GYY4137对小鼠原代肝细胞胞质内脂质分解的影响

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。

Effect of GYY4137 on cytosolic lipid decomposition in mouse primary hepatocytes

AIM: To evaluate the effect of GYY4137, a novel hydrogen sulfide (H₂S) donor, on cytosolic lipid decomposition in mouse primary steatosis hepatocytes. METHODS: Oleic acid (OA) was used to induce hepatic steatosis model in vitro. The C57BL/6 mouse primary hepatocytes isolated and cultured by 2-step in situ perfusion were divided into 4 groups:the cells in control group were incubated with normal medium for 54 h; the cells in model group were incubated with OA at 1.2 mmol/L for 48 h followed by serum-free phenol red-free RPMI-1640 for 6 h; the cells in H₂S group or DL-propar-gylglycine (PAG; an inhibitor of cystathione γ-lysase, inhibiting H₂S synthesis) group were incubated with OA at 1.2 mmol/L for 48 h followed by serum-free phenol red-free RPMI-1640 which contained 1 mmol/L GYY4137 or 200 μmol/L PAG for 6 h. The glycerin release and the protein expression of hormone-sensitive lipase (HSL) in the cells were mea-sured. RESULTS: Compared with model group, the glycerin release and the protein expression of phosphorylated HSL (p-HSL) in H₂S group decreased significantly, while those increased significantly in PAG group. CONCLUSION: In steatosis hepatocytes, exogenous H₂S possibly decreases cytosolic lipid decomposition by decreasing the protein level of p-HSL.