붉은자루동충하초의 항염물질의 생리활성확인(1)

나. 붉은자루동충하초의 메탄올 추출물의 PGE2 및 COX-2 단백질 발현억제효과

PGE2는 활성화 된 대식세포의 pro-inflammation enzyme인 cyclooxygenase에 의해서 생성되어지는 이때 대식세포를 LPS로 활성화시킨 후에 세포배양에서 PGE2양을 kit로 이용하여 측정한 결과 CPME의 농도에 따라서 감소하는 것을 알 수가 있었으며, Western blot과 RT-PCR을 통해서 이 물질의 생성의 감소는 CPME의 농도에 따라서 합성 및 전사가 억제되는 것을 알 수강 있었다(Fig 22).

 

Fig. 22. Effects of the CPME on PGE2 production and COX-2 gene ｅxpression.

RAW264.7 cells were treated with LPS (1 ㎍/ml) in the presence or absence of the CPME. (A) PGE2 concentration was measured in the 16 h culture media using a commercial ELISA kit. Data are shown the mean ± SD (n=4). ＊, p＜0.05; ＊＊, p＜0.01 versus LPS alone. (B) COX-2 protein levels were determined in cells stimulated with LPS for 12 h by Western blot analysis. The membrane was rehybridized with antibody against β-actin to verify equal loading of protein in each lane. (C) COX-2 mRNA levels were determined in cells stimulated with LPS for 8 h by RT-PCR analysis. β-actin was used as an internal control.