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| Influence of Interfering CAT Gene by siRNA on Differentiation of Porcine (Sus scrofa) Subcutaneous Preadipocytes |
| SHAN Bao-Sen1, 2, LIU Xin1, LUO Wu1, SHAO Yong-Gang3, WEI Wei1, CHEN Jie1, ZHANG Li-Fan1, 2, * |
1 College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China;
2 National Experimental Teaching Demonstration Center of Animal Science, Nanjing Agricultural University, Nanjing 210095, China;
3 College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China |
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Abstract As one of the major factors for affecting the growth and meat quality of pigs (Sus scrofa), subcutaneous fat deposition has always been a hot issue in pig production. Catalase (CAT) is a key antioxidant enzyme in the oxidative metabolism process and plays vital roles in the fat deposition of animals. To investigate the role of CAT gene in the porcine subcutaneous preadipocytes differentiation, the expression patterns of CAT during the preadipocyte differentiation process and CAT mRNA levels in various tissues of Erhualian pigs were detected using qRT-PCR. Moreover, CAT gene was knockdowned by small interfering RNA (siRNA) fragments, and then, the ability of subcutaneous preadipocytes differentiation was assessed via oil red O staining. The expression levels of important candidates in siCAT treated adipocytes, including the key adipocyte differentiation-related genes such as peroxisome proliferator activated receptor γ gene (PPARγ), CCAAT enhancer binding protein α gene (C/EBPα), fatty acid binding protein 4 gene (FABP4), apolipoprotein E gene (ApoE), adiponectin, C1Q and collagen domain containing gene (ADIPOQ), perilipin 1 gene (PLIN1), and hormone-sensitive lipase gene (HSL), and the antioxidant-related genes such as superoxide dismutase 2 gene (SOD2), glutathione synthetase gene (GSS), and peroxidase 3 gene (PRDX3) were detected by qRT-PCR. The results indicated that CAT gene was mainly expressed in subcutaneous adipose, kidney, and liver tissues of pigs and its expression levels in subcutaneous adipose tissues were significantly higher than that in intramuscular adipose tissues (P<0.01). During the differentiation process, the expression levels of CAT gene increased significantly at different differentiation stages, with the highest expression levels on the 4th day of the preadipocytes differentiation. Silencing of CAT increased the contents of hydrogen peroxide (H2O2) (P<0.01) and also decreased the triglyceride levels of subcutaneous preadipocytes (P<0.05). Furthermore, interfering CAT expression reduced the expression levels of ADIPOQ (P<0.05) and PLIN1 (P<0.01) and simultaneously increased the levels of HSL gene (P<0.05), with non-significant effect on the expression levels of PPARγ, C/EBPα, FABP4, ApoE, SOD2, GSS, and PRDX3 genes (P>0.05). Taken together, this study demonstrated CAT gene knockdown could inhibit the differentiation and lipid deposition of subcutaneous preadipocytes of Erhualian pigs, which might result from downregulating lipogenesis-related genes of ADIPOQ and PLIN1 and simultaneously upregulating the expressions of lipolysis-related HSL gene. These results could provide new evidence and ideas for further revealing the mechanism of CAT gene on adipogenesis and reducing the subcutaneous fat deposition of pigs.
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Received: 30 October 2018
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Corresponding Authors:
lifanzhang@njau.edu.cn
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