CAJ | 학술논문

目的：研究不同刺激条件对人角质形成细胞 HaCaT细胞中 TSLP、IL-33表达水平的影响，探讨过敏性疾病中关键启动因子 TSLP、IL-33体外表达细胞模型的最佳刺激方法。方法应用角质形成细胞无血清培养液（K-SFM）体外培养 HaCaT 细胞，给予不同刺激剂，筛选出明显促进 HaCaT细胞中 TSLP 和 IL-33表达的刺激剂。进而考察单独与联合刺激剂时的量效关系，最后对选出的刺激剂进行时效关系考察。TSLP 和 IL-33表达水平采用 ELISA 和免疫荧光法检测。结果（1）Poly（I：C）与 TNF-α两种刺激剂单独使用时均能明显刺激 HaCaT 细胞分泌 TSLP 和 IL-33，其余刺激剂在本实验浓度范围内未见明显差异。（2）Poly（I：C）100 mg· L －1与 TNF-α20μg·L －1联合刺激对 HaCaT 细胞表达 TSLP和 IL-33的促进作用最为明显。（3）对 Poly（I：C）100 mg· L －1与 TNF-α20μg·L －1联合刺激 HaCaT 细胞的时效关系考察发现，刺激12 h HaCaT 细胞中 TSLP 和 IL-33的表达水平最高。结论不同刺激剂和刺激时间对体外刺激 HaCaT细胞表达细胞因子 TSLP 和 IL-33的效应不同，其中以 Poly （I：C）100 mg·L －1与 TNF-α20μg·L －1联合刺激 HaCaT 细胞12 h 后，TSLP 和 IL-33的表达水平升高最为明显。该结果为过敏性疾病的病理机制及药物作用研究提供了合适的方法。
목적：연구불동자격조건대인각질형성세포 HaCaT세포중 TSLP、IL-33표체수평적영향，탐토과민성질병중관건계동인자 TSLP、IL-33체외표체세포모형적최가자격방법。방법응용각질형성세포무혈청배양액（K-SFM）체외배양 HaCaT 세포，급여불동자격제，사선출명현촉진 HaCaT세포중 TSLP 화 IL-33표체적자격제。진이고찰단독여연합자격제시적량효관계，최후대선출적자격제진행시효관계고찰。TSLP 화 IL-33표체수평채용 ELISA 화면역형광법검측。결과（1）Poly（I：C）여 TNF-α량충자격제단독사용시균능명현자격 HaCaT 세포분비 TSLP 화 IL-33，기여자격제재본실험농도범위내미견명현차이。（2）Poly（I：C）100 mg· L －1여 TNF-α20μg·L －1연합자격대 HaCaT 세포표체 TSLP화 IL-33적촉진작용최위명현。（3）대 Poly（I：C）100 mg· L －1여 TNF-α20μg·L －1연합자격 HaCaT 세포적시효관계고찰발현，자격12 h HaCaT 세포중 TSLP 화 IL-33적표체수평최고。결론불동자격제화자격시간대체외자격 HaCaT세포표체세포인자 TSLP 화 IL-33적효응불동，기중이 Poly （I：C）100 mg·L －1여 TNF-α20μg·L －1연합자격 HaCaT 세포12 h 후，TSLP 화 IL-33적표체수평승고최위명현。해결과위과민성질병적병리궤제급약물작용연구제공료합괄적방법。
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.