Skip to main content

Advertisement

SpringerLink
Log in

Isolation, characterization, and therapeutic activity of bergenin from marlberry (Ardisia colorata Roxb.) leaf on diabetic testicular complications in Wistar albino rats

  • Research Article
  • Published:
Environmental Science and Pollution Research Aims and scope Submit manuscript

Abstract

Bergenin is one of the phytochemical constituents in marlberry (Ardisia colorata Roxb.) having antioxidant, anti-diabetic, and anti-inflammatory properties. A. colorata has been used as an herbal medicine in Southeast Asia particularly in Northeast India to treat diabetes. Bergenin was isolated from methanol extract of A. colorata leaf (MEACL) by column chromatography and TLC profiling. Characterization and structural validation of bergenin were performed by spectroscopic analyses. A LC-ESI-MS/MS method was developed for the quantitation of bergenin and validated as per the guidelines of FDA and EMA. The validated method was successfully utilized to quantify bergenin concentration in MEACL samples. Therapeutic efficacy of bergenin was investigated on streptozotocin-induced diabetic rats by following standard protocols. Bergenin supplementation significantly improved the physiological and metabolic processes and in turn reverses diabetic testicular dysfunction via increasing serum testosterone concentrations and expression pattern of PCNA, improving histopathological and histomorphometric manifestations, modulating spermatogenic events and germ cell proliferation, restoring sperm quality, reducing sperm DNA damage, and balancing the antioxidant enzymes levels. Hence, A. colorata leaf is one of the alternate rich resources of bergenin and could be used as a therapeutic agent for diabetic testicular complications.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

Abbreviations

B100, B200, and B400:

Diabetic rats treated with 100, 200, and 400 mg/kg of bergenin isolated from Ardisia colorata methanol leaf extract, respectively

AC:

Ardisia colorata

ALT:

Alanine aminotransferase

AO+ :

Abnormal DNA fluoresce, yellow or red

AO− :

Normal DNA fluoresce, green

AST:

Aspartate aminotransferase

C:

Healthy untreated normal control (0.5% carboxy methyl cellulose)

CMC:

Carboxy methyl cellulose

CPCSEA:

Committee for the Purpose of Control and Supervision of Experiments on Animals

DC:

Diabetic control (streptozotocin, 60 mg/kg)

DCA:

Detrended correspondence analysis

DEPT:

Distortedness enhancement by polarization transfer

DFI:

DNA fragmentation index

DM:

Diabetes mellitus

DSP:

Daily sperm production

DSPr:

Daily sperm production relative to testis weight

EMA:

European Medicine Agency

FDA:

Food and Drug Agency

FPG:

Fasting plasma glucose

GC:

Diabetic rats treated with glibenclamide (0.1 mg/kg)

GST:

Glutathione S-transferase

HPLC:

High-performance liquid chromatography

JTBS:

Johnsen’s testicular biopsy score

LC:

Liquid chromatography grade

LC-ESI-MS/MS:

Liquid chromatography and electrospray ionization mass spectrometry

LOD:

Limit of detection

LOQ:

Limit of quantification

MANOVA:

Multivariate analysis of variance

MDA:

Malondialdehyde equivalents

MEACL:

Methanol extract of Ardisia colorata leaf

MRM:

Multiple-reaction monitoring

MSTD:

Mean seminiferous tubule diameter

MZUAEC:

Mizoram University Animal Ethical Committee

OECD:

Organization for Economic Co-operation and Development

PCA:

Principal component analysis

PCNA:

Proliferating cell nuclear antigen

QC:

Quality control

ROS:

Reactive oxygen species

RSD:

Relative standard deviation

SD:

Standard deviation

SOD:

Superoxide dismutase

STZ:

Streptozotocin

TDI:

Tubule differentiation index

TLC:

Thin layer chromatography

References

  • Abdelali A, Al-Bader M, Kilarkaje N (2016) Effects of trans-resveratrol on hyperglycemia induced abnormal spermatogenesis, DNA damage and alterations in poly (ADP-ribose) polymerase signaling in rat testis. Toxicol Appl Pharmacol 311:61–73

    Article  CAS  Google Scholar 

  • Aguirre-Arias MV, Velarde V, Moreno RD (2017) Effects of ascorbic acid on spermatogenesis and sperm parameters in diabetic rats. Cell Tissue Res 370(2):305–317

    Article  CAS  Google Scholar 

  • Altay B, Evki S, Anavsargil BD, Hekimgil M, Semerci B (2003) Streptozotocin-induced diabetic effects on spermatogenesis with proliferative cell nuclear antigen immune staining of adult rat testis. Fertil Steril 80(2):1523

    Google Scholar 

  • Alves MG, Martins AD, Rato L, Moreira PI, Socorro S, Oliveira PF (2013) Molecular mechanisms beyond glucose transport in diabetes-related male infertility. Biochim Biophys Acta 1832:626–635

    Article  CAS  Google Scholar 

  • Ambika S, Saravanan R (2016) Effect of bergenin on hepatic glucose metabolism and insulin signaling in C57BL/6J mice with high fat-diet induced type 2 diabetes. J Appl Biomed 14:221–227

    Article  Google Scholar 

  • American Diabetes Association (2018) Classification and diagnosis of diabetes: standards of medical care in diabetes—2018. Diabetes Care 41(Supplement 1):S13–S27

    Article  Google Scholar 

  • Asada K, Takahashi M, Nagate M (1974) Assay and inhibitors of spinach superoxide dismutase. Agric Biol Chem 38:471–473

    Article  CAS  Google Scholar 

  • Ashcroft FM, Rorsman P (2013) K (ATP) channels and islet hormone secretion: new insights and controversies. Nat Rev Endocrinol 9:660–669

    Article  CAS  Google Scholar 

  • Atkinson MA, Eisenbarth GS, Michels AW (2014) Type 1 diabetes. Lancet 383(9911):69–82

    Article  Google Scholar 

  • Atta MS, Almadaly EA, Ali HE, Saleh RM, Assar DH, Al Jaouni SK, Mousa SA (2017) Thymoquinone defeats diabetes-induced testicular damage in rats targeting antioxidant, inflammatory and aromatase expression. Int J Mol Sci 18:919

    Article  CAS  Google Scholar 

  • Bajracharya GB (2015) Diversity, pharmacology and synthesis of bergenin and its derivatives: potential materials for therapeutic usages. Fitoterapia 101:133–152

    Article  CAS  Google Scholar 

  • Bancroft JD, Gamble MN (2002) Theory and practice of histological techniques, 5th edn. Churchill-Livingstone, Edinburgh, London, p 175

    Google Scholar 

  • Bioanalytical Method Validation (2013) US Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), Center for Veterinary Medicine (CVM): guidance for industry, bio-analytical method validation, Available online: http://www.fda.gov/downloads/drugs/guidancecomplianceregulatoryinformation/guidances/ucm368107.pdf

  • Cabral REL, Okada FK, Stumpp T, Vendramini V, Miraglia SM (2014) Carnitine partially protects the rat testis against the late damage produced by doxorubicin administered during pre-puberty. Andrology 2:931–942

    Article  CAS  Google Scholar 

  • Cheesbrough M (2009) District laboratory practice in tropical countries, 2nd edn. Vol. Part 1. Cambridge University Press, Cambridge, pp 1–454

    Google Scholar 

  • Ding GL, LiuY LME, Pan JX, Guo MX, Sheng JZ, Huang HF (2015) The effects of diabetes on male fertility and epigenetic regulation during spermatogenesis. Asian J Androl 17:948–953

    Article  CAS  Google Scholar 

  • Dkhil MA, Zrieq R, Al-Quraishy S, Abdel Moneim AE (2016) Selenium nanoparticles attenuate oxidative stress and testicular damage in streptozotocin-induced diabetic rats. Molecules 21(11):1517

    Article  CAS  Google Scholar 

  • European Medicines Agency (2011) Guideline on quality of herbal medicinal products/traditional herbal medicinal products: EMA/CPMP/QWP/2819/00 Rev.2. pp1–13 http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2011/09/WC500113209.pdf

  • Faid I, Al-Hussaini H, Kilarkaje N (2015) Resveratrol alleviates diabetes-induced testicular dysfunction by inhibiting oxidative stress and c-Jun N-terminal kinase signaling in rats. Toxicol Appl Pharmacol 289(3):482–494

    Article  CAS  Google Scholar 

  • Filler R (1993) Methods for evaluation of rat epididymal sperm morphology. In: Chapin RE, Heindel JJ (eds) Methods in toxicology: Part A: male reproductive toxicology. Academic Press, San Diego, CA, pp 335–343

    Google Scholar 

  • Frandsen CS, Dejgaard TF, Madsbad S (2016) Non-insulin drugs to treat hyperglycaemia in type 1 diabetes mellitus. Lancet Diabetes Endocrinol 4(9):766–780

    Article  CAS  Google Scholar 

  • Gurusubramanian G, Roy VK (2014) Expression of visfatin in alloxan-induced diabetic rat testis. Acta Histochem 116:1462–1468

    Article  CAS  Google Scholar 

  • Habig WH, Pabst MJ, Jakoby WB (1974) The first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139

    CAS  Google Scholar 

  • Hammer O, Harper DAT, Ryan PD (2001) PAST: palaeontological statistics software package for education and data analysis (version 1.86b). Palaeontol Electron 4:1–9

    Google Scholar 

  • Han X, Tao YL, Deng YP, Yu JW, Cai J, Ren GF, Sun YN, Jiang GJ (2017) Metformin ameliorates insulitis in STZ-induced diabetic mice. Peer J 5:e3155

    Article  CAS  Google Scholar 

  • Hwang SH, Kim JH, Wang Z, Lee JY, Lim SS (2017) Analytical method for the validation of three polyphenols as a marker compound for the standardization of Solidago virgaurea sub sp. gigantean extracts and anti adipogenesis of harvesting time and location. J Anal Methods Chem 2017:1–9

    Article  CAS  Google Scholar 

  • Jangir RN, Jain GC (2014) Diabetes mellitus induced impairment of male reproductive functions: a review. Curr Diabetes Rev 10:147–157

    Article  CAS  Google Scholar 

  • Jeremy M, Gurusubramanian G, Roy VK (2017) Localization pattern of visfatin (NAMPT) in D-galactose induced aged rat testis. Ann Anat 211:46–54

    Article  Google Scholar 

  • Johnsen SG (1970) Testicular biopsy score count - a method for registration of spermatogenesis in human testes. Normal values and results of 335 hypogonadal males. Hormones 1:2–25

    CAS  Google Scholar 

  • Kanter M, Aktas C, Erboga M (2013) Curcumin attenuates testicular damage, apoptotic germ cell death, and oxidative stress in streptozotocin-induced diabetic rats. Mol Nutr Food Res 57:1578–1585

    Article  CAS  Google Scholar 

  • Khan MH, Yadava PS (2010) Anti-diabetic plants used in Thoubal district of Manipur, Northeast India. Indian J Tradit Know 9:510–514

    Google Scholar 

  • Khan H, Amin H, Ullah A, Saba S, Rafique J, Khan K, Ahmad N, Badshah SL (2016) Antioxidant and anti plasmodial activities of bergenin and 11-O-galloylbergenin isolated from Mallotus philippensis. Oxid Med Cell Longev 2016:1051925

    Article  CAS  Google Scholar 

  • Khushboo M, Murthy MK, Devi MS, Sanjeev S, Ibrahim KS, Kumar NS, Roy VK, Gurusubramanian G (2018) Testicular toxicity and sperm quality following copper exposure in Wistar albino rats: ameliorative potentials of L-carnitine. Environ Sci Pollut Res Int 25(2):1837–1862

    Article  CAS  Google Scholar 

  • Kianifard D, Sadrkhanlou RA, Hasanzadeh S (2012) The ultrastructural changes of the Sertoli and Leydig cells following streptozotocin induced diabetes. Iran J Basic Med Sci 15(1):623–635

    Google Scholar 

  • Kilarkaje N, Al-Hussaini H, Al-Bader MM (2014) Diabetes-induced DNA damage and apoptosis are associated with poly (ADP ribose) polymerase 1 inhibition in the rat testis. Eur J Pharmacol 737:29–40

    Article  CAS  Google Scholar 

  • Kumar R, Patel DK, Prasad SK, Laloo D, Krishnamurthy S, Hemalatha S (2012) Type 2 anti-diabetic activity of bergenin from the roots of Caesalpinia digyna Rottler. Fitoterapia 83:395–401

    Article  CAS  Google Scholar 

  • La Vignera S, Condorelli R, Vicari E, D’Agata R, Calogero AE (2012) Diabetes mellitus and sperm parameters. J Androl 33:145–153

    Article  CAS  Google Scholar 

  • Lamprianou S, Gysemans C, Bou Saab J, Pontes H, Mathieu C, Meda P (2016) Glibenclamide Prevents Diabetes in NOD Mice. PLoS ONE 11(12):e0168839

    Article  CAS  Google Scholar 

  • Li BH, Wu JD, Li XL (2013) LC–MS/MS determination and pharmacokinetic study of bergenin, the main bioactive component of Bergenia purpurascens after oral administration in rats. J Pharm Anal 3(4):229–234

    Article  CAS  Google Scholar 

  • Lim HK, Kim HS, Choi HS, Choi J, Kim SH, Chang MJ (2001) Effects of bergenin, the major constituent of Mallotus japonicus against D-galactosamine-induced hepatotoxicity in rats. Pharmacology 63(2):71–75

    Article  CAS  Google Scholar 

  • Long L, Wang J, Lu X, Xu Y, Zheng S, Luo C, Li Y (2015) Protective effects of scutellarin on type II diabetes mellitus-induced testicular damages related to reactive oxygen species/Bcl-2/Bax and reactive oxygen species/microcirculation/staving pathway in diabetic rat. J Diabetes Res 2015:252530

    Article  CAS  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    CAS  Google Scholar 

  • Maresch CC, Stute DC, Ludlow H, Hammes HP, de Kretser DM, Hedger MP, Linn T (2017) Hyperglycemia is associated with reduced testicular function and activin dysregulation in the Ins2Akita+/- mouse model of type 1 diabetes. Mol Cell Endocrinol 446:91–101

    Article  CAS  Google Scholar 

  • Meistrich ML, van Beek MEAB (1993) Spermatogonial stem cells: assessing their survival and ability to produce differentiated cells. In: Chapin RE, Heindel J (eds) Methods in toxicology, vol 3A Academic Press, New York, pp 106-123

  • National Pharmacopoeia Committee (2015) Pharmacopoeia of People’s Republic of China, Part 2, China Medical. Science Press, Beijing

    Google Scholar 

  • Navarro-Casado L, Juncos-Tobarra MA, Chafer-Rudilla M, de Onzono LI, Blazquez-Cabrera JA, Miralles-Garcia JM (2010) Effect of experimental diabetes and STZ on male fertility capacity study in rats. J Androl 31:584–592

    Article  CAS  Google Scholar 

  • Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358

    Article  CAS  Google Scholar 

  • Omar SS, Aly RG, Badae NM (2018) Vitamin E improves testicular damage in streptozocin‐induced diabetic rats, via increasing vascular endothelial growth factor and poly (ADP-ribose) polymerase-1. Andrologia 50(3):e12925

    Article  CAS  Google Scholar 

  • Organisation for Economic Co-operation and Development (OECD) (2001) Guideline for the Testing of Chemicals. In: Acute oral toxicity-fixed dose procedure, vol 420. OECD/OCDE, Paris, France, pp 1–14

    Google Scholar 

  • Pitetti JL, Calvel P, Zimmermann C, Conne B, Papaioannou MD, Aubry F, Cederroth CR, Urner F, Fumel B, Crausaz M, Docquier M, Herrera PL, Pralong F, Germond M, Guillou F, Jégou B, Nef S (2013) An essential role for insulin and IGF1 receptors in regulating sertoli cell proliferation, testis size, and FSH action in mice. Mol Endocrinol 27(5):814–827

    Article  CAS  Google Scholar 

  • Rama Raju G, Jaya Prakash G, Murali Krishna K, Madan K, Siva T, Narayana C, Ravi Krishna H (2012) Noninsulin-dependent diabetes mellitus: effects on sperm morphological and functional characteristics, nuclear DNA integrity and outcome of assisted reproductive technique. Andrologia 44:490–498

    Article  CAS  Google Scholar 

  • Rashid K, Sil PC (2015) Curcumin ameliorates testicular damage in diabetic rats by suppressing cellular stress-mediated mitochondria and endoplasmic reticulum-dependent apoptotic death. Biophys Acta 1852:70–82

    CAS  Google Scholar 

  • Ren X, Ma S, Wang J, Tian S, Fu X, Liu X, Li Z, Zhao B, Wang X (2016) Comparative effects of dexamethasone and bergenin on chronic bronchitis and their anti-inflammatory mechanisms based on NMR metabolomics. Mol Biosyst 12(6):1938–1947

    Article  CAS  Google Scholar 

  • Ricci G, Catizone A, Esposito R, Pisanti FA, Vietri MT, Galdieri M (2009) Diabetic rat testes: morphological and functional alterations. Andrologia 41:361–368

    Article  CAS  Google Scholar 

  • Robb GW, Amann PR, Killian GJ (1978) Daily sperm production and epididymal sperm reserves of pubertal and adult rats. J Reprod Fertil 54:103–107

    Article  CAS  Google Scholar 

  • Roy VK, Chenkual L, Gurusubramanian G (2015) Protection of testis through antioxidant action of Mallotus roxburghianus in alloxan-induced diabetic rat model. J Ethnopharmacol 176:268–280

    Article  CAS  Google Scholar 

  • Roy VK, Peki V, Devi MS, Sanjeev S, Khusboo M, Zothansanga R, Ibrahim KS, Kumar NS, Gurusubramanian G (2017) Biosterilant effects of Bacillus thuringiensis kurstaki HD-73 extract on male Wistar albino rats. Theriogenology 88:73–83

    Article  Google Scholar 

  • Sergerie M, Laforest G, Bujan L, Bissonnette F (2005) Sperm DNA fragmentation: threshold value in male fertility. Hum Reprod 20:3446–3451

    Article  CAS  Google Scholar 

  • Subramanian R, Subbramaniyan P, Raj V (2015) Isolation of bergenin from Peltophorum pterocarpum flowers and its bioactivity. Beni-Seuf Univ. J Appl Sci 42:256–261

    Google Scholar 

  • Sumino M, Sekine T, Ruangrungsi N, Ikegami F (2002) Ardisiphenols and other antioxidant principles from the fruits of Ardisia colorata. Chem Pharm Bull 50:1484–1487

    Article  CAS  Google Scholar 

  • World Health Organization (2013) WHO traditional medicine strategy 2014–2023. World Health Organization, WHO Press, Geneva, Switzerland

    Google Scholar 

Download references

Acknowledgements

The authors acknowledge the instrumentation facility in Mizoram University funded by Department of Biotechnology, Government of India, New Delhi - Bioinformatics Infrastructure Facility (No. BT/BI/12/060/2012(NERBIF-MUA) and State Biotech Hub Programme (No. BT/04/NE/2009).

Author information

Authors and Affiliations

  1. Department of Zoology, Mizoram Central University, Aizawl, Mizoram, 796004, India

    Sanasam Sanjeev, Meesala Krishna Murthy, Maibam Sunita Devi, Maurya Khushboo, Zothanmawii Renthlei, Vikas Kumar Roy & Guruswami Gurusubramanian

  2. Department of Biotechnology, Mizoram Central University, Aizawl, Mizoram, 796004, India

    Kalibulla Syed Ibrahim & Nachimuthu Senthil Kumar

Authors
  1. Sanasam Sanjeev
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Meesala Krishna Murthy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maibam Sunita Devi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Maurya Khushboo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zothanmawii Renthlei
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kalibulla Syed Ibrahim
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Nachimuthu Senthil Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Vikas Kumar Roy
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Guruswami Gurusubramanian
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guruswami Gurusubramanian.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Responsible editor: Philippe Garrigues

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

ESM 1

(PDF 1726 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sanjeev, S., Murthy, M.K., Sunita Devi, M. et al. Isolation, characterization, and therapeutic activity of bergenin from marlberry (Ardisia colorata Roxb.) leaf on diabetic testicular complications in Wistar albino rats. Environ Sci Pollut Res 26, 7082–7101 (2019). https://doi.org/10.1007/s11356-019-04139-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11356-019-04139-9

Keywords

Search

Navigation