美伐他汀抑制甲状腺相关眼病眼眶前脂肪细胞的分化

目的：观察美伐他汀对体外培养的甲状腺相关眼病（thyroid associated ophthalmopathy,TAO）来源眼眶前脂肪细胞分化及过氧化物酶体增殖物激活受体 γ（peroxisome proliferator activated receptor γ,PPAR γ）mRNA表达的影响。方法：眼眶组织取自TAO行开眶减压术的患者。将TAO病眼眶前脂肪细胞分为A组（对照组）与B组（干预组）,其中B组又分为B1～B5组,均采用鸡尾酒法诱导10 d,使其分化为脂肪细胞。A组采用常规诱导剂,B1～B3组在分化全程中分别加入5 μmol/L( B1组)、10 μmol/L(B2组)、20 μmol/L(B3组)的美伐他汀,B4～B5组在分化第4天(B4组)与第8天( B5组)分别加入10 μmol/L的美伐他汀直至分化结束。分化后的细胞以油红O染色后,于酶联免疫检测仪492 nm波长处测吸光度值,以测定分化后脂肪细胞相对含量。反转录聚合酶链反应法（reverse transcription polymerase chain reaction,RT PCR）检测各组在分化结束时PPAR γ mRNA的表达水平。结果：分化后的A,B1,B2,B3组细胞吸光度值及PPAR γ mRNA表达均依次下降,组间两两比较差异均具有统计学意义（P＜0.05）;对A,B2,B4,B5组分化后细胞的吸光度值、PPAR γ mRNA表达分别进行两两比较,除A与B5组间的吸光度值、PPAR γ mRNA表达差异无统计学意义（P＞0.05）外,A,B4,B2组的吸光度值及PPAR γ mRNA表达均依次下降,组间两两比较差异均有统计学意义（P＜0.05）。结论：美伐他汀抑制体外培养的TAO眼眶前脂肪细胞的分化与PPAR γ mRNA的表达,抑制程度呈浓度依赖性,并与前脂肪细胞分化的阶段有关,越在分化的早期,抑制作用越强。

Mevastatin inhibits the differentiation of thyroid-associated ophthalmopathy derived orbital preadipocytes

ObjectiveTo investigate the effect of mevastatin（Mev）on the expression of peroxisome proliferator activated receptor γ (PPAR γ) mRNA and differentiation of Thyroid associated ophthalmopathy (TAO) derived orbital preadipocytes in vitro.MethodsOrbital adipose tissues were obtained from TAO patients undergoing orbital decompression surgery.The orbital preadipocytes cultured from the orbital adipose tissues were divided into Group A （a control group）and Group B (an intervention group ). Group B was subdivided into Group B1-B5, all groups were stimulated to differentiate into mature adipocytes with cocktail differentiation medium.The entire course of differentiation was 10 d. The differentiation of orbital preadipocytes in Group A was induced with routine inducer,while at in Group B1,B2,and B3 was interfered with 5 μmol/L( B1),10 μmol/L(B2),20 μmol/L(B3) mevastatin respectively during the whole process of differentiation. The differentiation of orbital preadipocytes in Group B4 and B5 was interfered with 10 μmol/L mevastatin day 4 (B4) or day 8 (B5) of the differentiation process until the entire course was over. Intracellular fat accumulation in differentiated adipocytes was determined by oil red O staining. The value of optical absorption was measured at 492 nm with enzyme linked immunosorbent assay. The expression of PPAR γ mRNA was detected by reverse transcription polymerase chain reaction. ResultsThe light absorption value (A) and PPAR γ mRNA expression of differentiated cells in Group A,B1,B2,and B3 decreased successively,and there was significant difference in any of the 2 groups among Group A,B1 and B2,and B3 (P＜0.05). The value A and PPAR γ mRNA expression of differentiated cells in Group A, B4, and B2 decreased successively,and the difference in any of the 2 groups among these 3 groups was significant. However, there were no significant difference between Group A and B5. ConclusionMevastatin inhibits the differentiation of TAO derived orbital preadipocytes by blocking PPAR γ mRNA expression. The degree of inhibition is not only concentration dependent but also associated with the stage of differentiation. The earlier the differentiation, the stronger the inhibition.