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目的:探讨新合成的硫化氢(H2S)供体GYY4137对小鼠原代肝脂肪变性细胞胞质内脂质分解的影响。方法:体外用油酸诱导肝细胞脂肪变性模型。采用两步原位灌流法分离C57BL/6小鼠原代肝细胞并分为4组:对照组用正常培养液培养54 h;模型组用含1.2 mmol/L油酸(溶于10%BSA)的培养液培养48 h,再用RPMI-1640培养液培养6 h;H2S组和DL-炔丙基甘氨酸(PAG;胱硫醚γ-裂解酶抑制剂,抑制H2S合成)组则用含有1.2mmol/L油酸的培养液培养48 h,再用无血清无酚红的RPMI-1640培养液(分别给予含有1 mmol/L GYY4137和200μmol/L PAG)培养6 h。测量细胞甘油释放量及胞内激素敏感性脂肪酶(HSL)的蛋白表达量。结果:和模型组相比,H_2S组培养液中甘油释放量和磷酸化HSL(p-HSL)蛋白表达水平明显降低,而PAG组又明显升高。结论:在小鼠原代脂肪变性肝细胞中,GYY4137可能通过抑制HSL的磷酸化水平而减少胞质内脂质分解。
Objective: To investigate the effect of newly synthesized hydrogen sulfide (H2S) donor GYY4137 on intracellular lipid breakdown in primary hepatic steatosis mice. Methods: In vitro model of hepatic steatosis induced by oleic acid was established. Primary hepatocytes from C57BL / 6 mice were isolated by two-step in situ perfusion and divided into four groups: the control group was cultured in normal medium for 54 h; the model group was treated with 1.2 mmol / L oleic acid (dissolved in 10% BSA) The cells were cultured for 48 h in RPMI-1640 culture medium. Cells in H2S group and DL-propargylglycine (PAG; cystathionine γ-lyase inhibitor, H2S inhibition) / L oleic acid for 48 h, then cultured in RPMI-1640 medium without phenol red (containing 1 mmol / L GYY4137 and 200 μmol / L PAG) for 6 h. Cell glycerol release and intracellular hormone-sensitive lipase (HSL) protein expression were measured. Results: Compared with the model group, the release of glycerol and the expression of phosphorylated HSL (p-HSL) in H 2 S group were significantly decreased, while the PAG group was significantly increased. CONCLUSION: GYY4137 may reduce intracellular lipidolysis in mouse primary steatosis hepatocytes by inhibiting the phosphorylation of HSL.