阿齊沙坦對(duì)轉(zhuǎn)化生長(zhǎng)因子誘導(dǎo)的自發(fā)性高血壓大鼠血管平滑肌細(xì)胞增殖及遷移的影響

李慧等

[摘要]目的探討阿齊沙坦對(duì)轉(zhuǎn)化生長(zhǎng)因子_β（TGF_β）誘導(dǎo)的自發(fā)性高血壓大鼠血管平滑肌細(xì)胞（VSMCs）增殖及遷移的影響。方法采用組織塊貼壁培養(yǎng)法進(jìn)行血管平滑肌細(xì)胞的原代培養(yǎng)，取3～5代細(xì)胞用于實(shí)驗(yàn)。用MTT法測(cè)定細(xì)胞增殖情況，用Boyden小室測(cè)試細(xì)胞遷移情況。采用10ng/ml TGF_β刺激VSMCs，并用阿齊沙坦干預(yù)，24h后用Western blot和RT_PCR檢測(cè)VSMCs中MMP_2、MMP_9及TIMP_1蛋白和mRNA的表達(dá)。結(jié)果TGF_β可促進(jìn)大鼠VSMCs的增殖及遷移，并增加大鼠VSMCs的MMP_2、MMP_9和TIMP_1蛋白表達(dá)。阿齊沙坦則可抑制TGF_β誘導(dǎo)的大鼠VSMCs的增殖和遷移，并降低大鼠VSMCs的MMP_2、MMP_9及TIMP_1表達(dá)。結(jié)論阿齊沙坦可能通過下調(diào)MMP_2、MMP_9及TIMP_1的表達(dá)來抑制TGF_β誘導(dǎo)的自發(fā)性高血壓血管平滑肌細(xì)胞的增殖及遷移。

[關(guān)鍵詞]MMP_2；MMP_9；TIMP_1；血管平滑肌細(xì)胞；增殖；遷移；阿齊沙坦

中圖分類號(hào)：R544.1文獻(xiàn)標(biāo)識(shí)碼：A文章編號(hào)：1009_816X（2014）04_0274_04

[Abstract] Objective To investigate the effects of azilsartan on TGF_β_induced spontaneously hypertensive vascular smooth muscle cells （VSMCs） proliferation and migration. Methods Rat VSMCs were cultivated by the method of tissue explants adherence. Cells of generation 3 to 5 were used as the experimental system. The MTT test was used to measure cell proliferation and Boyden chamber assay was used to measure cell migration. Primary cultured VSMCs were treated by 10 ng/ml TGF_β and azilsartan for 24 hours. The expression of MMP_2， MMP_9， TIMP_1 were measured by Western blot and RT_PCR. Results The expression of MMP_2， MMP_9 and TIMP_1 in rat VSMCs stimulated by TGF_β increased significantly. VSMCs migration and proliferation also increased significantly. Azilsartan significantly inhibited TGF_β induced MMP_2， MMP_9 and TIMP_1 production in rat VSMCs， as well as VSMCs proliferation and migration. Conclusions Azilsartan inhibited TGF_β_induced VSMCs proliferation and migration by down_regulation the expression of MMP_2， MMP_9， TIMP_1.

[Key words] MMP_2； MMP_9； TIMP_1； Vascular smooth muscle cells ；Proliferation； Migration； Azilsartan

自發(fā)性高血壓是最常見的心血管疾病，以體循環(huán)動(dòng)脈壓升高為主要特點(diǎn)。高血壓伴發(fā)的血管重構(gòu)包括血管壁腔比增加、小動(dòng)脈稀少及血管功能異常。基質(zhì)金屬蛋白酶（matrix metalloproteinase，MMPs）是參與降解全身各種組織細(xì)胞外基質(zhì)（extracellular matrix，ECM）的蛋白酶家族，參與正常和病理?xiàng)l件下的組織重構(gòu)，金屬蛋白酶組織抑制物（TIMP）是MMPs的特異性抑制物[1]。MMPs/TIMP的動(dòng)態(tài)改變，造成選擇性的降解ECM，從而調(diào)整血管內(nèi)皮細(xì)胞形態(tài)、生長(zhǎng)和成活[2]。血管緊張素Ⅱ受體拮抗劑可能是通過抑制局部腎素血管緊張素醛固酮系統(tǒng)（RAS）活性和緩激肽的降解來抑制或逆轉(zhuǎn)血管重構(gòu)[3]。阿齊沙坦是一種血管緊張素Ⅱ受體拮抗劑，多用于治療高血壓病[4]。但沙坦類藥物是否可抑制血管平滑肌細(xì)胞（vascular smooth muscle cells，VSMCs）中MMPs/TIMP的表達(dá)則鮮有報(bào)道。……