目的:观察obestatin对3T3-L1前脂肪细胞增殖分化的影响。方法:应用不同浓度的obestatin(10^-8、10^-9、10^-10、10^-11、10^-12mmol/L)和10^-10mmol/L ghrelin干预体外培养的3T3-L1前脂肪细胞,MTT法观察其对细胞增殖的影响,并与空白对照组比较;分别应用10^-10mmol/L obestatin和ghrelin全程干预3T3-L1前脂肪细胞分化成熟过程(分化第1-10日),采用油红O染色法鉴定脂肪细胞分化并测定分化成熟脂肪细胞的脂肪含量,RT-PCR法检测分化过程中及成熟脂肪细胞过氧化物酶体增殖物激活受体γ2(PPARγ2)mRNA的表达水平,并与对照组(加常规诱导剂)比较。结果:与空白对照组相比,不同浓度obestatin组细胞数量均明显降低(P〈0.05);10^-10mmol/L obestatin连续作用10 d的3T3-L1前脂肪细胞与对照组相比,脂肪产量明显减少(P〈0.05);脂肪细胞分化过程中PPARγ2基因表达逐渐增多;在成熟脂肪细胞中,与对照组相比,obestatin组PPARγ2基因表达显著降低(P〈0.05)。Ghrelin的作用与之完全相反。结论:Obestatin抑制3T3-L1前脂肪细胞的增殖和分化。
Objective: To investigate the inhibitory effects of obestatin on proliferation and differentiation of 3T3-L1 preadipocytes. Methods: Obestatin (10^-8 mmol/L, 10^-9 mmol/ L, 10^-10 mmol/ L, 10^-11 mmol/ L,and 10^-12 mmol/L) and ghrelin (10^-10 mmol/L) were used to treat 3T3-L1 preadipocytes. Cell proliferation was assessed by MTT assay and the results were compared with that of blank control group. The differentiation of 3T3-L1 preadipocytes (from day 1 to day 101 was interfered with obestatin or ghrelin (both at 10^-10 mmol/L). Intracellular fat accumulation in differentiated adipocytes was determined by oil red O staining and the expression of PPARγ2 mRNA was detected by RT-PCR; the results were compared with that of control group (induced with routine inducer). Results: Compared with the blank control group, obestatin-treated groups (various concentrations of obestatin) had significantly less cells(P〈0. 05). Oil red O staining revealed that, compared with control group, the formation of lipid droplets was significantly decreased after 10 days' of treatment with 10^-10 mmol/L obestatin (P〈0. 05). The expression of PPARγ2 gene increased with the progress of 3T3-L1 preadipocytes differentiation. PPARγ2 mRNA level in mature adipocytes of obestatin group was significantly lower than that in the cells of control group. The effect of ghrelin was contrary to that of obestatin. Conclusion: Obestatin can inhibit the proliferation and differentiation of 3T3-L1 preadipocyes.