硫化氢对脂肪肝脂细胞脂联素分泌的调节

Regulatory effect of hydrogen sulfide on adiponectin secretion in fatty liver adipocytes

目的 观察硫化氢是否调节脂肪肝脂细胞脂联素的分泌。 方法 对32只雄性SD大鼠采用完全随机化法分成正常喂养组 （n=8）、 高脂饮食组 （n=8）、高脂＋半胱氨酸组 （n=8）、高脂＋炔丙基甘氨酸（胱硫醚-γ-裂解酶抑制剂）组（n=8）。利用高脂饮食复制脂肪肝动物模型，加用1%L-半胱氨酸和炔丙基甘氨酸（PPG）喂养改变硫化氢的产生，观察8周后动物肝脏脂肪变性和血清内脂联素的含量。取动物大网膜脂肪组织，利用胶原酶Ⅰ消化法分离脂肪细胞，加用NaHS和PPG，体外刺激7 d后，检测细胞上清脂联素的含量变化。结果高脂饮食8周后脂肪肝形成，应用L-半胱氨酸后脂肪变性明显减轻；而应用PPG肝脏脂肪变性加重。高脂饮食组动物血清中硫化氢和脂联素含量分别为（21.13±7.06）mmol/L和（3.16±1.15）mg/L，较正常喂养组（29.13±13.06）mmol/L和 （8.98±2.84）mg/L明显减少（P=0.0229，P=0.0062）。与高脂饮食组比较，高脂＋半胱氨酸组两者明显增加为（35.47±9.04）mmol/L和（6.54±1.38）mg/L（P=0.0032,P=0.0131）；高脂＋PPG组两者明显减少为（16.65±8.79） mmol/L 和（2.50±0.91） mg/L（P=0.0191，P=0.0021）。正常喂养组与高脂饮食组分离脂肪细胞培养液中的硫化氢含量差异无统计学意义［（26.77±12.65）比（28.76±9.09）mmol/L，P=0.0927］。与高脂饮食组比较，高脂＋硫氢化钠和高脂＋PPG组的细胞上清液中硫化氢增高和减少明显（P=0.0000，P=0.0000）。高脂饮食组与正常喂养组分离脂肪细胞培养液中的脂联素含量差异无统计学意义［（4.21±1.61）比（4.49±1.09）mg/L，P=0.1076］，加NaHS和PPG细胞上清液中脂联素较高脂饮食组上清液中明显增高和减少［（5.77±0.86） 比 （4.21±1.61）mg/L, P=0.0094;（3.01±1.26） 比 （4.21±1.61）mg/L，P=0.0108）］。 结论硫化氢能促进脂肪肝脂细胞脂联素的分泌。

Objective o observe whether hydrogen sulfide modulates adipocytes to secrete adiponectin. Methods Thirty-two male SD rats were randomly divided into normal diet group （n=8）, high-fat diet group （n=8）, high-fat ＋ cysteine group （n=8）, and high-fat ＋ propargylglycine （cystathionine-γ-lyase inhibitor） group （n=8）. The fatty liver animal model was established using the high fat diet; meanwhile, L-cysteine was applied to the animals to produce endogenous hydrogen sulfide and propargylglycine to inhibit the production of endogenous hydrogen sulfide. Fatty degeneration of the liver of animals and plasma within the adiponectin concentration was observed after 8 weeks. The omental adipose tissues were excluded and digested using collagenase I to isolate fat cells. Sodium hydrosulfide and propargylglycine was applied to stimulate the cells in vitro for 7 days before the detection of the cell supernatant adiponectin content. Results Liver steatosis was established after high-fat diet for eight weeks. Steatosis was significantly extenuated after the application of L-cysteine and aggravated by propargylglycine. Plasma hydrogen sulfide and adiponectin levels in fat diet group animals were （21.13±7.06）mmol/L and （3.16±1.15）mg/L respectively,which significantly decreased when compared with the normal diet group with the levels of hydrogen sulfide （29.13±13.06）mmol/L and adiponectin （8.98±2.84）mg/L（P=0.0229,P=0.0062）. Hydrogen sulfide [（35.47±9.04）mmol/L] and adiponectin [（6.54±1.38）mg/L] levels in the high fat ＋ cysteine group significantly increased（ P=0.0032, P=0.0131）. However, hydrogen sulfide [（16.65±8.79）mmol/L] and adiponectin [（2.50±0.91）mg/L] in high fat ＋ propargylglycine group were significantly decreased （P=0.0191, P=0.0021）. Hydrogen sulfide was not significantly different in the supernatant of adipocyte isolated from normal diet group [（26.77±12.65）mmol/L] and from high-fat diet group [（28.76±9.09）mmol/L] （P=0.0927）, but that in the sodium hydrosulfide and propargylglycine cells supernatants significantly either increased or decreased （ P=0.0000,P=0.0000）. Adiponectin in the supernatants of high-fat diet group and normal diet group adipocyte showed no significant difference [（4.21±1.61）mg/L vs （4.49±1.09）mg/L, P=0.1076）]. The levels of adiponectin in sodium hydrosulfide and propargylglycine cell supernatants increased and decreased respectively than that in higher fat diet group cells [（5.77±0.86）mg/L vs （4.21±1.61）mg/L, P=0.0094; （3.01±1.26）mg/L vs （4.21±1.61）mg/L, P=0.0108]. Conclusion Hydrogen sulfide promotes adipocytes to secrete adiponectin.