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| Regulatory Effect of Chicken (Gallus gallus) THRSP Gene on Lipid Deposition in LMH Cells |
| LIU Shi-Hao, WEI Zhi-Heng, JIANG Xin-Cheng, YAN Yao-Hua, XU Lu, YU Jian-Feng, GU Zhi-Liang* |
| School of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, China |
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Abstract Thyroid hormone responsive spot 14 (THRSP) is a transcriptional regulator involved in the regulation of lipid metabolism in tissues. In order to understand the role of THRSP gene in lipid metabolism and its molecular mechanism, this study determined the expression of THRSP gene in 12 kinds of chicken (Gallus gallus) tissues, amplified the full-length CDS sequence of THRSP gene using the reverse transcription product of total RNA from chicken adipose tissue as a template, and constructed the eukaryotic expression vector of THRSP gene. After the THRSP gene was overexpressed and down-regulated in LMH (Leghorn male hepatoma) cells, the effect of lipid drop deposition was detected by oil red O staining, the content changes of total cholesterol (TC) and total triglyceride (TG) were determined using colorimetry method, and the expression changes of lipid metabolism-related genes were determined by qRT-PCR. The results showed that THRSP gene was highly expressed in adipose tissues and liver. Western blot analysis showed that THRSP-Myc fusion protein was successfully expressed in LMH. It was found that overexpression of THRSP gene could promote lipid drop deposition in LMH cells (P<0.001), TC (P<0.05) and TG (P<0.01) content significantly increased, the gene expression of peroxisome proliferator activated receptor γ (PPARγ)(P<0.01) and diacylglycerol acyltransferase 2 (DGAT2)(P<0.05) were up-regulated, and the gene expression of propionyl-CoA carboxylase subunit α (PCCA)(P<0.05) and phosphoenolpyruvate carboxykinase 1 (PCK1)(P<0.001) were down-regulated. When THRSP gene was knocked down in LMH cells, lipid drop deposition were inhibited (P<0.05), TC and TG contents were decreased (P=0.06), DGAT2 gene expression was down-regulated (P<0.05), while PCCA (P<0.01) and PCK1 (P<0.001) gene expression were up-regulated. The above results indicate that THRSP gene might play a role in promoting cellular lipid deposition by regulating the gene expression of PPARγ, DGAT2, PCCA, and PCK1. This study provides a new reference for further exploring the function of chicken THRSP gene.
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Received: 25 June 2023
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Corresponding Authors:
* Zhilianggu88@hotmail.com
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