Jun Zhou1, Qin Wang, Qunwei Wang, Wenbin Duan. 1. Department of Minimal Invasive Surgery, Second Xiangya Hospital, Central South University, Changsha 410011, China.
Abstract
OBJECTIVE: To study the effects of norcantharidin (NCTD) on lipopolysaccharide (LPS)-induced hepatocyte injury and the expression of TNF-α and IL-6 in vitro. METHODS: Hepatocytes were isolated from male Sprague-Dawley rats by collagenase perfusion. LPS at concentration of 40 mg/L was used to induce injury to the cultured cells, and NCTD (0.5, 1.0, 2.5 μg/mL) was added at the same time. After 24 h of incubation, the cell proliferation rates were detected by MTT. LDH, TNF-α and IL-6 were measured by appropriate reagent kits.NF-κB DNA binding activity was measured. RESULTS: 40 mg/L LPS caused a 27% growth inhibition in primary hepatocytes. LDH leakage was 20- fold higher in NCTD-treated hepatocytes than in normal ones. TNF-α and IL-6 expression significantly increased. In cells treated with NCTD at doses of 0.5, 1.0 and 2.5 μg/mL, LDH leakage, TNF-α and IL-6 expression, and NF-κB DNA binding activity were attenuated in a dose dependent manner. CONCLUSION: NCTD protects hepatocytes from injury induced by LPS; the protection is associated with suppression of the inflammatory cytokine TNF-α and IL-6.
OBJECTIVE: To study the effects of norcantharidin (NCTD) on lipopolysaccharide (LPS)-induced hepatocyte injury and the expression of TNF-α and IL-6 in vitro. METHODS: Hepatocytes were isolated from male Sprague-Dawley rats by collagenase perfusion. LPS at concentration of 40 mg/L was used to induce injury to the cultured cells, and NCTD (0.5, 1.0, 2.5 μg/mL) was added at the same time. After 24 h of incubation, the cell proliferation rates were detected by MTT. LDH, TNF-α and IL-6 were measured by appropriate reagent kits.NF-κB DNA binding activity was measured. RESULTS: 40 mg/L LPS caused a 27% growth inhibition in primary hepatocytes. LDH leakage was 20- fold higher in NCTD-treated hepatocytes than in normal ones. TNF-α and IL-6 expression significantly increased. In cells treated with NCTD at doses of 0.5, 1.0 and 2.5 μg/mL, LDH leakage, TNF-α and IL-6 expression, and NF-κB DNA binding activity were attenuated in a dose dependent manner. CONCLUSION: NCTD protects hepatocytes from injury induced by LPS; the protection is associated with suppression of the inflammatory cytokine TNF-α and IL-6.