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신경교세포 및 RAW 264.7 세포에서 Protein kinase의 활성에 의한 유도성 Nitric oxide synthase의 발현
박상철,노삼길,배소현,박지선,이충재,허강민,석정호,이재흔 충남대학교 의학연구소 2003 충남의대잡지 Vol.30 No.2
NO(nitric oxide) plays an important role as neurotransmitter or cytokine, and pathologic factor for some diseases by the large amount production with iNOS(inducible NO synthase) expression in macrophages or glial cells. The expression of iNOS is regulated by various cytokines, protein kinases and transcription factors. In this experiment, to investigate the roles of progein kinase and NF-kB for iNOS expression, the effects of PMA(phorbol 12-myristate 13-acetate), cAMP, and various protein kinase inhibitors on LPS(lipopolysaccharide)-induced iNOS mRNAN expression and nuclear NF-kB binding complex were examined in C6 glial cells and RAW 264.7 cells. In C6 glial cells, iNOS mRNA expression by LPS was induced from 1 hour and peak at 3 hour after treatment. In RAW 264.7 cells, the mRNA was observed from 3 hour and peak at 6 hour. PMA enhanced markedly LPS-induced iNOS mRNA expression and NF-kB binding complex in C6 glial cells, but did not much influence on LPS-induced iNOS mRNA expression in RAW 264.7 cells, in spite of increased LPS-induced NF-kB binding complex at 30 min. cAMP(dibutyryl cAMP) did not much influence on LPS-induced iNOS mRNA expression, by increased LPS-induced NF-kB binding complex in C6 glial cells. However, in RAW 264.7 cells, cAMP increased slightly LPS-induced iNOS mRNA expression without change of NF-kB binding complex. Staurosporine did not influence on LPS-induced iNOS mRNA expression and NF-kB binding complex in C6 glial cells, but in RAW 264.7 cells, decreased LPS-induced iNOS mRNA expression and NF-kB binding complex. Ro-31-8220 did not much influence on LPS-induced iNOS mRNA expression and NF-kB binding complex in C6 glial cells, but in RAW 264.7 cells, decreased significantly LPS-induced iNOS mRNA expression in spite of increased LPS-induced NF-kB binding complex for 3hours. G 6976 did not much influence on LPS-induced iNOS mRNA expression with decreased NF-kB binding complex in C6 glial cells, but in RAW 264.7 cells, decreased iNOS mRNA expression without influence on LPS-induced NF-kB binding complex. Genistein did not influence on LPS-induced iNOS mRNA expression and NF-kB binding complex in C6 glial cells, but in RAW 264.7 cells, decreased LPS-induced iNOS mRNA expression inspite of increased NF-kB binding complex. These results suggest that LPS-induced regulation of iNOS expression or NF-kB activity in C6 glial cells, might be different from RAW 264.7 cells through various protein kinases or other factors.
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일차배양된 설치류 호흡기 상피세포로부터의 점액소 분비에 대한 수종 약물의 영향
이충재,석정호,이재흔,허강민,박지선,배소현,노삼길,박상철 충남대학교 의학연구소 2003 충남의대잡지 Vol.30 No.2
1. PKC activator인 PMA는 일차배양 HTSE세포로부터의 뮤신분비를 0.1μM 농도에서 30%, 1μM 농도에서 80% 가량 증가시켰다. 2. 식물 유래 성분으로, flavonoid의 일종인 TFR은 일치배양 HTSE 세포로부터의 뮤신분비를 10μM 농도에서 50%, 100μM 농도에서 80% 가량 증가시켰다. 3. 양이온성 폴리펩티드인 PLL 및 PLA는 일차 배양 HTSE 세포로부터의 뮤신분비를 0.01 - 10μM 농도에서 용량의존적으로 감소시켰다. 4. 결론적으로, 본 연구에서 얻어진 결과들은 새로운 거담제 및 점액용해제나 단백분해 효소제가 아닌 호흡기 류신의 생성/분비를 조절해 줄 수 있는 신개념의 약물을 개발함에 있어 극히 일부분이나마 단서를 제공하고 있다고 하루 수 있을 것이다. In the present study, we tried to investigate whether phorbol myristate acetate(PMA), trihydroxymethoxy-flavanone rutinoside(TFR) and cationic polypeptides significantly affect mucin release(secretion) from cultured hamster tracheal surface epithelial cells. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24 hr and chased for 30 min in the presence of each agent to assess the effect on 3H-mucin release. The results were as follows : (1) Both PMA and TFR significantly increased mucin release from cultured HTSE cells ; (2) Cationic polypeptides including po1y-L-lysine(PLL, mw 7,500) and poly-L-arginine(PLA, mw 10,800) significantly inhibited mucin release from cultured HTSE cells, in a dose-dependent manner. This finding suggests us that PMA and TFR be further studied for the possible use as mild expectorants and cationic polypeptides might function as a regulator for hyper-secretion of mucus, both by direct acting on airway mucin-secreting cells, during the treatment of chronic airway diseases.
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