类维甲酸X受体α (RXRα)对原代培养猪前脂肪细胞凋亡的影响。

Lin Ya-qiu, Zhuang He-Lin, Yang Gongshe
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引用次数: 0

摘要

采用半定量逆转录聚合酶链式反应(半qrt - pcr)、细胞转染、Hoechst 33342、AO和流式细胞术等方法,分别研究了9-顺式维甲酸(9-cis- RA)类视黄醇X受体α(RXRα)配体转染增效绿色荧光蛋白C2-RXRα (pEGFPC2-RXRα)和RXRα-小干扰RNA (RXRα- sirna)后猪前脂肪细胞凋亡的变化。观察了猪前脂肪细胞凋亡的形态学变化。细胞先是收缩,染色质收缩,细胞核缩缩至致密体,然后分裂,核膜完整,不断出芽和脱落,最后细胞变成几个大小不等的膜包裹的凋亡小体。与对照组相比,10 μmol/L 9-cisRA处理组细胞凋亡率显著降低,10 μmol/L 9-cisRA处理组细胞凋亡率显著升高(P < 0.05)。转染pEGFPC2-RXRα和RXRα- sirna后,RXRα mRNA的表达分别显著上调和下调(P < 0.05), pEGFPC2-RXRα组细胞凋亡率显著降低,RXRα- sir - NA组细胞凋亡率显著高于对照组(P < 0.05)。提示RXRα对猪前脂肪细胞凋亡有抑制作用。
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Effect of retinoid X receptor α (RXRα) on apoptosis in primary cultured porcine pre-adipocytes.
By means of semi-quantitative reverse transcription-polymerase chain reaction (semi-qRT-PCR), cell transfection, Hoechst 33342 , AO and flow cytometry methods, the apoptotic change of porcine preadipocyte treated with 9-cis retinoic acid(9-cis- RA) of retinoid X receptor α(RXRα) ligand, transfected with pEnhanced green fluorescent proteins C2-RXRα (pEGFPC2-RXRα) and RXRα-small interfering RNA (RXRα-siRNA), was investigated respectively. The morphologic change of apoptosis in treated porcine preadipocyte was observed. It appeared the cell constriction, cellular shrinkage and condensation of chromatin, pyknosis of nucleus in- to compact body firstly, and then fragmentation with completely nucleus membrane, budding and shedding continuously, at last cell turns to several apoptotic bodies enwraped with membrane in different size. Moreover, compared with control group, apoptotic rate was significantly decreased in 10 nmol/L 9-cisRA treated group, and higher in the 10 μmol/L 9-cisRA treated group (P 0.05). The expression of RXRα mRNA was significantly upregulated and downregulated when transfected with pEGFPC2-RXRα and RXRα-siRNA(P 0.05), respectively, and the apoptotic rate was significantly decreased in pEGFPC2-RXRα group while RXRα-siR- NA group significantly higher compared with control group (P 0.05). It implies that RXRα inhibits the apoptosis of porcine preadipocyte.
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