Effect of apolipoprotein M on the expression of sphingosine-1-phosphate receptor 1

doi:10.3877/cma.j.issn.2095-9605.2017.02.008

Abstract

Abstract:

Objective

To investigate the effect of apolipoprotein M (apoM) on the expression of sphingosine-1-phosphate receptor 1 (S1P1).

Methods

In the vitro experiment, lentiviruses with apoM gene (the experimental group) and without apoM gene (the control group) were used to infect the endothelial cells of human umbilical vein (EA.hy926 cells). Then the total mRNAs and proteins were obtained from the cell lines with stable expression, and RT-PCR was used to detect the expression level of apoM, RT-PCR and Western Blot were used to detect the expression level of S1P1 receptor. In the vivo experiment, 8~10 week-old and 25-gram-weight healthy wild-type apoM gene (apoM^+/+, the experimental group)and apoM knockout (apoM^-/-, the control group) C57BL/6 mice were selected, with 8 mice in each group. After they were sacrificed, the liver mRNAs were extracted from the aortas and reversely transcribed to cDNAs. Then RT-PCR was used to detect the expression level of apoM and S1P1 mRNA. The expressions levels of apoM mRNA, S1P1 mRNA and protein were compared between the experimental and control group in the vitro and vivo experiments.

Results

(1) The expression levels of S1P1 mRNA and protein of the EA.hy926 cells in the experimental group were significantly higher than those of the EA.hy 926 cells in the control group (P<0.05); (2) the expression levels of S1P1 mRNA and protein in the aorta of apoM^+/+ mice were significantly higher than those of the apoM^-/- mice (P<0.05).

Conclusion

ApoM can increase the expression level of S1P1 mRNA and protein in vivo and vitro.

Key words: Apolipoprotein M, Sphingosine-1-phosphate receptor 1, MRNA, Receptor expression

Min Wang, Guanghua Luo, Youpu Zhang, Bo Huang, Yang Yu, Hong Liu, Ning Xu, Xiaoying Zhang. Effect of apolipoprotein M on the expression of sphingosine-1-phosphate receptor 1[J]. Chinese Journal of Obesity and Metabolic Diseases(Electronic Edition), 2017, 03(02): 91-96.

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References 16

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基因名称	序列（5’→3’）
人ApoM	正义引物：CTGACAACTCTGGGCGTGGAT
?	反义引物：TGTCCACAGGGTCAAAAGTTGC
?	探针：FAM-AGTTCCCAGAGGTCCACTTGGGCCA-BHQ1
人S1P1	正义引物1：GGGCTCTCCGAACGCAAC
?	正义引物2：GGACCCCGACTCGAGCTG
?	反义引物：GTTCGATGAGTGATCCAGGCTT
?	探针：FAM-TCCGAGGCCCTCTCCAGCCAA-BHQ1
鼠S1P1	正义引物：CAGCTTCGTCCGGCTTGAG
?	反义引物：GTTACAGCAAAGCCAGGTCAGC
?	探针：FAM-AGGCTGCTGTTTCTCGGAGGCCTC-BHQ1
人GAPDH	正义引物：GGAAGGTGAAGGTCGGAGTC
?	反义引物：CGTTCTCAGCCTTGACGGT
?	探针：CY5-TTTGGTCGTATTGGGCGCCTG- BHQ2
鼠GAPDH	正义引物：TCTTGTGCAGTGCCAGCCT
?	反义引物：TGAGGTCAATGAAGGGGTCG
?	探针：CY5-AGGTCGGTGTGAACGGATTTGGC- BHQ2

基因名称	序列（5’→3’）
人ApoM	正义引物：CTGACAACTCTGGGCGTGGAT
?	反义引物：TGTCCACAGGGTCAAAAGTTGC
?	探针：FAM-AGTTCCCAGAGGTCCACTTGGGCCA-BHQ1
人S1P1	正义引物1：GGGCTCTCCGAACGCAAC
?	正义引物2：GGACCCCGACTCGAGCTG
?	反义引物：GTTCGATGAGTGATCCAGGCTT
?	探针：FAM-TCCGAGGCCCTCTCCAGCCAA-BHQ1
鼠S1P1	正义引物：CAGCTTCGTCCGGCTTGAG
?	反义引物：GTTACAGCAAAGCCAGGTCAGC
?	探针：FAM-AGGCTGCTGTTTCTCGGAGGCCTC-BHQ1
人GAPDH	正义引物：GGAAGGTGAAGGTCGGAGTC
?	反义引物：CGTTCTCAGCCTTGACGGT
?	探针：CY5-TTTGGTCGTATTGGGCGCCTG- BHQ2
鼠GAPDH	正义引物：TCTTGTGCAGTGCCAGCCT
?	反义引物：TGAGGTCAATGAAGGGGTCG
?	探针：CY5-AGGTCGGTGTGAACGGATTTGGC- BHQ2

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