PCF regulating iNOS / NO and HSP inhibits UVB-induced apoptosis of HaCaT cells
|Keywords||PCF (PCF) Ultraviolet B (UVB) Nitric oxide synthase (iNOS) / nitric oxide (NO) Heat shock protein (HSP) Cyclooxygenase -2 (COX-2)|
The purpose of the establishment of iNOS transfected HaCaT cell model , the use of UVB-induced HaCaT cell apoptosis model , inducible nitric oxide synthase ( iNOS ) , nitric oxide ( NO ) , heat shock protein (HSP) and cyclooxygenase - 2 (COX-2) point of view , to study the the PCF (Polypeptide from Chlamys forreri, PCF) inhibit UVB - induced apoptosis of HaCaT cells molecular mechanism . Established iNOS stably transfected transiently transfected HaCaT cell model . Copy of 20 mJ / cm UVB -induced apoptosis of HaCaT cells model . The experimental design is divided into six groups: control group , the UVB model group , UVB iNOS - specific inhibitor SMT group , UVB 5.69 mmol / L PCF group , UVB 2.84mmol / L PCF group , UVB 1.42 mmol / LPCF group . Determination of the rate of apoptosis ; Hoechst 33258 fluorescence staining cells after UVB irradiation, electron spin resonance (ESR) detection of NO generated by the impact of change and its PCF ; iNOS mRNA expression by RT- PCR detection ; Protein Western blotting of iNOS , HSP27 , HSP70 , HSP90 and COX -2 protein expression . 20 mJ / cm ~ 2 UVB -induced the HaCaT cell iNOS mRNA and protein expression increased ( P <0.05 ) , and thus NO release increased significantly ( P <0.01 ) , and UVB radiation damage feedback upregulation of HSP70, HSP90 expression (P <0.05). INOS gene transfection HaCaT cells , NO release of a large number increased (P <0.05). 1.42 ~~ 5.69 mmol / L dose range PCF 1 mmol / L SMT significantly inhibited UVB - induced HaCaT cell apoptosis ( P <0.05 ) ; PCF within the dose range of 1.42 - 5.69mmol / L inhibited 20 mJ / cm ~ 2 UVB - induced HaCaT cell iNOS expression the ANO release (P <0.05, P <0.01), and raised the protective protein HSP70 expression of HSP90 (P <0.05); higher after transfection of iNOS expression in HaCaT cells , the PCF can inhibit the release of NO ( P <0.01 ) . Conclusion PCF could inhibit 20 mJ / cm to 2 UVB -induced apoptosis of HaCaT cells , its role in the mechanism of inhibition of UVB-induced iNOS activation , NO release increased , and upward of HSP70 of HSP90 -related .