基质金属蛋白酶 8 抑制剂和洗必泰对 MDPC-23 细胞毒性、氧化应激和细胞因子水平的影响。

Effect of matrix metalloproteinase 8 inhibitor and chlorhexidine on the cytotoxicity, oxidative stress and cytokine level of MDPC-23.

机构信息

Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China.

出版信息

Dent Mater. 2018 Nov;34(11):e301-e308. doi: 10.1016/j.dental.2018.08.295. Epub 2018 Sep 17.

DOI:10.1016/j.dental.2018.08.295

PMID:30236973

Abstract

OBJECTIVE

This study investigated the effect of matrix metalloproteinase-8 inhibitor I (MMP8-I) and chlorhexidine (CHX) on the viability, oxidative stress and cytokine secretion of MDPC-23 under short-term (30min) and long-term (3 days) culture.

METHODS

MDPC-23 were treated with MMP8-I or CHX for 30min, 1day, 2days and 3days to detect the proliferation by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. In the following assays, MDPC-23 treated with 0.0003% CHX were referred to CHX group, treated with 8μM MMP8-I were MMP8-I group. Cells without additional treatment were regarded as control group. The cell cycle, reactive oxygen species (ROS) level, and apoptosis were assessed by flow cytometry. The cytokine level was quantified by enzyme-linked immunosorbent assay (ELISA).

RESULTS

In 30min, CHX at concentrations higher than 0.0003% dilution inhibited cell proliferation when compared to the control group. MMP8-I (0.1-500μM) showed no obvious cytotoxicity to MDPC-23, and MMP8-I (1000μM) inhibited cell proliferation. In 3 days, CHX (0.0003%) significantly inhibited cell growth, while MMP8-I (8μM) had no cytotoxicity. In the CHX group, the S phase population was decreased, and cellular ROS were increased in 3 days. In the MMP8-I group, the change of S phase population and cellular ROS was not significant compared with the control group. Cell apoptosis was not elevated in the MMP8-I group, while the apoptosis rate was increased in the CHX group both in 30min and 3 days. In 30min, CHX treatment significantly increased the secretion of interleukin (IL)-1β and IL-8, but slightly increased the secretion of IL-10, while MMP8-I caused no change in cytokines. In 3 days, CHX treatment significantly increased the secretion of IL-1β, IL-6, and IL-8, and inhibited the secretion of IL-10. MMP8-I treatment caused the increase of IL-6.

SIGNIFICANCE

Compared with CHX, MMP8-I at low concentration did not result in cytotoxicity, oxidative stress, or the disorder of immune response.

摘要

目的

本研究旨在探讨基质金属蛋白酶-8 抑制剂 I（MMP8-I）和洗必泰（CHX）对 MDPC-23 在短期（30 分钟）和长期（3 天）培养下的活力、氧化应激和细胞因子分泌的影响。

方法

用 MMP8-I 或 CHX 处理 MDPC-23 30 分钟、1 天、2 天和 3 天，通过 3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐（MTT）测定增殖。在以下实验中，用 0.0003% CHX 处理的 MDPC-23 被称为 CHX 组，用 8μM MMP8-I 处理的 MDPC-23 被称为 MMP8-I 组。未进行其他处理的细胞被视为对照组。通过流式细胞术评估细胞周期、活性氧（ROS）水平和细胞凋亡。通过酶联免疫吸附试验（ELISA）定量细胞因子水平。

结果

在 30 分钟时，与对照组相比，浓度高于 0.0003%稀释的 CHX 抑制细胞增殖。MMP8-I（0.1-500μM）对 MDPC-23 无明显细胞毒性，而 MMP8-I（1000μM）抑制细胞增殖。在 3 天时，CHX（0.0003%）显著抑制细胞生长，而 MMP8-I（8μM）无细胞毒性。在 CHX 组中，S 期细胞群减少，3 天时细胞内 ROS 增加。与对照组相比，MMP8-I 组 S 期细胞群和细胞内 ROS 的变化不明显。MMP8-I 组细胞凋亡率无升高，而 CHX 组 30 分钟和 3 天时细胞凋亡率均升高。在 30 分钟时，CHX 处理显著增加白细胞介素（IL）-1β和 IL-8 的分泌，但略微增加 IL-10 的分泌，而 MMP8-I 对细胞因子无影响。在 3 天时，CHX 处理显著增加 IL-1β、IL-6 和 IL-8 的分泌，并抑制 IL-10 的分泌。MMP8-I 处理导致 IL-6 的增加。