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| Effect of Down-regulated Perilipin 1 Gene Expression on Lipolysis of 3T3-L1 Adipocytes |
| ZHAO Zhi-wu, WANG Jun-shi, MA Min, ZHANG Shao-hua, YAN Jiong |
| School of Public Health, Shanxi Medical University, Taiyuan 030001, China |
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Abstract Objective:To investigate the effect on lipolysis of 3T3-L1 adipocytes by down-regulating Perilipin 1 gene expression. Methods: Three shRNA recombinant vectors targeting mouse PLIN1 gene and one negative control were designed, synthesized and detected by bacterium liquid PCR reaction and DNA sequencing. Western blot analysis was performed to verify the transfection effects.Bodipy 493/503 staining was used to observe the lipid droplets size, and the content of cellular triglyceride and glycerol were detected after transfection of adipocytes. The adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL) and phosphorylase HSL (p-HSL) were detected by Western blot, and the cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) were measured by enzyme linked immunosorbent assay (ELISA). Results: Every recombinant vector was constructed successfully, and the three positive vectors could significantly inhibit the protein expression of PLIN1A (P<0.05). Knockdown of PLIN1 gene expression decreased lipiddroplets size and cellular triglyceride level and increased glycerol level. In addition, the level of ATGL and HSL was increased significantly (P<0.05), and the level of cellular cAMP and PKA had no obvious change (P>0.05). Conclusion: Down-regulating PLIN1 gene could promote lipolysis in 3T3-L1 adipocytes by up-regulating the level of ATGL and HSL, and cAMP-PKA signaling pathway had no significant effect on it.
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Received: 17 September 2015
Published: 09 December 2015
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