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白细胞介素1#beta#抑制心肌成纤维细胞胶原合成并促进其分解

肖华, 季爱民, 李志粱, 宋旭东, 苏丹, 陈爱华

南方医科大学珠江医院心内科.广东,广州 510282

摘要

该文应用~(3)H--胸腺嘧啶核苷(~(3)H--thymidine, ~(3)H--TdR)掺入法及~(3)H--脯氨酸(~(3)H--proline, ~(3)H--Pro)掺入法观察白细胞介素1#beta#(interleukin--1#beta#, IL--1#beta#)对Spague--Dawley乳鼠心肌成纤维细胞DNA及胶原合成的影响，并用明胶酶谱法和Western blot检测基质金属蛋白酶(matrix metalloproteinases, MMPs) MMP--2、MMP--9活性及MMP--2和MMP--9蛋白表达，用RT--PCR检测MMP--2、MMP--9的mRNA表达。结果显示：(1)不同浓度的IL--1#beta# (0.1、1、10、100 ng/mL)作用于细胞24 h后，各组~(3)H--TdR掺入量明显较对照组低，同时~(3)H--Pro掺入量明显降低；而0.01 ng/mL的IL--1#beta#作用于CFs后，对~(3)H--TdR掺入量和~(3)H--Pro掺入量无明显影响；(2)不同剂量的IL--1#beta#均刺激MMP--2及MMP--9的活性升高，并呈剂量依赖性。IL--1#beta# 增加 MMP--2蛋白表达，分别为(0.01 ng/L：1.35±0.05倍；0.1 ng/mL：1.56±0.10倍；{sl P}<0.05；1 ng/m：2.15±0.32倍；10 ng/mL：2.34±0.19倍; 100 ng/mL：2.41±0.26倍；{sl P}<0.01)，同时增加MMP--9 蛋白表达(0.01 ng/mL：1.38±0.20倍；{sl P}<0.05；0.1 ng/mL：2.37±0.21倍；1 ng/mL：2.56±0.31倍；10 ng/mL：2.60±0.19倍；100 ng/mL：2.67±0.15倍；{sl P}<0.01)；(3) MMP--2和MMP--9的活性、mRNA和蛋白表达均较对照组增高。IL--1#beta#刺激MMP--2 mRNA表达升高(0.01 ng/mL：1.81±0.16倍；{sl P}<0.5；0.1 ng/mL：2.17±0.19倍；1 ng/mL：2.18±0.04倍；10 ng/mL：2.27±0.06倍；100 ng/mL：2.31±0.07倍；{sl P}<0.01)及MMP--9 mRNA 表达升高(0.01 ng/mL：1.36±0.13倍；{sl P}<0.05；0.1 ng/mL：1.49±0.08倍；1 ng/mL：1.50±0.11倍；10 ng/mL： 1.51±0.24倍；100 ng/mL：1.52±0.31倍；{sl P}<0.01)。以上结果表明，IL--1#beta#通过减少心肌成纤维细胞的细胞分裂来降低胶原的合成，同时促进MMP--2和MMP--9的转录及转录后的表达来促进胶原的分解，提示其在心肌重塑过程中起一定作用。

关键词： 白细胞介素1#beta#; 成纤维细胞; 胶原; 基质金属蛋白酶

Interleukin--1#beta# inhibits collagen synthesis and promotes its decomposition in cultured cardiac fibroblasts

Xiao Hua, Ji Aimin, Li Zhiliang, Song Xudong, Su Dan, Chen Aihua

Department of Cardiology,the Zhujiang Hospital,Nanfang Medical University.Guangzhou 510282,Guangdong

Abstract

The present study aimed to investigate the effects of intereukin-1#beta# at different doses on collagen synthesis and decomposition in cultured cardiac fibroblasts in neonatal Sprague-Dawley rat. Cardiac fibroblasts were treated with IL-1#beta# (0.01, 0.1, 1, 10, 100 ng/mL）for 24 h. Cell DNA synthesis was measured by ~(3)H-thymidine (~(3)H-TdR) incorporation and collagen synthesis was measured by ~(3)H-proline (~(3)H-Pro) incorporation. Matrix metalloproteinase (MMP) activity was measured by gelatinase zymography. MMP-2, MMP-9 protein expressions were measured by Western blot. mRNA expressions of MMP-2 and MMP-9 were detected by reversed transcription-ploymerase chain reaction (RT-PCR). Compared with that in the control group, the incorporation of ~(3)H-TdR and ~(3)H-Pro decreased in high-dose IL-1#beta# groups (≥0.1 ng/mL) but not in low-dose IL-1#beta# group (0.01 ng/mL). IL-1#beta# significantly increased MMP-2 and MMP-9 activities. IL-1#beta# also dose-dependently increased the protein expressions of MMP-2 (fold change; 0.01 ng/mL: 1.35±0.05; 0.1 ng/mL: 1.56±0.10; P<0.05; 1 ng/mL: 2.15±0.32; 10 ng/mL: 2.34±0.19; 100 ng/mL: 2.41±0.26; P<0.01) and MMP-9 (fold change; 0.01 ng/mL: 1.38±0.20, P<0.05; 0.1 ng/mL: 2.37±0.21; 1 ng/mL: 2.56±0.31; 10 ng/mL: 2.60±0.19; 100 ng/mL: 2.67±0.15; P<0.01), respectively. Similar results were observed in mRNA expression of MMP-2 (fold change; 0.01 ng/mL: 1.81±0.16, P<0.05; 0.1 ng/mL: 2.17±0.19; 1 ng/mL: 2.18±0.04; 10 ng/mL: 2.27±0.06; 100 ng/mL: 2.31±0.07; P<0.01) and MMP-9 (fold change; 0.01 ng/mL: 1.36±.13; P<0.05; 0.1 ng/mL: 1.49±0.08; 1 ng/mL: 1.50±0.11; 10 ng/mL: 1.51±0.24; 100 ng/mL: 1.52±0.31; P<0.01). These results suggest that IL-1#beta# decreases collagen synthesis and activates MMPs through transcriptional and posttranslational processes via degrading collagen in a dose-dependent way. Elevation of IL-1#beta# after myocardial infarction is possibly involved in the process of post-myocardial infarction ventricle remodeling, and the concentration of IL-1#beta# is possibly a major factor which affects the degree of ventricle remodeling.

Key words: interleukin-1#beta#;Fibroblasts;collagen;matrix metalloproteinases

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引用本文：

肖华, 季爱民, 李志粱, 宋旭东, 苏丹, 陈爱华. 白细胞介素1#beta#抑制心肌成纤维细胞胶原合成并促进其分解[J]. 生理学报 2008; 60 (3): 355-361.

Xiao Hua, Ji Aimin, Li Zhiliang, Song Xudong, Su Dan, Chen Aihua. Interleukin--1#beta# inhibits collagen synthesis and promotes its decomposition in cultured cardiac fibroblasts. Acta Physiol Sin 2008; 60 (3): 355-361 (in Chinese with English abstract).