간세포성장인자가 메산지움 세포의 TGF-β, Fibronectin, Collagen Ia 생성에 미치는 영향 = Effect of HGF on the Production of TGF-β, Fibronectin, and Collagen Ia in the Murine Mesangial Cells

다국어 초록 (Multilingual Abstract)

Background:Hepatocyte growth factor (HGF) is a multifunctional protein especially implicated in tissue repair and regeneration. In a murine model of chronic glomerulonepritis, subcutaneous injection of recombinant HGF inhibited renal fibrosis. Although these favorable results were accompanied by decreased expression of TGF-β in renal tissue, the action of HGF on TGF-β remains unclear. We studied whether HGF has an inhibitory effect on TGF-β production in vitro.
Methods:Mouse mesangial cells grown in 5 mM glucose containing DMEM (NG, normal glucose) were exposed with 25 mM glucose medium (HG, high glucose) alone or in the presence of recombinant HGF (10 ng/mL) for 48 hours. TGF-β in culture supernatants were measured by sandwich enzyme-linked immunosorbent assay. Secretion of fibronectin and collagen Ia was measured by western blotting and immunoprecipitaiton. TGF-β, fibronectin and collagen Ia mRNA levels were measured by real-time PCR and internally corrected with GAPDH.
Results:High glucose significantly increased TGF-β secretion (p<0.05), but HGF could not prevent high glucose-induced TGF-β release. High glucose increased the secretion of both fibronectin and collagen Ia, which was blocked by HGF (p<0.05).
Conclusion:In cultured murine mesangial cells HGF did not have effect on TGF-β production, but inhibited the secretion of both fibronectin and collagen Ia. These results suggest that HGF may have indirect inhibitory effect on the action of TGF-β.

국문 초록 (Abstract)

목 적:세포분열 촉진, 운동성 증가, 형태형성 촉진과 자멸사 억제 및 세포재생 등 다양한 생물학적 활성을 가지는 간세포성장인자 (Hepatocyte growth factor, HGF)는 만성사구체신염 동물모델에서 섬유화 억제 작용이 있으며, 이는 Transforming growth factor-β (TGF-β)의 신조직내 발현 감소와 관련이 있다고 추정된다. 세포외 기질 축적과 신조직의 섬유화과정에서 중요한 TGF-β에 대한 HGF의 효과를 알기 위해 본 연구를 시행하였다.
방 법:정상농도 포도당 배양액에 배양한 생쥐 메산지움 세포를 고농도 포도당 배양액에 노출한 후 유전자 재조합 HGF (10 ng/mL)를 배양액에 첨가하여 TGF-β 및 세포외 기질을 형성하는 fibronectin 및 collagen Ia 생성을 측정하였다. TGF-β 분비의 변화는 ELISA로, fibronectin 및 collagen Ia 생성의 변화는 Western blot과 면역침강법을 이용하여 측정하였다. TGF-β, fibronectin 및 collagen Ia mRNA의 발현 변화도 측정하였다.
결 과:고농도 포도당 배양액에서 배양한 경우 정상농도 포도당 배양액에 비해 48시간 후 TGF-β 분비량이 증가하였으나 (p<0.05), 유전자 재조합 HGF 첨가에 의해 고농도 포도당으로 증가된 TGF-β 분비는 억제되지 않았다. 고농도 포도당 배양액 처리시 정상농도 포도당 배양액에 비해 fibronectin 및 collagen Ia 생성이 증가되었으며, 유전자 재조합 HGF 첨가 후 fibronectin 및 collagen Ia 생성이 감소하였다 (p<0.05).

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