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Cover page of the Journal of Health Sciences
Year : 2018  |  Volume : 11  |  Issue : 1  |  Page : 12-18

Vitamin B12 insufficiency in excess folic acid downregulates methylenetetrahydrofolate reductase gene and increases homocysteine, tumor necrosis factor-alpha, and oxidative stress in hepatocytes

1 Dr. Prabhakar Kore Basic Science Research Centre, KLE Academy of Higher Education and Research, KLE University, Belagavi, Karnataka, India
2 Department of Biotechnology, Sinhgad College of Engineering, SP Pune University, Vadgaon Budruk, Pune, Maharashtra, India

Correspondence Address:
Dr. Sanjay Mishra
Dr. Prabhakar Kore Basic Science Research Centre, KLE University, Nehru Nagar, Belagavi - 590 010, Karnataka
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/kleuhsj.kleuhsj_128_17

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OBJECTIVE: Folate and Vitamin B12 (B12) are crucial for normal growth, development, and cellular functions. The aim of this study was to determine the effect of required and excess folic acid (FA) concentration and imbalance of FA and B12 caused due to folate fortification on hepatocytes health and function in vitro. MATERIALS AND METHODS: Human hepatocellular carcinoma (HepG2) cells were cultured in FA and B12 deficient media for 15 days and further were supplemented with required FA: (6 μM) and excess FA: (60 μM) individually and in combination with B12: (500 nM). We assessed HepG2 proliferation, viability, tumor necrosis factor-alpha (TNFα) and methylenetetrahydrofolate reductase (MTHFR) mRNA expression, homocysteine, and malondialdehyde levels. RESULTS: Supplementation of B12 with FA increased proliferation as compared to only FA supplemented group. FA and B12 deficient cells and excess FA supplementation resulted in decreased viability. Supplementation of B12 at both FA concentrations increased (P < 0.05) cell viability. In the presence of B12 oxidative stress, TNFα mRNA and TNFα levels in cell lysate significantly decreased (P < 0.05) in both FA supplemented cells as compared to excess FA cells. Homocysteine levels significantly decreased (P < 0.05) in cells supplemented with normal FA + B12 as compared to excess FA supplemented cells and FA and B12 deficient cells. Supplementation of excess FA significantly downregulated (P < 0.05) MTHFR mRNA levels as compared to required FA cells. Combination of FA + B12 significantly upregulated (P < 0.01) MTHFR mRNA levels as compared to excess FA. CONCLUSION: These results accord the promoting effect of B12 with FA in developing nations like India where B12 deficiency is common while the concentration of folate is adequate.

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