Yi Wenshu, Xu Xueliang
Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha 410008, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2010 May;35(5):511-7. doi: 10.3969/j.issn.1672-7347.2010.05.017.
To investigate the effect of mevastatin (Mev) on the expression of peroxisome-proliferator-activated receptor-gamma (PPAR-gamma) mRNA and differentiation of Thyroid-associated ophthalmopathy (TAO) derived orbital preadipocytes in vitro.
Orbital 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 micromol/L (B1), 10 micromol/L(B2),20 micromol/L (B3) mevastatin respectively during the whole process of differentiation. The differentiation of orbital preadipocytes in Group B4 and B5 was interfered with 10 micromol/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-gamma mRNA was detected by reverse transcription polymerase chain reaction.
The light absorption value (A) and PPAR-gamma 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-gamma 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.
Mevastatin inhibits the differentiation of TAO derived orbital preadipocytes by blocking PPAR-gamma 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.
探讨美伐他汀(Mev)对体外培养的甲状腺相关性眼病(TAO)来源的眼眶前脂肪细胞过氧化物酶体增殖物激活受体γ(PPAR-γ)mRNA表达及分化的影响。
取行眼眶减压手术的TAO患者眼眶脂肪组织。将从眼眶脂肪组织培养的眼眶前脂肪细胞分为A组(对照组)和B组(干预组)。B组再分为B1-B5组,所有组均用混合分化培养基诱导分化为成熟脂肪细胞。分化全过程为10天。A组眼眶前脂肪细胞用常规诱导剂诱导分化,而B1、B2和B3组在分化全过程分别用5 μmol/L(B1)、10 μmol/L(B2)、20 μmol/L(B3)美伐他汀干预。B4和B5组眼眶前脂肪细胞在分化过程的第4天(B4)或第8天(B5)用10 μmol/L美伐他汀干预直至分化结束。用油红O染色法测定分化后脂肪细胞内脂肪蓄积情况。用酶联免疫吸附测定法在492 nm处测定光吸收值。用逆转录聚合酶链反应检测PPAR-γ mRNA的表达。
A组、B1组、B2组和B3组分化细胞的光吸收值(A)及PPAR-γ mRNA表达依次降低,A组与B1组、B2组、B3组中任意两组间差异均有统计学意义(P<0.05)。A组、B4组和B2组分化细胞的A值及PPAR-γ mRNA表达依次降低,这3组中任意两组间差异均有统计学意义。然而,A组与B5组间差异无统计学意义。
美伐他汀通过阻断PPAR-γ mRNA表达抑制TAO来源的眼眶前脂肪细胞分化。抑制程度不仅呈浓度依赖性,还与分化阶段有关。分化越早,抑制越强。