Observation of adequate stimulus methods for key initiative factor expression in human keratinocytes of anaphylactic disease
10.3969/j.issn.1001-1978.2015.04.027
- VernacularTitle:过敏性疾病关键启动因子在人角质形成细胞中表达的适宜刺激方法探索
- Author:
Hailiang LIU
;
Kaifan BAO
;
Xiaoyan JIANG
;
Xiao WEI
;
Xi YU
;
Yu TAO
;
Xiaoyu WANG
;
Yan WANG
;
Min HONG
- Publication Type:Journal Article
- Keywords:
HaCaT cells;
anaphylactic disease;
Poly (I:C);
TNF-α;
TSLP;
IL-33
- From:
Chinese Pharmacological Bulletin
2015;(4):576-581
- CountryChina
- Language:Chinese
-
Abstract:
Aim To explore the expressed level of ini-tiative key factors TSLP and IL-33 in a human kerati-nocyte cell line,HaCaT cells were chosen to be stimu-lated by different stimulants,and develop a stable and effective in vitro model to observe allergic sensitization. Methods HaCaT cells were cultured in K-SFM with different stimulants to screen out the stimulants which could significantly improve the expressed level of TSLP and IL-33.Expressed level of TSLP and IL-33 was an-alyzed by ELISA kits and immunofluorescence.Re-sults (1 )The dose-response relationship of single stimulant indicated that both Poly(I:C)and TNF-αcould significantly improve expressed level of TSLP and IL-33 in HaCaT cells,but the rest of stimulants was not observed significant stimulation in concentration range of this experiment.(2)Dose-effect relationship of combined stimulants indicated that poly(I ∶C)1 00 mg·L -1 combined with TNF-α20 μg·L -1 was the most efficient.(3)Time-effect relationship of the a-bove-mentioned combined stimulants showed that 1 2 h was the optimal time of stimulation.Conclusions Different stimulants and different time result in various expressed levels of TSLP and IL-33 in HaCaT cells.1 2 h stimulus duration of Poly(I:C)1 00 mg·L -1 com-bined with TNF-α20 μg · L -1 is the most efficient stimulating way.This result provides an effective in vitro model to study the pathomechanism and drug effi-cacy of allergic sensitization.
