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KMID : 0615720170340010023
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Jouranl of Korean Acupuncture and Moxibustion society
2017 Volume.34 No. 1 p.23 ~ p.30
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Analysis of the Apoptotic Mechanisms of Snake Venom Toxin on Inflammation-induced HaCaT Cell-line
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Chun Youl-Woong
Song Ho-Sueb
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Abstract
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Objectives: In this study, the roles of Interleukin (IL)-4 and Signal transducer and activator of transcription 6 (STAT6), which have been reported to play a role in the pathogenesis of inflammation and cancer, were evaluated in snake venom toxin (SVT)-induced apoptosis.
Methods: Inflammation was induced in human HaCaT kerationocytes, by lipopolysaccharide (LPS; 1 μg/mL) or tumor necrosis factor-α (TNF-α), followed by treatment with SVT (0, 1, or 2 μg/mL). Cell viability was assessed by MTT assays after 24 h, and the expression of levels of IL-4, STAT6, and the apoptosis-related proteins p53, Bax, and Bcl-2 were evaluated by western blotting. Electro mobility shift assays (EMSAs) were performed to evaluate the DNA binding capacity of STAT6.
Results: MTT assays showed that inflammation-induced growth of HaCaT cells following LPS or TNF-α stimulation was inhibited by SVT. Western blot analysis showed that p53 and Bax, which promote apoptosis, were increased, whereas that of Bcl-2, an anti-apoptotic protein, was decreased in a concentration-dependent manner in LPS- or TNF-α-induced HaCaT cells following treatment with SVT. Moreover, following treatment of HaCaT cells with LPS, IL-4 concentrations were increased, and treatment with SVT further increased IL-4 expression in a concentration-dependent manner. Western blotting and EMSAs showed that the phosphorylated form of STAT6 was increased in HaCaT cells in the context of LPS- or TNF-α-induced inflammation in a concentration-dependent manner, concomitant with an increase in the DNA binding activity of STAT6.
Conclusion: SVT can effectively promote apoptosis in HaCaT cells in the presence of inflammation through a pathway involving IL-4 and STAT6.
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KEYWORD
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Snake venom toxin, Inflammation, Apoptosis, Interleukin-4, Signal transducer and activator of transcription 6
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