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用噻唑烷二酮使猪卫星细胞转分化为脂肪母细胞。

Transdifferentiation of porcine satellite cells to adipoblasts with ciglitizone.

作者信息

Singh N K, Chae H S, Hwang I H, Yoo Y M, Ahn C N, Lee S H, Lee H J, Park H J, Chung H Y

机构信息

Product and Utility Division, National Livestock Research Institute, Suwon 441-350, Republic of Korea.

出版信息

J Anim Sci. 2007 May;85(5):1126-35. doi: 10.2527/jas.2006-524. Epub 2006 Dec 18.

Abstract

Ciglitizone, a class of thiazolidinediones, acts as a potent activator of the adipose differentiation program in established preadipose cell lines. Thiazolidinediones have also been investigated in diabetic patients and have been reported to act as peroxisome proliferator-activated receptor-gamma ligands. Intramuscular adipogenesis or marbling through transdifferentiation of satellite cells in cattle was successfully conducted earlier. In this report, the effects of ciglitizone on the differentiation pathway of porcine myogenic satellite cells was investigated. Semitendinosus muscle was aseptically taken from 10-d-old piglets under general anesthesia, and porcine satellite cells were obtained and grown to near confluence. Postconfluent cells (d 0) were further cultured in differentiation medium containing an adipogenic mixture plus ciglitizone (10 microM) for 48 h. From d 2 onward, the cells were cultured only in the presence of ciglitizone until d 10. Controls were cultured in differentiation medium only. Exposure of porcine satellite cells to the adipogenic mixture plus ciglitizone generated lipid droplets on d 2, and subsequently, exposure of cells to ciglitizone alone helped in cytoplasmic lipid filling, providing them with the acquisition of adipocyte morphology. An increase (P < 0.05) in the fusion (structures containing 2 to 3 nuclei) of satellite cells was observed, and myosin heavy chain appeared with greater intensity (immunohistochemistry) in the control group from d 2 onward. Adipocyte-specific transcriptional factors (i.e., CCAAT/enhancer binding protein-alpha and peroxisome proliferator-activated receptor-gamma) were predominant during transdifferentiation and were observed with immunohistochemistry, Western blot (approximately 47.2 and approximately 60.4 kDa, respectively), and real-time PCR. Ciglitizone appeared to convert the differentiation pathway of satellite cells into that of adipoblasts.

摘要

噻格列酮是噻唑烷二酮类药物的一种,在已建立的前脂肪细胞系中,它是脂肪分化程序的强效激活剂。噻唑烷二酮类药物也已在糖尿病患者中进行了研究,据报道其作为过氧化物酶体增殖物激活受体γ配体发挥作用。早期已成功实现牛卫星细胞通过转分化进行肌肉内脂肪生成或大理石花纹状脂肪沉积。在本报告中,研究了噻格列酮对猪成肌卫星细胞分化途径的影响。在全身麻醉下,从10日龄仔猪无菌采集半腱肌,获得猪卫星细胞并培养至接近汇合状态。汇合后的细胞(第0天)在含有脂肪生成混合物加噻格列酮(10微摩尔)的分化培养基中进一步培养48小时。从第2天起,细胞仅在噻格列酮存在的情况下培养至第10天。对照组仅在分化培养基中培养。猪卫星细胞暴露于脂肪生成混合物加噻格列酮后,在第2天产生脂滴,随后,细胞单独暴露于噻格列酮有助于细胞质脂质填充,使其获得脂肪细胞形态。观察到卫星细胞融合(含有2至3个核的结构)增加(P<0.05),并且从第2天起,对照组中肌球蛋白重链出现的强度更高(免疫组织化学)。脂肪细胞特异性转录因子(即CCAAT/增强子结合蛋白α和过氧化物酶体增殖物激活受体γ)在转分化过程中占主导地位,并通过免疫组织化学、蛋白质印迹法(分别约为47.2 kDa和约60.4 kDa)和实时PCR观察到。噻格列酮似乎将卫星细胞的分化途径转变为脂肪母细胞的分化途径。

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