小蘖碱对甲状腺相关眼病眼眶前脂肪细胞分化的影响

Effects of berberine on differerntiation of orbital preadipocytes of TAO patients in vitro

目的观察小蘖碱对体外培养的甲状腺相关眼病患者眼眶前脂肪细胞诱导分化的影响。方法体外培养甲状腺相关眼病患者眼眶前脂肪细胞,鉴定与传代,加入小蘖碱使其浓度分别为10 mol·L^(-1)、20 mol·L^(-1)、40 mol·L^(-1)、80 mol·L^(-1)组,另设不加小蘖碱的为对照组,进行诱导分化及油红O染色,图像分析系统计算分化后脂肪细胞内脂质/核的平均积分光密度值(integrated optical density,IOD)。结果体外培养的前脂肪细胞经免疫组织化学染色鉴定为间叶来源。不同浓度小蘖碱分化后脂肪细胞内脂质/核的平均IOD值呈浓度依赖性降低,其中对照组IOD值为1.11±0.21,10 mol·L^(-1)、20 mol·L^(-1)、40 mol·L^(-1)、80 mol·L^(-1)小蘖碱组IOD值分别为0.93±0.15、0.68±0.13、0.31±0.09、0.19±0.05,与对照组相比差异有统计学意义(F=22.53,P<0.05),组间两两比较差异均有统计学意义(均为P<0.05)。结论小蘖碱可以抑制眼眶前脂肪细胞的分化和细胞内脂质的形成,抑制程度呈浓度依赖性。

Objective To investigate the effects of berberine on the differentiation of orbital preadipocytes in the patients with thyroid associated ophthalmopathy（TAO） in vitro.Methods Orbital preadipocytes were cultured in vitro and differentiated cells were identified.The subcultured preadipocytes were incubated with different concentrations of berberine（10 mol · L^-1,20 mol · L^-1,40 mol · L^-1,80 mol · L^-1）,and with no-berberine as control.The integrated optical density（IOD） of cytoplasm/nucleolus in differentiated adipocytes was measured by oil red staining to evaluate the effects of different concentrations of berberine on lipid formation of orbital adipocytes.Results Cultured preadipocytes in vitro were identified as mesenchymal origin by immunohistochemical staining.After different concentrations of berberine differentiated fat cells,the average IOD of cytoplasm/nucleolus was decreased in a concentration dependent manner.The average optical density of cytoplasm/nucleolus in control group was 1.11±0.21,the 10 mol· L^-1,20 mol·L^-1,40 mol· L^-1,80 mol· L^-1 berberine group were respectively 0.93±0.15,0.68 ±0.13,0.31 ±0.09 and 0.19 ±0.05.Compared with the control group,the difference was statistically significant（F =22.53,P〈0.05）,there were also statistical differences compared to each other in the berberine group（all P〈0.05）.Conclusion Berberine may suppress the differentiation and lipid formation of TAO orbital preadipocytes in vitro in a concentration dependent manner.