爲研究無乳鏈球菌（Streptococcus agalactiae，S呢在. agalactiae）作用奶牛乳腺成(chéng)纖維細胞（BMFBs）爸能後(hòu)對(duì)MMPs/TIMPs與uPA系統（uP愛很A/PAI-1）表達的影響，以進(jìn)一步闡明MMPs/TIMPs與u月問PA系統在調控細胞外基質（ECM）代謝的作市朋用，將(jiāng)107 cfu/m音嗎L熱滅活S. agalactiae菌液作不車用于BMFBs，在作用後(hòu)不同時(shí)間點（6、1這離2、24、48 h）通過(guò)RT-qPCR和West報中ern Blot檢測MMP-2、MMP-9、TIMP-1、TI議要MP-2、uPA、PAI-1 mRNA轉錄水平和相應蛋白表達水平，并秒山用明膠酶譜法檢測MMP-2、MMP-9的酶活性。結果顯示：熱雪外滅活S. agalactiae作用後(hòu)，MMP-9、TIM制但P-2、uPA、PAI-1 mRNA轉錄水平均有不同吃影程度下降趨勢，但MMP-2 mRNA水平上調，TIMP-1 mRNA水平呈先村懂上升後(hòu)下降趨勢；明膠酶譜也懂法檢測MMP-2的酶活性較MMP-9明顯，二者均在作用6 h、48 h活性最強理秒；MMP-2和MMP-9蛋白表達內線水平與對(duì)照組相比主要呈先升高後(hòu)下降趨勢煙區，但在作用後(hòu)期MMP-2蛋白表達又出現了回升生自趨勢；TIMP-1蛋白水平與對(duì)照組相比主子弟要呈先下降後(hòu)上升趨勢，在作用前期與MMP-9蛋白用裡表達相反，TIMP-2蛋白表達與對(duì)照組相比呈上升趨勢；uPA和科笑PAI-1蛋白水平與對(duì)照組相拍月比均呈先降低後(hòu)升高再降低趨勢，均在24 h達到峰值黑資，随後(hòu)呈下降趨勢。研究表明，熱滅活S. agalactiae喝業菌液可誘導BMFBs中MMPs/TIMPs表達，影響uPA系統參與呢多調控MMPs的表達及活性。
The Effect of Inactivated S. ag亮就alactiae on Metalloproteinase an志新d uPA System Exp作動ression in BMFBs
In order to study the effect of道中 Streptococcus agalactiae（S. agalacti我事ae）on bovine mammary fibroblasts（BM通習FBs）and subsequently on the 匠歌expression of MMP錢票s/TIMPs and uPA system（uPA/PAI-1）謝還，and to further clarify t通購he role of MMPs/TIMPs and uPA system 新為in regulating extracell算唱ular matrix（ECM）metaboli慢技sm，107cfu/mL heat-inactivated謝水 S. agalactiae was a熱喝cted on BMFBs，the levels of MMP-用冷2，MMP-9，TIMP-1，TIMP-2，uP樹票A，PAI-1 mRNA transcri她們ption and corresponding protein expre票相ssion were detected b討暗y RT-qPCR and Western 藍理Blot at different time poi公畫nts（6，12，24 and 48 h），then the enz離吃ymatic activities of MMP-日好2 and MMP-9 were detected by草呢 gelatin zymography. The r刀自esults showed that after the rea志站ction of heat-inact子唱ivated S. agalactiae，th舊的e level of transcription of MMP-9朋電，TIMP-2，uPA and PAI-1 mRNA trende銀物d to reduce to different extent，聽了except that of MMP-2 mRNA，and tha海舊t of TIMP-1 mRN從嗎A increased firstly and then de白船creased；the enzymatic activity o畫是f MMP-2 was mor錢媽e obvious than that離為 of MMP-9 as detected by gelatin zymo用離graphy，both of which were stro一業ngest in 6 and 8 h after reaction；a中議s compared with 匠聽the control group，the level東家 of protein expression of MMP-2 and M黃也MP-9 trended to increase件影 prior to decrease，but the p美制rotein expressi就窗on of MMP-2 rose again at la話子ter stage；as compared t林綠o the control group，the level of protei光學n of TIMP-1 sho一科wed a trend of decreasing p相道rior to rising，which was 離睡contrary to that of MMP-9 at ea信動rly stage，and TIMP-2 trended to也很 increase；the levels of 費錯protein of uPA and少了 PAI-1 both decreased first 快木then increased，and d討水ecreased finally，喝有reaching the peak at 24 h after r路雨eaction. In conclusion，the 地麗expression of MMPs/TIMPs in 的農BMFBs could be induc醫場ed by heat-inactivated S. agalac生農tiae，which could店亮 influence uPA system 窗喝in regulating the expression a房少nd activity of MMPs.
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