Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Inhibitory Effect of Phorbol 12-Myristate 13-Acetate on NO Production Induced by Interleukin-1 beta in Aortic Vascular Smooth Muscle Cells of Rats

혈관평활근세포에서 Phorbol 12-Myristate 13-Acetate의 전처리가 Interleukin-1β에 의한 Nitrite생성에 미치는 영향

  • 윤병헌 (경북대학교 의과대학 약리학교실) ;
  • 김인겸 (경북대학교 의과대학 약리학교실) ;
  • 박태규 (경북대학교 의과대학 약리학교실) ;
  • 김중영 (경북대학교 의과대학 약리학교실)
  • Published : 2003.08.01
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Abstract

To examine the role of protein kinase C (PKC) in regulation of interleukin-1 beta (IL-1$\beta$)-induced iNOS expression, IL-1$\beta$-induced nitrite production was observed in cultured vascular smooth muscle (VSM) cells pretreated with phorbol 12-myristate 13-acetate (PMA) and phorbol 12,13-butyrate (PDB) as PKC activator; 4$\alpha$-phorbol-didecanoate (PDD) as PKC non-activator. Nitrite production induced by IL-1$\beta$ was increased by the presence of increasing concentration of PMA ranging from 2 to 200 nM. However, in VSM cells pretreated with PMA and PDB, IL-1$\beta$-induced $NO_2$ production was decreased in proportion to the duration of pretreatment, and most significantly decreased in pretreatment time of 24 hours. Using RT-PCR method, the expression of iNOS mRNA induced by IL-1$\beta$ was decreased in VSM cells pretreated with PMA 200 nM for 24 hours. These results suggest that decrease in IL-I$\beta$-induced nitrite production by the pretreatment of PMA result from inhibition of iNOS expression and the inhibition related to PMA-induced PKC down-regulation.

Protein kinase C (PKC)가 interleukin-1 beta (IL-1$\beta$)에 의하여 산화질소(NO) 생성과정에 어떤 역할을 하는지를 검토하기 위하여, 혈관평활근세포에서 PKC 활성제인 phorbol 12-myristate 13-acetate (PMA)로 전처리한 후 IL-1$\beta$에 의하여 야기되는 NO생성을 nitrite ($NO_2$)로 정량하고, RT-PCR method를 이용하여 iNOS 발현에 미치는 영향을 검토하여 다음과 같은 결과를 얻었다. PMA (20, 200 nM)는 IL-1$\beta$에 의한$NO_2$ 생성을 유의하게 증가시켰다. PMA 200 nM, phorbol 12,13-dibutyrate 500 nM로 전처리하여 8, 24시간 노출된 세포에서 IL-1$\beta$에 의한 NO2생성이 현저히 감소되었으나, PKC 비활성제인 4$\alpha$-phorbol-didecanoate 200 nM로 전처리한 경우는 영향을 받지 아니하였다. PMA 농도를 달리하여 24시간 전처리한 경우 IL-1$\beta$에 의한 $NO_2$ 생성의 감소는 PMA의 농도가 20및 200 nM에서 현저하였다. RT-PCR method를 이용하여 iNOS 발현을 검토한바 IL-1$\beta$ 100U/ml에 의한 iNOS발현이 PMA전처리 및 cycloheximide 또는 actinomycin D존재로서 현저히 억제 되었다. 이상의 결과로 미루어 혈관평활근세포에서 PMA 전처리로 야기되는 IL-1$\beta$에 의한 NO 생성의 감소는, PKC 조절저하작용에 의한 iNOS 발현의 억제로 야기되는 것 같다.

Keywords

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