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Chronic hepatitis B virus (HBV) infection is a serious public health issue. Vitamin D is involved in various pathophysiological mechanisms as an immune modulator and the deficiency rate of vitamin D is prevalent in chronic liver disease. Fucoidan exerts anti-inflammatory, anticoagulant, antitumor, antimetastatic, and antiangiogenetic effects; however, its effect on the immune responses of HBV patients is unclear. This study investigated how 25(OH)Vitamin D status affected the effectiveness of oligo fucoidan in patients with HBV infection in the immune tolerance phase. Fifty-one patients received oligo fucoidan 4400 mg/day for 48 weeks. Flow cytometry was used to detect T lymphocyte markers (CD3⁺CD4⁺, CD3⁺CD8⁺, CD4⁺CD45RO⁺, CD8⁺CD45RO⁺). The levels of white blood cell (WBC), platelets (PLT), and albumin were decreased after 48 weeks of supplementation (<i>p &lt;</i> 0.05). Percentages of CD3⁺CD8⁺ and CD8⁺CD45RO+ cells were decreased after 12 weeks of supplementation (<i>p &lt;</i> 0.05). In patients with adequate vitamin D, HBV-DNA concentrations decreased and the proportion of CD4⁺CD45RO⁺ and CD8⁺CD45RO⁺ cells increased upon oligo fucoidan supplementation. The HBeAg status of one vitamin D-adequate patient changed from positive to negative at the 12th week of supplementation. The oligo fucoidan may regulate immune effects in patients with HBV infection, and the 25(OH)Vitamin D status might have affected the effectiveness of oligo fucoidan.