摘要
目的研究平目汤对Graves眼病眼眶脂肪细胞FAS、FAS L、CHOP、CASPASE-4蛋白表达的影响,探讨眼眶成熟脂肪细胞凋亡的机制。方法体外培养Graves眼病患者眼眶前脂肪细胞,传代并诱导其分化为成熟脂肪细胞。平目汤含药血清,配成终浓度分别为5%、10%、20%的平目汤低、中、高剂量组。采用MTT法检测前脂肪细胞增殖情况。Western blotting法检测平目汤对眼眶成熟脂肪细胞FAS、FAS L、CHOP及CASPASE-4蛋白表达的影响。结果 (1)MTT法检测结果显示,与空白对照组比较,平目汤低、中、高剂量组的生长曲线显著下移,其中平目汤中剂量组活性下降趋势最明显。地塞米松组与空白对照组差异无统计学意义(P>0.05)。相同的时间点与空白对照组比较,平目汤各组OD值均减小,其中以平目汤中剂量组降低最为明显,差异有统计学意义(P<0.05)。(2)Western blotting结果显示:与空白对照组比较,除地塞米松组外,各组FAS、FAS L蛋白表达均上调,差异有统计学意义(P<0.05),其中FAS蛋白表达以平目汤中剂量组最高,其次为低剂量组,FAS L蛋白表达以平目汤高剂量组最高,其次为中剂量组,但二者比较差异无统计学意义(P<0.05)。与空白对照组比较,除地塞米松组外,平目汤各组CHOP、CASPASE-4蛋白表达均上调,差异有统计学意义(P<0.05),其中平目汤各组组间比较CHOP蛋白表达以高剂量组最高,其次为中剂量组;CASPASE-4蛋白表达以平目汤中剂量组最高,其次为低剂量组。结论平目汤通过死亡受体途径和内质网途径诱导FAS、FAS L、CHOP、CASPASE-4活化,促进成熟脂肪细胞凋亡,进而减少眼眶脂肪细胞积聚,发挥治疗效果。
Objective To study the effects of medicated serum of Pingmu Decoction on protein expressions of FAS,FAS L,CHOP and CASPASE-4 in orbital adipocytes in Graves' ophthalmopathy,and discuss the mechanism of apoptosis in orbital mature adipocytes. Methods The orbital preadipocytes from patients with Graves' ophthalmopathy were cultured in vitro,and differentiated into the mature adipocytes after cell passage and induction. The medicated serum of Pingmu Decoction were prepared,which divided into the low-,medium-and high-dose group with the final concentrations as 5%,10% and 20% respectively. The proliferation of preadipocytes was detected by MTT assay. The effects of medicated serum of Pingmu Decoction on protein expressions of FAS,FAS L,CHOP and CASPASE-4 in orbital mature adipocytes were detected by Western blotting. Results(1)The MTT assay showed that,compared with the blank control group,the growth curve was significantly downward in the Pingmu Decoction groups with low-,medium-and high-dose,and the decrease trend of activities in the medium-dose group was the most significant. There was no statistically significant difference between the dexamethasone group and blank control group( P > 0. 05). Compared with the blank control group at the same timepoint,the OD value was decreased in the Pingmu Decoction groups with low-,medium-and high-dose,and the decrease in the medium-dose group was the most significant with statistical difference( P < 0. 05).(2)The results of Western blotting showed that,except the dexamethasone group,compared with the blank control group,the protein expressions of FAS and FAS L were up-regulated in the Pingmu Decoction groups with different doses,with statistical differences( P < 0. 05). The protein expression of FAS was highest in the medium-dose group,followed by low-dose group; the protein expression of FAS L was highest in the high-dose group,followed by medium-dose group,but there was no statistically significant difference between these two groups. Except the dexamethasone group,compared
出处
《上海中医药杂志》
2017年第9期85-89,共5页
Shanghai Journal of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(81373617)
上海市科委资助项目(15401930400)
上海市浦东新区名中医工作室建设资助项目(PDZYXK-3-2014005)
