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사람의 복막중피세포에서 Interleukin-1β 자극에 의한 Fibronectin 생성의 세포내 신호전달에 있어 Atypical Protein Kinase C의 역할
양원석 ( Yang Won Seog ),김병식 ( Kim Byeong Sig ),김순배 ( Kim Sun Bae ),박수길 ( Park Su Gil ),박정식 ( Park Jeong Sig ) 대한신장학회 2003 Kidney Research and Clinical Practice Vol.22 No.4
배 경 : Protein kinase C (PKC)에는 diacylglycerol (DAG)에 의해 활성화되는 conventional type (PKC-α, -βⅠ, -βⅡ, -γ )과 novel type (PKC-δ, -ε, -θ, -η)외에 DAG에 의해 활성화되지 않는 atypical type (PKC-ζ, -ι)이 있다. Glucose 자극에 의한 fibronectin 생성 과정에 관여하는 PKC isoenzyme은 DAG에 의해 활성화되는 PKC로 알려져 있다. 본 실험에서는 사람의 복막중피세포에서 IL-1β자극에 의한 fibronectin 생성의 신호전달 과정에 PKC가 관여하는 지와 그 isoenzyme에 대해 알아보고자 하였다. 방 법 : Fibronectin mRNA양은 northern blot assay로 측정하였고, 세포내 phosphorylated PKC ζ/ι 단백질 양은 western blot으로 비교하였다. 결 과 : 복막중피세포를 PKC억제제인 calphostin C (500, 750, 1000 nM)로 전처치 한 다음 IL-1β (1 ng/mL)로 자극하였을 때 fibronectin mRNA 발현은 calphostin C 용량에 비례하여 억제되었다. 다른 PKC억제제인 GF109203X (1, 5, 10 μM)도 IL-1β 자극에 의한 fibronectin mRNA 발현을 용량에 비례하여 억제시켰다. 반면에, conventional 및 novel PKC 자극제인 phorbol 12-myristate 13-acetate (PMA)는 fibronectin mRNA 발현을 증가시켰다. PMA를 72시간 작용시켜 PKC를 소모시킨 상태에서 PMA를 다시 투여해도 fibronectin mRNA 발현이 자극되지 않는 반면, IL-1β(1 ng/mL)는 fibronectin mRNA 발현을 자극하였다. Myristoylated PKC ζ/ι pseudosubstrate (5, 10, 15, 20 μM)는 IL-1β자극에 의한 fibronectin mRNA 발현을 용량에 비례하여 억제시켰고, 20 μM에서는 IL-1β자극에 의한 fibronectin mRNA 발현을 완전히 억제하였다. 대조군으로 투여한 myristoylated PKC [19-27] pseudosubstrate (20 μM)는 IL-1β자극에 의한 fibronectin mRNA 발현에 영향을 주지 않았다. IL-1β 자극에 의해 PKC ζ/ι가 활성화되는 지를 보기 위해 시행한 western blot에서는 IL-1β자극에 의해 PKC ζ/ι의 활성화된 형태인 phosphorylated PKC ζ/ι가 증가하였다. 결 론 : 이상의 결과는 복막중피세포에서 IL-1β에 의한 fibronectin 생성이 atypical PKC인 PKC ζ/ι 활성화를 통해 일어남을 시사한다. Background : Protein kinase C (PKC)s consist of three groups of isoenzyme; conventional, novel and atypical PKCs. Diacylglycerol (DAG) activates both conventional and novel PKCs, but not atypical PKCs. High glucose-induced fibronection production was shown to be mediated by activation of DAG-sensitive PKCs. In this study, we investigated whether PKC mediates IL-1β-induced fibronectin mRNA expression, and the subtypes of PKC involved in the process. Methods : Fibronection mRNA level and phosphorylated PKC ζ/ι in total cell lysate were measured by Northern blot and Western blot, respectively. Results : Pretreatment of HPMCs with calphostin C, a pan-PKC inhibitor, at doses of 500, 750 and 1,000 nM caused dose-dependent inhibition of IL-1β (1 ng.mL)-induced fibronection mRNA level. GF109203X, another pan-PKC inhibitor, at doses of 1, 5 and 10 ㎛ also downregulated IL-1β (1 ng/mL)-induced fibornectin mRNA level in a dose-de-pendent manner. Phorbol 12-myristate 13-acetate(PMA), an activator of conventional and novel PKCs, stimulated fibornectin mRNA level at doses of 1, 10 and 100nM. After prolonged treatment of the cells for 72 hr with PMA, another dose of PMA did not increase fibronectin mRNA level, while IL-1β (1 ng/mL) still stimulated it. Pretreatment of the cells with 5, 10, 15 and 20 ㎛ of myristoylated PKC ζ/ι pseudosubstrate inhibited IL-1β (1 ng/mL)-in-duced fibronectin mRNA level in a dose-dependent manner, while 20 ㎛ of myristoylated PKC [19-27] pseudosubstrate, given as a control, had no effect. Stimulation of fibronectin mRNA level by IL-1β (1 ng/mL) was completely prevented by 20 ㎛of myristoylated PKC ζ/ιpseudosubstrate. IL-1β (1 ng/mL) increased phosphorylated PKC ζ/ι, an active form of the enzyme. Conclusion : IL-1β-induced fibronectin production in HPMCs occurs by way of activation of atypical PKCs (PKC ζ/ι). (Korean J Nephrol 2003;22(4):340-348)
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골프장의 마케팅믹스변인과 브랜드자산 및 행동의도의 구조적 관계
박정환 ( Joung-hwan Park ),김병식 ( Byeong-sig Kim ) 한국체육대학교 체육과학연구소 2021 스포츠사이언스 Vol.39 No.3
The purpose of this study was to identify structural relationships between golf course marketing mix variables, brand assets, and behavioral intentions among golf course users. The subjects applied the convenience sampling method and snowball sampling to 250 golfers using golf courses in Seoul and Gyeonggi Province, and finally 209(90.0%) out of 150 golf course users and 100 online questionnaires were analyzed. The collected data were obtained using the SPSSWIN 21.0 and AMOS 21.0 programs to perform descriptive statistics, reliability analysis, confirmative factor analysis, correlation analysis, and structural equation models. First, in terms of the influence of golf course marketing mix variables on brand assets, location and price affected perceived quality, and promotion, location, and price affected brand recognition and brand image. There was no casual link other than that. Second, golf course marketing mix variables and brand assets were found to have no effect on behavioral intention. Golf courses should establish and implement marketing strategies to build brand assets of golf courses from a macro perspective so that they can respond well to the situation after COVID19.
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