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IFN -γ on pig fat cell proliferation, differentiation and lipolysis -related gene expression

Author: WangBo
Tutor: YangGongShe
School: Northwest University of Science and Technology
Course: Zoology
Keywords: IFN-γ Pig Adipocyte Proliferation Differentiation Lipolysis
CLC: Q4
Type: Master's thesis
Year: 2007
Downloads: 95
Quote: 0
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Abstract


Adipose tissue is a mammalian body energy storage place, the energy metabolism in the body play an important role, with further research, it was found that fat tissue also has the function of the endocrine and immune regulation, but the relationship between the immune system has not yet clear, therefore, this experiment 3-7 day old Landrace pigs as experimental animals to cultured preadipocytes as the research object, the immune cytokine interferon-γ (interferon-γ, IFN-γ) precursor processing fat cells by MTT colorimetric, oil red O staining extraction, semi-quantitative RT-PCR and Western blotting method, we study its pig fat precursor cell proliferation and differentiation, and the impact of lipolysis. The results obtained are as follows: 1. Detect IFN-γ on porcine preadipocytes proliferation. MTT assay was used to detect the effect of different concentrations of IFN-γ, its pig fat precursor cell proliferation effect. The results showed that, 100 IU / mL and 1000 IU / mL of IFN-γ significantly inhibited the proliferation of preadipocytes (p lt; 0.05), inhibitory effect was time-and dose-dependent; while 10 IU / mL of IFN-γ on proliferation compared with the control group in dealing with the first two days and four days the difference was not significant after 6, 8 and 10 days difference was significant (p lt; 0.05). (2) Research IFN-γ on porcine preadipocytes differentiation. By oil red O staining extraction, morphological observation, detection of IFN-γ on adipocyte triglyceride accumulation and changes in the amount of precursor fat cells fat percentage charge, charge level and morphological changes in lipid effects, the results show 10 IU / mL, 100 IU / mL and 1000 IU / mL of IFN-γ were significantly inhibited porcine preadipocytes differentiation (p lt; 0.05), the inhibition was dose-dependent; by semi-quantitative RT-PCR. IFN-γ on porcine preadipocytes differentiation marker genes LPL and PPARγ2 expression of the transcriptional level, the results show that the differentiation process can be reduced IFN-γ expression of LPL and PPARγ2 mRNA; immunoblotting (Western blot) detection of IFN- γ on PPARγ protein expression, the results show, IFN-γ can down PPARγ1 and PPARγ2 expression. 3 of IFN-γ on porcine adipocyte lipolysis related base expression. Semi-quantitative RT-PCR using technical analysis 100 IU / mL and 1000 IU / mL two concentrations of IFN-γ on adipocyte lipolysis-related genes ATGL, TGH and HSL mRNA expression; showed mature porcine adipocytes 100 IU / mL and 1000 IU / mL of IFN-γ can induce ATGL, TGH and HSL mRNA expression, 1000 IU / mL of IFN-induced significantly higher than 100 IU / mL (p lt; 0.05), at the same time, these three genes of different sensitivity to IFN-γ, wherein, HSL most sensitive, followed by TGH, ATGL least sensitive. These results show that, IFN-γ inhibited porcine adipocyte proliferation and differentiation and promote lipolysis.

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