Metabolic Brain Disease

, Volume 33, Issue 5, pp 1617–1624 | Cite as

Methylenetetrahydrofolate reductase C677T variant and hyperhomocysteinemia in subarachnoid hemorrhage patients from India

  • Mohit Kumar
  • Sachin Goudihalli
  • Kanchan Mukherjee
  • Sivashanmugam Dhandapani
  • Rajat SandhirEmail author
Original Article


Methylenetetrahydrofolate reductase (MTHFR) polymorphism (C677T, A1298C) has been implicated in increased plasma homocysteine (Hcy) levels. The present study was designed to investigate the association between MTHFR polymorphism and increased Hcy levels in subarachnoid haemorrhage (SAH) patients. A total of 150 subjects from North India were included in the study, comprising of 100 SAH patients and 50 healthy controls. Plasma Hcy levels was determined and MTHFR polymorphism (C677T, A1298C) was screened by High resolution melting (HRM) analysis. Plasma Hcy levels were found to be significantly higher (p < 0.001) in SAH patients than in healthy controls. No significant difference in the genotype and allele frequency of MTHFR A1298C was observed. However, frequency of MTHFR C677T genotype, CT (53% vs. 20%; p < 0.001) and TT (15% vs. 2%; p < 0.05) was significantly higher in SAH group as compared to healthy controls. The frequency of T allele (41.5% vs. 12%; p < 0.001) was also found to be higher in SAH patients in comparison to healthy controls. Furthermore, Hcy levels were higher in SAH patients with TT genotype than in patients having CT genotype, whereas CC genotype had lower Hcy levels. The study suggests that higher frequency of MTHFR C677T allele may contribute to etiopathology of SAH through increase in Hcy levels.


Homocysteine Methylenetetrahydrofolate reductase MTHFR A1298C MTHFR C677T Subarachnoid haemorrhage Single nucleotide polymorphism 



Coronary artery diseases


High resolution melting






Methylenetetrahydrofolate reductase


Single nucleotide polymorphism


Subarachnoid haemorrhage


7-benzo-2-oxa-1,3-diazole-4-sulfonic acid





The financial assistance provided by Department of Bio-Technology (DBT), Govt. of India, New Delhi [grant number BT/361/NE/TBP/2012] is acknowledged. The fellowship to MK from the University Grants Commission (UGC), New Delhi [F.17-7(J)/2004(SA-1)] is also acknowledged. The authors also acknowledge UGC-Special Assistance Programme (UGC-SAP) Departmental Research Support-II (DRS-II). The financial assistance provided by the Department of Science and Technology (DST), New Delhi under Promotion of University Research and Scientific Excellence (PURSE) grant is also acknowledged. The help of Prof. Suresh K. Sharma from the Department of Statistics, Panjab University in data analysis is gratefully acknowledged.

Compliance with ethical standards

Ethical approval

All experiments were accordance with the ethical standards and were approved by the Institutional Ethics Committee of PGIMER (NK/872/M.Ch./542).

Informed consent

Informed consent was obtained from all participants included in the study.

Conflict of interest

The authors state no conflict of interest.


  1. Aggarwal A, Dhandapani S, Praneeth K et al (2018) Comparative evaluation of H&H and WFNS grading scales with modified H&H (sans systemic disease): A study on 1000 patients with subarachnoid hemorrhage. Neurosurg Rev 41:241–247. CrossRefPubMedGoogle Scholar
  2. Al-Allawi N, Avo A, Jubrael J (2009) Methylenetetrahydrofolate reductase C677T polymorphism in Iraqi patients with ischemic stroke. Neurol India 57:631. CrossRefPubMedGoogle Scholar
  3. Alluri RV, Mohan V, Komandur S et al (2005) MTHFR C677T gene mutation as a risk factor for arterial stroke: a hospital based study. Eur J Neurol 12:40–44. CrossRefPubMedGoogle Scholar
  4. Amouzou EK, Chabi NW, Adjalla CE et al (2004) High prevalence of hyperhomocysteinemia related to folate deficiency and the 677C>T mutation of the gene encoding methylenetetrahydrofolate reductase in coastal West Africa. Am J Clin Nutr 79:619–624CrossRefPubMedGoogle Scholar
  5. Angeline T, Jeyaraj N, Granito S, Tsongalis GJ (2004) Prevalence of MTHFR gene polymorphisms (C677T and A1298C) among Tamilians. Exp Mol Pathol 77:85–88. CrossRefPubMedGoogle Scholar
  6. Banerjee I, Gupta V, Ganesh S (2007) Association of gene polymorphism with genetic susceptibility to stroke in Asian populations: a meta-analysis. J Hum Genet 52:205–219. CrossRefPubMedGoogle Scholar
  7. Bhagwati SN (1998) Incidence of subarachnoid hemorrhage from aneurysmal rupture in India. Neurol Med Chir 38(Suppl):128–130CrossRefGoogle Scholar
  8. Bhargava S, Parakh R, Manocha A et al (2007) Prevalence of Hyperhomocysteinemia in Vascular Disease: Comparative Study of Thrombotic Venous Disease Vis-è-Vis Occlusive Arterial Disease. Vascular 15:149–153. CrossRefPubMedGoogle Scholar
  9. Bhat AR, Afzalwani M, Kirmani AR (2011) Subarachnoid hemorrhage in Kashmir: Causes, risk factors, and outcome. Asian J Neurosurg 6:57–71. CrossRefPubMedPubMedCentralGoogle Scholar
  10. Biswas A, Ranjan R, Meena A et al (2009) Homocystine Levels, Polymorphisms and the Risk of Ischemic Stroke in Young Asian Indians. J Stroke Cerebrovasc Dis 18:103–110. CrossRefPubMedGoogle Scholar
  11. Cacciapuoti F (2013) Lowering homocysteine levels with folic acid and B-vitamins do not reduce early atherosclerosis, but could interfere with cognitive decline and Alzheimer’s disease. J Thromb Thrombolysis 36:258–262. CrossRefPubMedGoogle Scholar
  12. Cronin S, Furie KL, Kelly PJ (2005) Dose-Related Association of MTHFR 677T Allele With Risk of Ischemic Stroke: Evidence From a Cumulative Meta-Analysis. Stroke 36:1581–1587. CrossRefPubMedGoogle Scholar
  13. Dayakar S, Goud KI, Reddy TPK et al (2011) Sequence Variation of the Methylene Tetrahydrofolate Reductase Gene (677C>T and 1298A>C) and Traditional Risk Factors in a South Indian Population. Gen Test Mol Biomarkers 15:765–769. CrossRefPubMedGoogle Scholar
  14. Dhandapani SS, Manju D, Vivekanandhan S et al (2010) Prospective longitudinal study of biochemical changes in critically ill patients with severe traumatic brain injury: Factors associated and outcome at 6 months. Indian J Neurotrauma 7:23–27. CrossRefGoogle Scholar
  15. Dhandapani S, Aggarwal A, Srinivasan A et al (2015a) Serum lipid profile spectrum and delayed cerebral ischemia following subarachnoid hemorrhage: Is there a relation?. Surgical neurology international. 2015 6(Suppl 21):S543. CrossRefGoogle Scholar
  16. Dhandapani S, Goudihalli S, Mukherjee KK et al (2015b) Prospective study of the correlation between admission plasma homocysteine levels and neurological outcome following subarachnoid hemorrhage: A case for the reverse epidemiology paradox? Acta Neurochir 157:399–407. CrossRefPubMedGoogle Scholar
  17. Dhandapani S, Bajaj A, Gendle C et al (2018) Independent impact of plasma homocysteine levels on neurological outcome following head injury. Neurosurg Rev 41:513–517. CrossRefPubMedGoogle Scholar
  18. Duan W, Ladenheim B, Cutler RG et al (2002) Dietary folate deficiency and elevated homocysteine levels endanger dopaminergic neurons in models of Parkinson’s disease. J Neurochem 80:101–110CrossRefPubMedGoogle Scholar
  19. Ganguly P, Alam SF (2015) Role of homocysteine in the development of cardiovascular disease. Nutr J 14:6. CrossRefPubMedPubMedCentralGoogle Scholar
  20. Hofmann MA, Lalla E, Lu Y et al (2001) Hyperhomocysteinemia enhances vascular inflammation and accelerates atherosclerosis in a murine model. J Clin Invest 107:675–683. CrossRefPubMedPubMedCentralGoogle Scholar
  21. Hoque MM, Rahman MZ, Rahman MR (2008) Role of homocysteine in cerebrovascular disease. Mymensingh Medical Journal : MMJ 17:S39–S42PubMedGoogle Scholar
  22. Jiang S, Li J, Zhang Y et al (2017) Methylenetetrahydrofolate reductase C677T polymorphism, hypertension and risk of stroke: a prospective, nested case-control study. Int J Neurosci 127:253–260. CrossRefPubMedGoogle Scholar
  23. Kalita J, Srivastava R, Bansal V et al (2006) Methylenetetrahydrofolate reductase gene polymorphism in Indian stroke patients. Neurol India 54:260–263. CrossRefPubMedGoogle Scholar
  24. Kang SS, Wong PW, Zhou JM et al (1988) Thermolabile methylenetetrahydrofolate reductase in patients with coronary artery disease. Metab Clin Exp 37:611–613CrossRefPubMedGoogle Scholar
  25. Kang S, Zhao X, Liu L et al (2013) Association of the C677T polymorphism in the MTHFR gene with hemorrhagic stroke: a meta-analysis. Gen Test Mol Biomarkers 17:412–417. CrossRefGoogle Scholar
  26. Kumar J, Das SK, Sharma P et al (2005) Homocysteine levels are associated with MTHFR A1298C polymorphism in Indian population. J Hum Genet 50:655–663. CrossRefPubMedGoogle Scholar
  27. Kumar M, Modi M, Sandhir R (2017) Hydrogen sulfide attenuates homocysteine-induced cognitive deficits and neurochemical alterations by improving endogenous hydrogen sulfide levels. BioFactors 43:434–450. CrossRefPubMedGoogle Scholar
  28. Li Z, Sun L, Zhang H et al (2003) Elevated Plasma Homocysteine Was Associated With Hemorrhagic and Ischemic Stroke, but Methylenetetrahydrofolate Reductase Gene C677T Polymorphism Was a Risk Factor for Thrombotic Stroke: A Multicenter Case-Control Study in China. Stroke 34:2085–2090. CrossRefPubMedGoogle Scholar
  29. Liu S, Wu Y, Liu X et al (2017) Lack of association between MTHFR A1298C variant and Alzheimer’s disease: evidence from a systematic review and cumulative meta-analysis. Neurol Res 39:426–434. CrossRefPubMedGoogle Scholar
  30. Lv Q-Q, Lu J, Sun H, Zhang J-S (2015) Association of methylenetetrahydrofolate reductase (MTHFR) gene polymorphism with ischemic stroke in the Eastern Chinese Han population. Genet Mol Res 14:4161–4168. CrossRefPubMedGoogle Scholar
  31. Mansoori N, Tripathi M, Luthra K, et al (2012) MTHFR (677 and 1298) and IL-6-174 G/C genes in pathogenesis of Alzheimer’s and vascular dementia and their epistatic interaction. Neurobiol Aging 33:1003.e1–1001003.e8. CrossRefGoogle Scholar
  32. Modi M, Prabhakar S, Majumdar S et al (2005) Hyperhomocysteinemia as a risk factor for ischemic stroke: an Indian scenario. Neurol India 53:297-301-2Google Scholar
  33. Moe KT, Woon FP, De Silva DA et al (2008) Association of acute ischemic stroke with the MTHFR C677T polymorphism but not with NOS3 gene polymorphisms in a Singapore population. Eur J Neurol 15:1309–1314. CrossRefPubMedGoogle Scholar
  34. Morita H, Kurihara H, Tsubaki S et al (1998) Methylenetetrahydrofolate reductase gene polymorphism and ischemic stroke in Japanese. Arterioscler Thromb Vasc Biol 18:1465–1469CrossRefPubMedGoogle Scholar
  35. Najafipour R, Moghbelinejad S, Aleyasin A, Jalilvand A (2017) Effect of B9 and B12 vitamin intake on semen parameters and fertility of men with MTHFR polymorphisms. Andrology 5:704–710. CrossRefPubMedGoogle Scholar
  36. Norambuena PA, Copeland JA, Křenková P et al (2009) Diagnostic method validation: High resolution melting (HRM) of small amplicons genotyping for the most common variants in the MTHFR gene. Clin Biochem 42:1308–1316. CrossRefPubMedGoogle Scholar
  37. Panigrahi I, Chatterjee T, Biswas A et al (2006) Role of MTHFR C677T polymorphism in ischemic stroke. Neurol India 54:48-50-2Google Scholar
  38. Rai V (2016a) Folate Pathway Gene Methylenetetrahydrofolate Reductase C677T Polymorphism and Alzheimer Disease Risk in Asian Population. Indian J Clin Biochem 31:245–252. CrossRefPubMedGoogle Scholar
  39. Rai V (2016b) Association of methylenetetrahydrofolate reductase (MTHFR) gene C677T polymorphism with autism: evidence of genetic susceptibility. Metab Brain Dis 31:727–735. CrossRefPubMedGoogle Scholar
  40. Roussotte FF, Hua X, Narr KL et al (2017) Archival Report The C677T Variant in MTHFR Modulates Associations Between Brain Integrity, Mood, and Cognitive Functioning in Old Age. Biol Psychiatry. Google Scholar
  41. Sambrook J, Russell DW (2006) Purification of nucleic acids by extraction with phenol:chloroform. CSH Protocols 2006. CrossRefPubMedGoogle Scholar
  42. Saraswathy KN, Mukhopadhyay R, Sinha E et al (2008) MTHFR C677T polymorphisms among the Ahirs and Jats of Haryana (India). Am J Hum Biol 20:116–117. CrossRefPubMedGoogle Scholar
  43. Sawuła W, Banecka-Majkutewicz Z, Kadziński L et al (2008) Improved HPLC method for total plasma homocysteine detection and quantification. Acta Biochim Pol 55:119–125PubMedGoogle Scholar
  44. Serapinas D, Boreikaite E, Bartkeviciute A et al (2017) The importance of folate, vitamins B6 and B12 for the lowering of homocysteine concentrations for patients with recurrent pregnancy loss and MTHFR mutations. Reprod Toxicol 72:159–163. CrossRefPubMedGoogle Scholar
  45. Škovierová H, Vidomanová E, Mahmood S, et al (2016) The Molecular and Cellular Effect of Homocysteine Metabolism Imbalance on Human Health. Int J Mol Sci 17. CrossRefPubMedCentralGoogle Scholar
  46. Smith AD, Smith SM, de Jager CA et al (2010) Homocysteine-lowering by b vitamins slows the rate of accelerated brain atrophy in mild cognitive impairment: A randomized controlled trial. PLoS One 5:1–10. CrossRefGoogle Scholar
  47. Tripathi R, Tewari S, Singh PK, Agarwal S (2010) Association of homocysteine and methylene tetrahydrofolate reductase (MTHFR C677T) gene polymorphism with coronary artery disease (CAD) in the population of North India. Genet Mol Biol 33:224–228. CrossRefPubMedPubMedCentralGoogle Scholar
  48. van Gijn J, Kerr RS, Rinkel GJE et al (2007) Subarachnoid haemorrhage. Lancet (London, England) 369:306–318. CrossRefGoogle Scholar
  49. Wilcken B, Bamforth F, Li Z et al (2003) Geographical and ethnic variation of the 677C>T allele of 5,10 methylenetetrahydrofolate reductase (MTHFR): findings from over 7000 newborns from 16 areas world wide. J Med Genet 40:619–625. CrossRefPubMedPubMedCentralGoogle Scholar
  50. Yadav U, Kumar P, Gupta S, Rai V (2016) Role of MTHFR C677T gene polymorphism in the susceptibility of schizophrenia: An updated meta-analysis. Asian J Psychiatr 20:41–51. CrossRefPubMedGoogle Scholar
  51. Yao E-S, Tang Y, Xie M-J et al (2016) Elevated Homocysteine Level Related to Poor Outcome After Thrombolysis in Acute Ischemic Stroke. Med Sci Monit 22:3268–3273CrossRefPubMedPubMedCentralGoogle Scholar
  52. Zhao M, Wang X, He M et al (2017) Homocysteine and Stroke Risk. Stroke 48:1183–1190. CrossRefPubMedGoogle Scholar
  53. Zhou F, chen B, Chen C et al (2015) Elevated Homocysteine Levels Contribute to Larger Hematoma Volume in Patients with Intracerebral Hemorrhage. J Stroke Cerebrovasc Dis 24:784–788. CrossRefPubMedGoogle Scholar
  54. Zoccolella S, dell’Aquila C, Specchio LM et al (2010) Elevated homocysteine levels in Parkinson’s Disease: is there anything besides L-dopa treatment? Curr Med Chem 17:213–221CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Mohit Kumar
    • 1
  • Sachin Goudihalli
    • 2
  • Kanchan Mukherjee
    • 2
  • Sivashanmugam Dhandapani
    • 2
  • Rajat Sandhir
    • 1
    Email author
  1. 1.Department of Biochemistry, Basic Medical Sciences Block-IIPanjab UniversityChandigarhIndia
  2. 2.Department of NeurosurgeryPost Graduate Institute of Medical Education and ResearchChandigarhIndia

Personalised recommendations