Study on the Antileukemic Immunity of Dunalella salina and Hepatoprotective Activity of Hydrogen-Rich Water in Mice

• Main Authors: Wen-Chen Chuang, 莊文貞
• Other Authors: Jiunn-Wang Liao
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/q88h28

博士 === 國立中興大學 === 獸醫學系暨研究所 === 103 === part 1 Dunaliella salina (D. salina) has been shown to have antioxidant property and induce apoptotic cell death of human cancer cell in vitro. However, there is no information available on anti-leukemia effect or immunomodulatory activity in vivo. In this study, we applied D. salina to syngeneic leukemia-implanted mice (BALB/c and WEHI-3) to investigate its immunological and anti-leukemia properties. Oral administration of D. salina (184.5, 369 and 922.5 mg/kg) for 4 weeks could inhibit tumor metastasis into spleen and liver, and prolonged the survival rate in BALB/c leukemia mice, they were received an intravenous injection of WEHI-3 cells. It had also increased the population and proliferation of T cells (CD3) and B cells (CD19) in splenocytes following Con A/LPS stimulation. Furthermore, D. salina increased the phagocytosis of macrophages and enhanced the cytotoxicity of natural killer cells on flow cytometry assay. Moreover, D. salina enhanced the levels of interferon-γ and interleukin 2 (IL-2) but reduced the levels of IL-4 and IL-10 in leukemia mice. In conclusion, these results demonstrated that the application of D. salina had beneficial effects to WEHI-3 leukemic mice by prolonging survival via modulating the immune responses. part 2 Hydrogen, a new antioxidant, was reported to selectively reduce the strongest oxidants and alleviates acute oxidative stress in many models. The protective effects of hydrogen-rich water (HRW) against liver damage were evaluated by its attenuation of carbon tetrachloride (CCl4)-induced hepatotoxicity in mice. Male ICR mice were randomly divided into control, CCl4, CCl4 + Silymarin, CCl4 + HPW, CCl4 + HRW, and CCl4 + Silymarin +HRW groups. Treatment with CCl4 for 8 weeks could induce liver lesions included leukocytes infiltration, hepatocyte vacuolization, necrosis, fibrosis and an increase of serum alanine aminotransferase (ALT), and aminotransferase (AST) activity. In addition, CCl4 significantly increased lipid oxidation, decreased the activities of superoxide dismutase (SOD), catalase, glutathione reductase (GSH-Rd) and glutathione peroxidase (GSH-Px), and increased the inflammatory cytokines of TNF-α and IL-6 in the liver. By contrast, Silymarin plus HRW supplement significantly ameliorated the CCl4-induced liver lesions, lowered the serum levels of hepatic enzyme markers (ALT and AST), increased the activities of antioxidant enzyme and reduced the lipid peroxidation and inflammatory markers of TNF-α and IL-6 in liver compared to Silymarin treatment. Consistent with these observations, the proteins expression of iNOS, COX-2, HMGB1, HSP70, HIF-1α inhibited by Silymarin plus HRW treatment. Silymarin plus HRW also caused a significant increase in antioxidant proteins including HO-1, γ-GCS. Moreover, significant decreases of MMP-9, MMP-2, phosphorylated ERK (p-ERK), AKT (p- AKT), JNK (p-JNK) and NF-κB (p-P65) reduced by sylimarin plus HRW treatment as compared to Silymarin treatment. Therefore, the results of this study reveal that HRW can be proposed to enhance the protecting effects by Silymarin on CCl4-induced inflammation and oxidative stress of liver in mice, and the hepatoprotective effect might be correlated with its free radical scavenging effect on enhancing the activity of antioxidant-defense system and inhibiting of inflammatory response.