【作者】 张毅； 刘精东； 林安华；

【Author】 Zhang Yi~1,Liu jing-dong~2,Lin An-hua~2 (1.2007 Grade of Medical Department of Grade School of Nanchang University; 2.Department of Endorcrine Disease,Jiangxi Province People’s Hospital)

【机构】 南昌大学研究生院医学部； 江西省人民医院内分泌科；

【摘要】 目的探讨噻唑烷二酮类药物对脂肪细胞增殖、分化及内分泌功能的影响。方法不同浓度的吡咯列酮的基础培养液体外培养前脂肪细胞,48小时后用ELISA法检测细胞上清液apelin浓度,并确定有效的吡格列酮干预浓度。设立空白对照组A、分化对照组B、实验组C三组,在干预后的第24、48、72小时分别检测各组apelin浓度,并观察细胞增殖、聚脂分化指标。后续于分化的第5、10天,测定三组细胞油红O染色吸光度值。结果 A、B及C组上清液中可检测到apelin,其结果随培养时间而变化,B组、C组培养72小时后apelin浓度较A组分别增加了33.7%、32.5%（P<0.05）。2、对细胞分化的影响:油红O染色提取法测定结果表明,当吡格列酮浓度分别为10μmol·L^-1,100μmol·L^-1时其吸光度值大于对照组。在第24、48小时,C组油红O染色吸光度值与B组比较无明显差异;第72小时,C组吸光度值高于A组,差异有统计学意义（P<0.05）,但低于B组;而在第10天B、C两组差异不明显（P≥0.05）,但与A组比较,其差异仍较明显（P<0.05）。3、对细胞增殖的影响:MTT法检测结果显示,随吡格列酮浓度增加,前脂肪细胞数有增加趋势,100μmol·L^-1吡格列酮明显促进前脂肪细胞的增殖,在24、48、72小时细胞增殖较对照组分别增加了8.1%、20.2%和15.2%（P<0.05）。结论前脂肪细胞可分泌脂肪因子apelin,其表达水平与细胞生长状态有关。噻唑烷二酮可促进前脂肪细胞增殖和分化,促增殖作用呈时间和剂量依赖性,其促分化作用具有剂量依赖趋势。噻唑烷二酮可通过调节前脂肪细胞的生长而影响其分泌功能。更多还原

【Abstract】 Objective To observe the process of preadipocyte proliferation and differentiation,investigate the mechanisms of thiazolidinedione affect endocrine function of adipocyte.Method:1.Preadipocytes were serperately cultured by the basic medium with four different consistences of pioglitazone in vitro,then detected apelin concentration of the cell supernatant by ELISA after 48 hours,and evaluated cell proliferation by MTT test,to find an effective concentration of pioglitazone.According the different medium,the preadipocyte devided into three groups including control group A,differentiation group B,experimental group C.At the varied experimental phases of 24,48,72 hours,the apelin concentration of supernatant was detected,the cell parameters of proliferation and differentiation were observed.In 5 and 10 days,the absorbance of cells stained with oil red O was detected.The apelin concentrations,and parameters of preadipocyte proliferation and differentiation of the three groups were evaluation.Results 1.Apelin could be detected in the supernatant of A,B and C group, and its concentration was changed with the culture time,at 72 hours the apelin levels in B,C group remarkly increased by 33.7%,32.5%than A group（P<0.05）.2.The effect on cell differentiation:oil red O staining test showed that the absorbance values of the 10μmol ?L^-1,l00μmol·L^-1 pioglitazone groups were greater than the control group.At 24,48 hours,the absorbance of C group staining with oil red O had no significant difference compared with B group;At 72 hours,the absorbance value of C group was higher than the value of A group,the difference was statistically significant（P<0.05）, but lower than B group;in the first 10 days,the difference between B and C group was not significant（P≥0.05）,but the difference was still significant between A and C group（P<0.05）.3.The effect on cell proliferation:MTT test assay showed that with pioglitazone concentration,pre-fat cells to an increasing trend,l00μmol ? L^-1 pioglitazone significantly promote preadipocyte proliferation.In 24,48,72 hours,the result of cell proliferation increased by 8.1%,20.2%and 15.2%compared with control group（P<0.05）.Conclusion Adipocytokine as apelin can be secreted in preadipocyte, and its secretion is related with cell growth state.Thiazolidinedione can promote preadipocyte proliferation and differentiation,promote cell proliferation in time and dose dependent manner,its role in promoting differentiation in a dose dependent manner.Thiazolidinedione can affect the secretion function of preadipocyte by regulating the state of growth.更多还原