF, et al. Advance on the pharmacological effects of biflavonoids[J]. World Clinical Drugs, 2012, 33(6):369-375.
[20] Chen JW, Xiang L, Ping X, et al. Application of sciadopitysin in the preparation of diabetes drugs[P]. CN201310032630.2(2013).
[21] Yang Z, Zhu J, Zhang H, et al. Investigating chemical features of Panax notoginseng based on integrating HPLC fingerprinting and determination of multiconstituents by single reference standard[J]. J Gins Res, 2018, 42(3):334-342.
[22] Zheng J, Yu LQ, Chen W, et al. Circulating exosomal mi-croRNAs reveal the mechanism of Fructus Meliae Toosendan-induced liver injury in mice[J]. Sci Rep, 2018, 8(1):2832.
[23] Lu X, Ji C, Tong W, et al. Integrated analysis of microRNA and mRNA expression profiles highlights the complex and dynamic behavior of toosendanin-induced liver injury in mice[J]. Sci Rep, 2016, 6:34225.
[24] Ji C, Zheng J, Tong W, et al. Revealing the mechanism of melia toosendan fruit-induced liver injury in mice by integrating microRNA and mRNA-Based toxicogenomics data[J]. RSC Adv, 2015, 10:1039.
[25] Zheng J, Ji C, Lu X, et al. Integrated expression profiles of mRNA and microRNA in the liver of Fructus Meliae Toosendan water extract injured mice[J]. Front Pharmacol, 2015, 6:236.
[26] Baron-Ruppert G, Luepke NP. Evidence for toxic effects of alkylphenols from Ginkgo biloba in the hen's egg test (HET)[J]. Phytomedicine, 2001, 8(2):133-138.
[27] Lin JL, Ho YS. Flavonoid-induced acute nephropathy[J]. Am J Kidney Dis, 1994, 23(3):433-440.
[28] Kimura Y, Ito H, Ohnishi R, et al. Inhibitory effects of polyphenols on human cytochrome P4503A4 and 2C9 activity[J]. Food Chem Toxicol, 2010, 48(1):429-435.
[29] Cardoso CR, de Syllos Colus IM, Bernardi CC, et al. Mutagenic activity promoted by amentoflavone and methanolic extract of Byrsonima crassa Niedenzu[J]. Toxicology, 2006, 225(1):55-63.
[30] Lv X, Zhang JB, Wang XX, et al. Amentoflavone is a potent broad-spectrum inhibitor of human UDP-glucuronosyltransferases[J]. Chem Biol Interact, 2018, 284:48-55.
[31] Liu XY, Lv X, Wang P, et al. Inhibition of UGT1A1 by natural and synthetic flavonoids[J]. Int J Biol Macromol, 2019, 126:653-661.
[32] Wang XX, Hou J, Ning J, et al. Inhibition of sciadopitysin against UDP-glucuronosyltransferases[J]. Acta Pharm Sin, 2016, 51(5):749-55.
[33] Cao J, Lu Q, Liu N, et al. Sciadopitysin suppresses RANKL-mediated osteoclastogenesis and prevents bone loss in LPS-treated mice[J]. Int Immunopharmacol, 2017, 49:109-117.
[34] Cao Q, Qin L, Huang F, et al. Amentoflavone protects dopaminergic neurons in MPTP-induced Parkinson's disease model mice through PI3K/Akt and ERK signaling pathways[J]. Toxicol Appl Pharmacol, 2017, 319:80-90.
[35] Zhang J, Yang S, Chen F, et al. Ginkgetin aglycone ameliorates LPS-induced acute kidney injury by activating SIRT1 via inhibiting the NF-κB signaling pathway[J]. Cell Biosci, 2017, 7(1):44.
[36] Kunert O, Swamy RC, Kaiser M, et al. Antiplasmodial and leishmanicidal activity of biflavonoids from Indian Se-laginella bryopteris[J]. Phytochem Lett, 2008, 1(4):171-174.
[37] Ryan MJ, Johnson G, Kirk J, et al. HK-2:an immortalized proximal tubule epithelial cell line from normal adult human kidney[J]. Kidney Int, 1994, 45(1):48-57.
[38] Murphy RA, Stafford RM, Petrasovits BA, et al. Estab-lishment of HK-2 cells as a relevant model to study tenofovir-induced cytotoxicity[J]. Int J Mol Sci, 2017, 18(3):531-533.
[39] Cheng L, Ge M, Lan Z, et al. Zoledronate dysregulates fatty acid metabolism in renal tubular epithelial cells to induce nephrotoxicity[J]. Arch Toxicol, 2018, 92(1):469-485.
[40] Gunness P, Aleksa K, Kosuge K, et al. Comparison of the novel HK-2 human renal proximal tubular cell line with the standard LLC-PK1 cell line in studying drug-induced nephrotoxicity[J]. Can J Physiol Pharmacol, 2010, 88(4):448-455.
[41] Hu X, Yang Y, Li C, et al. Human fetal hepatocyte line, L-02, exhibits good liver function in vitro and in an acute liver failure model[J]. Transplant Proc, 2013, 45(2):695-700.
[42] Xiao Y, Zeng M, Yin L, et al. Clusterin increases mito-chondrial respiratory chain complex I activity and protects against hexavalent chromium-induced cytotoxicity in L-02 hepatocytes[J]. Toxicol Res (Camb), 2019, 8(1):15-24.
[43] Liang Q, Xiao Y, Liu K, et al. Cr(VI)-induced autophagy protects L-02 hepatocytes from apoptosis through the ROS-AKT-mTOR pathway[J]. Cell Physiol Biochem, 2018, 51(4):1863-1878.
[44] Nicolson TJ, Mellor HR, Roberts RRA. Gender differ-ences in drug toxicity[J]. Trends Pharmacol Sci, 2010, 31(3):0-114.
[45] Buzzetti E, Parikh PM, Gerussi A, et al. Gender differ-ences in liver disease and the drug-dose gender gap[J]. Pharmacol Res, 2017, 120:97-108.
[46] Shi Q, Hong H, Senior J, et al. Biomarkers for drug-induced liver injury[J]. Expert Rev Gastroent, 2010, 4(2):225.
[47] Lala V, Minter DA. Liver Function Tests[M]. StatPearls. 2019.
[48] Hirschfield GM. Diagnosis of primary biliary cirrhosis[J]. Best Pract Res Clin Gastroenterol, 2011, 25(6):701-712.
[49] Elevated Alkaline Phosphatase-Levels, Causes and Treatment[M]. 2016.
[50] Pushpavalli G, Veeramani C, Pugalendi KV. Influence of chrysin on hepatic marker enzymes and lipid profile against D-galactosamine-induced hepatotoxicity rats[J]. Food Chem Toxicol, 2010, 48(6):1654-1659.
[51] Wang Y, Jiang ZZ, Chen M, et al. Protective effect of total flavonoid C-glycosides from Abrus mollis extract on lipopolysaccharide-induced lipotoxicity in mice[J]. Chin J Nat Med, 2014, 12(6):461-468.
[52] Gobe GC, Coombes JS, Fassett RG, et al. Biomarkers of drug-induced acute kidney injury in the adult[J]. Expert Opin Drug Metab Toxicol, 2015, 11(11):1683-1694.
[53] Endre ZH, Pickering JW, Walker RJ. Clearance and beyond:the complementary roles of GFR measurement and injury biomarkers in acute kidney injury (AKI)[J]. Am J Physiol Renal Physiol, 2011, 301(4):697-707.
[54] Yan M, Tang C, Ma Z, et al. DNA damage response in nephrotoxic and ischemic kidney injury[J]. Toxicol Appl Pharmacol, 2016, 313:104-108.
[55] Zhu S, Pabla N, Tang C, et al. DNA damage response in cisplatin-induced nephrotoxicity[J]. Arch Toxicol, 2015, 89(12):2197-2205.
[56] Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in-vivo with a conserved homolog, bax, that accelerates programmed cell-death[J]. Cell, 1993, 74(4):609-619.
[57] Czabotar PE, Lessene G, Strasser A, et al. Control of apoptosis by the BCL-2 protein family:implications for physiology and therapy[J]. Nat Rev Mol Cell Biol, 2014, 15(1):49-63.

Related Articles:
1. EZEJIOFOR Anthoneth Ndidi, ORISH Chinna Nneka, ORISAKWE Orish Ebere.Cytological and biochemical studies during the progression of alloxan-induced diabetes and possible protection of an aqueous leaf extract of Costus afer[J]. Chinese Journal of Natural Medicines, 2014,12(10): 745-752