Cong Lin, Xia Yi-Ping, Zhao Gui-Qiu, Lin Jing, Xu Qiang, Hu Li-Ting, Qu Jian-Qiu, Peng Xu-Dong
Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266000, Shandong Province, China.
Int J Ophthalmol. 2015 Oct 18;8(5):866-71. doi: 10.3980/j.issn.2222-3959.2015.05.03. eCollection 2015.
To observe the expression of vitamin D receptor (VDR) in human specimen and immortalized human corneal epithelium cells (HCEC) when challenged with fusarium solani. Moreover, we decided to discover the pathway of VDR expression. Also, we would like to detect the expression of cathelicidin antimicrobial peptide (CAMP) in the downstream pathway of VDR.
Immunohistochemistry was used to examine the VDR expression in HCEC from healthy and fungal keratitis patients. Real time quantitative polymerase chain reaction (qPCR) was performed to observe the messenger ribonucleic acid (mRNA) change of VDR when immortalized HCEC were challenged with fusarium solani for different hours. CAMP was detected at both mRNA and protein levels.
We found out that the VDR expression in fusarium solani keratitis patients' specimen was much more than that in healthy people. The mRNA and protein expression of VDR increased when we stimulated HCEC with fusarium solani antigen (P<0.01) and it could be inhibited by toll like receptor 2 (TLR2) monoclonal antibody. The CAMP expression was decreased because of fusarium solani antigen stimulation (P<0.01).
The VDR expression can be increased via TLR2/1-VDR pathway while the CAMP expression is decreased by the stimulation of fusarium solani antigen.
观察人标本及永生化人角膜上皮细胞(HCEC)在茄病镰刀菌刺激下维生素D受体(VDR)的表达。此外,我们决定探寻VDR表达的途径。同时,我们希望检测VDR下游途径中cathelicidin抗菌肽(CAMP)的表达。
采用免疫组织化学法检测健康人和真菌性角膜炎患者HCEC中VDR的表达。对永生化HCEC用茄病镰刀菌刺激不同时间后,采用实时定量聚合酶链反应(qPCR)观察VDR信使核糖核酸(mRNA)的变化。从mRNA和蛋白质水平检测CAMP。
我们发现茄病镰刀菌角膜炎患者标本中VDR的表达远高于健康人。用茄病镰刀菌抗原刺激HCEC时,VDR的mRNA和蛋白质表达增加(P<0.01),且可被Toll样受体2(TLR2)单克隆抗体抑制。茄病镰刀菌抗原刺激导致CAMP表达降低(P<0.01)。
VDR表达可通过TLR2/1-VDR途径增加,而茄病镰刀菌抗原刺激可使CAMP表达降低。