摘要
目的:通过观察川芎嗪(tetramethylpyrazine,TMP)对血管紧张素Ⅱ(angiotensin Ⅱ,AngⅡ)诱导的大鼠心肌成纤维细胞(cardiac fibroblast,CFB)增殖及Ⅰ型胶原合成的影响,探讨其抗心肌纤维化的作用机制。方法:差速贴壁法提取原代SD乳鼠CFB,采用3～4代CFB进行检测。以甲基噻唑基四唑(methyl thiazolyltetrazolium,MTT)法测定细胞数目,观察CFB增殖情况;收集细胞培养上清,以酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测Ⅰ型胶原的合成;提取细胞总mRNA,以逆转录聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)半定量检测Ⅰ型胶原mRNA的表达情况。结果:(1)MTT结果显示,0.1μmol/LAngⅡ组CFB光密度(optical density,OD)值高于空白对照组(P<0.05)。0.1μmol/LAngⅡ+800μg/mLTMP组和0.1μmol/LAngⅡ+600μg/mLTMP组OD值均低于0.1μmol/LAngⅡ组,但仅0.1μmol/LAngⅡ+800μg/mLTMP组与之比较差异有统计学意义(P<0.05)。(2)0.1μmol/LAngⅡ组CFB上清液中Ⅰ型胶原含量高于空白对照组(P<0.01)。0.1μmol/LAngⅡ+800μg/mLTMP组CFB上清液中Ⅰ型胶原含量低于0.1μmol/LAngⅡ组(P<0.05)。(3)0.1μmol/LAngⅡ组Ⅰ型胶原mRNA表达高于空白对照组(P<0.05)。0.1μmol/LAngⅡ+800μg/mLTMP组Ⅰ型胶原mRNA表达低于0.1μmol/LAngⅡ组(P<0.05)。结论:川芎嗪可以抑制血管紧张素Ⅱ诱导的大鼠心肌成纤维细胞的增殖,减少Ⅰ型胶原的分泌与合成。
Objective： To observe the effects of tetramethylpyrazine （TMP） on the synthesis of rat cardiac fibroblasts （CFBs） induced by angiotensin Ⅱ mechanism of TMP in treating myocardial fibrosis. proliferation find type （Ang Ⅱ ）, and to Ⅰ collagen explore the Methods： CFBs were isolated from neonatal rats, and the fourth-passage CFBs were used in the entire test and were stimulated by 0.1μmol/L Ang 11 in vivo. The CFB proliferation was measured by methyl thiazolyl tetrazolium （MTT） assay. Type Ⅰ collagen in the cell culture supernatant was measured by enzyme-linked immunosorbent assay. The expression of mRNA of type Ⅰ collagen was semi-quantitatively measured by reverse transcription-polymerase chain reaction （RT-PCR）. Results： （1） In MTT assay, the optical density of CFBs cultured with 0.1μmol/L Ang Ⅱ was higher than that of the blank control cultured with 2% fetal bovine serum-Dulbecco＇s modified Eagle＇s medium （FBS-DMEM）. The difference was statistically significant （P 〈0.05）. Both optical densities of CFBs cultured with 0.1μmol/L Ang 11 plus 800 μg/mL TMP and 0.1μmol/L Ang Ⅱ plus 600 μg/mL TMP were lower than that of CFBs cultured with0.1μmol/L Ang Ⅱ , but only the difference between 0.1μmol/L Ang Ⅱ plus 800 μg/mL TMPgroupand0.1μmol/L Ang Ⅱ group was significant （P〈0.05）. （2） The content of type I collagen secreted by CFBs cultured with 0.1μmol/L Ang Ⅱ was higher than that with 2% FBS-DMEM （P〈0. 01）. The content of type I collagen secreted by CFBs cultured with 0.1μmol/L AngⅡ plus 800 μg/mL TMP was lower than that with 0.1μmol/LAng Ⅱ （P〈0.05）. （3） The level of type Ⅰ collagen mRNA in 0.1μmol/L Ang Ⅱ group was higher than that in blank control group, and lower than that in 0.1μmol/L Ang Ⅱplus 800 μg/mL TMP group. Both the differences between 0.1μmol/L AngⅡ group and the blank control group and between 0.1μmol/L Ang Ⅱ group and0.1μmol/L Ang Ⅱplus 800μg/mL TMP group were statistically significant （ P〈0. 05）. Conclusion： TMP can not only inhibit the proliferation of CFBs, but also decrease the secretion and the mRNA expression level of collagen Ⅰ in cultured CFBs of rat which are increased by Ang Ⅱ.
出处
《中西医结合学报》
CAS
2009年第3期232-236,共5页
Journal of Chinese Integrative Medicine
基金
国家自然科学基金资助项目(No.30200371)
