Regulation of the expression of porcine angiotensin converting enzyme 2 by polyunsaturated fatty acids
Date Issued
2009
Date
2009
Author(s)
Tseng, Yu-Wen
Abstract
Using suppression subtractive hybridization technique, we found that two unkown genes (AEUG1 and AEUG3) were highly expressed in the adipocytes compared with preadipocytes. We then identified that these two genes were indicated to be angiotensin converting enzyme 2 (ACE2). We applied 3’RACE (rapid amplified of cDNA end), 5’RACE, and PCR based on primers from full length cDNA sequences of other species published on NCBI, to obtain the full length porcine ACE2 cDNA sequence.olyunsaturated fatty acids (PUFA) are precursors of 3 series of eicosanoids which are antiaggregators and vasoconstrictors. Because the functions of eicosanoids link PUFA to regulating blood pressure, we hypothesize that PUFA can regulate the expression of the vasodilator, ACE2. Preadipocytes from Landrace pigs were induced to differentiate for 4 days, then treated with 50 μM of different PUFA, conjugted linoleic acid (CLA), arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), or stearic acid (18:0). Addition of either EPA or 18:0 for 48 h did not change the ACE2 mRNA level, whereas the treaments of AA、CLA and DHA significantly decreased ACE2 mRNA level after 48 h. To further understand how PUFA metabolites affect ACE2 mRNA expression, we inhibited individual enzymes that involved in the eicosanoid production. The inhibitors for monooxygenase, lipooxygenase, and cyclooxygenase are clotrimazole, ETYA, and indomethacin, respectively. We found all three inhibitors could recover ACE2 expression inhibition by AA treatment, suggesting that three eicosanoids are involved in regulating the expression of ACE2. To further investigate the function of ACE2, we enhanced ACE2 expression in porcine adipocyte by using lentivirus as a vector. After overexpression of ACE2 in preadipocytes, we induced preadipocytes to differentiate into adipocytes for 3 days, then analysed vifferentiation marker genes (CCATT/enhancer binding protein β、sterol regulatory element binding protein-1C and peroxisome proliferator-activated receptor γ) and lipolytic genes (adipose triglyceride lipase and hormone sensitive lipase). The results showed the expression of both differentiation marker genes and lipolytic genes were not modified by overxpression ACE2. Therefore ACE2 may not have a role in regulating early stage adipocyte differentiation and lipolysis.n conclusion, we found PUFA could down-regulate the expression of ACE2 through its metabolites in porcine adipocytes. ACE2 may not have a function on modifying the expression of early stage adipocyte differentiation gene and lipolytic genes. Taken together, PUFA can down-regulate ACE2 gene expression but the physiological modification by which ACE2 mediating requires further demonstration.
Subjects
Angiotensin converting enzyme 2
polyunsaturated fatty acids
porcine adipocyte
Type
thesis
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