Effects of L-carnitine, erythritol and betaine on pro-inflammatory markers in primary human corneal epithelial cells exposed to hyperosmotic stress.
Effects of L-carnitine, erythritol and betaine on pro-inflammatory markers in primary human corneal epithelial cells exposed to hyperosmotic stress.
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DOI：
10.3109/02713683.2014.957776
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发表时间：
2015-07
影响因子：
2
通讯作者：
Pflugfelder SC
中科院分区：
文献类型：
--
作者：
Hua X;Su Z;Deng R;Lin J;Li DQ;Pflugfelder SC
探讨渗透保护剂对高渗应激下原代人角膜上皮细胞（HCECs）促炎介质产生的影响。在等渗培养基（312mOsM）中培养的HCECs在2-20mM左旋肉碱、赤四糖醇或甜菜碱的孵育下切换到高渗培养基，孵育不同时间。采用RT-qPCR和ELISA检测HCECs中促炎标志物的mRNA表达和蛋白生成情况。高渗介质以渗透压依赖的方式显著刺激HCECs中促炎细胞因子TNF-α、IL-1β和IL-6以及趋化因子IL-8、CCL2和CCL20的mRNA和蛋白表达。左旋肉碱、赤藓糖醇和甜菜碱均能显著抑制这些促炎介质的表达。l -肉碱表现出最大的抑制作用，使暴露于450mOsM的HCECs中TNF-α（从12.3倍降至5.7倍）、IL-1β（从2.2倍降至0.9倍）、IL-6（从7.3倍降至2.9倍）、IL-8（从4.6倍降至2.0倍）、CCL2（从15.3倍降至3.5倍）和CCL20（从4.1倍降至1.5倍）的mRNA水平降低54-77%。左旋肉碱、赤藓糖醇和甜菜碱也显著抑制了受刺激的TNF-α、IL-1β、IL-6和IL-8蛋白的产生。l -肉碱抑制了暴露于450mOsM的HCECs的TNF-α（从81.3降至17.4pg/ml）、IL-1β（从56.9降至29.2pg/ml）、IL-6（从12.8降至4.6ng/ml）和IL-8（从21.2降至10.9ng/ml）的49-79%的受刺激蛋白水平。有趣的是，高渗透压刺激TNF-α、IL-1β和IL-6 mRNA和蛋白水平的升高被TRPV1激活抑制剂capsazepine显著抑制。l -肉碱、赤四糖醇和甜菜碱作为渗透保护剂，通过TRPV1途径抑制高渗应激下HCECs的炎症反应。渗透保护剂可能对减少干眼病的先天性炎症有疗效。
To explore the effects of osmoprotectants on pro-inflammatory mediator production in primary human corneal epithelial cells (HCECs) exposed to hyperosmotic stress. HCECs cultured in iso-osmolar medium (312mOsM) were switched to hyperosmotic media with or without prior incubation with 2–20mM of L-carnitine, erythritol or betaine for different time periods. The mRNA expression and protein production of pro-inflammatory markers in HCECs were evaluated by RT-qPCR and ELISA. Hyperosmolar media significantly stimulated the mRNA and protein expression of pro-inflammatory cytokines, TNF-α, IL-1β and IL-6, and chemokines, IL-8, CCL2 and CCL20 in HCECs in an osmolarity dependent manner. The stimulated expression of these pro-inflammatory mediators was significantly but differentially suppressed by L-carnitine, erythritol or betaine. L-carnitine displayed the greatest inhibitory effects and down-regulated 54–77% of the stimulated mRNA levels of TNF-α (down from 12.3 to 5.7 fold), IL-1β (2.2 to 0.9 fold), IL-6 (7.3 to 2.9 fold), IL-8 (4.6 to 2.0 fold), CCL2 (15.3 to 3.5 fold) and CCL20 (4.1 to 1.5 fold) in HCECs exposed to 450mOsM. The stimulated protein production of TNF-α, IL-1β, IL-6 and IL-8 was also significantly suppressed by L-carnitine, erythritol and betaine. L-carnitine suppressed 49–79% of the stimulated protein levels of TNF-α (down from 81.3 to 17.4pg/ml), IL-1β (56.9 to 29.2pg/ml), IL-6 (12.8 to 4.6ng/ml), and IL-8 (21.2 to 10.9ng/ml) by HCECs exposed to 450mOsM. Interestingly, hyperosmolarity stimulated increase in mRNA and protein levels of TNF-α, IL-1β and IL-6 were significantly suppressed by a TRPV1 activation inhibitor capsazepine. L-carnitine, erythritol and betaine function as osmoprotectants to suppress inflammatory responses via TRPV1 pathway in HCECs exposed to hyperosmotic stress. Osmoprotectants may have efficacy in reducing innate inflammation in dry eye disease.