【摘要】：目的:探索高糖對(duì)大鼠膀胱平滑肌細(xì)胞(bladder smooth muscle cells,BSMCs)中鈣庫操控的鈣離子通道(store-operated calcium channel,SOCC)的影響。通過高糖處理BSMCs,模擬糖尿病膀胱(diabetes cystopathy,DCP)體外環(huán)境,探究BSMCs中SOCC的相關(guān)變化,為DCP的病因?qū)W及其治療提供新的思路。方法:使用Ⅱ型膠原酶分離S-D大鼠BSMCs,并用α-SMA進(jìn)行免疫熒光鑒定。分離的原代BSMCs進(jìn)行培養(yǎng)傳代。將傳至第3-5代的BSMCs同步化后根據(jù)培養(yǎng)基成分及濃度分為:正常葡萄糖濃度組(葡萄糖5.5mmol/L,NG組),甘露醇高滲組(葡萄糖5.5mmol/L+甘露醇19.5mmol/L,OC組),高葡萄糖濃度組(葡萄糖25mmol/L,HG組)三組,于37℃、5%CO2的孵箱中培養(yǎng)2.5天。各組BSMCs負(fù)載Fluo-4AM,通過激光共聚焦顯微鏡觀察記錄三組BSMCs激活(CPA,10umol/L)和抑制SOCC(SKF-96365,10umol/L)后細(xì)胞內(nèi)熒光強(qiáng)度值,計(jì)算其熒光強(qiáng)度值(F1)與背景熒光強(qiáng)度值(F0)的比值(F1/F0)來比較各組間鈣離子濃度的變化。通過RT-qPCR法和Western blot法檢測(cè)三組BSMCs中SOCC相關(guān)蛋白STIM1和Orai1的表達(dá)情況。結(jié)果:經(jīng)過α-SMA免疫熒光鑒定證實(shí)成功分離培養(yǎng)出膀胱平滑肌細(xì)胞。HG組加入SOCC激動(dòng)劑CPA后較NG組、OC組鈣離子濃度的顯著增加(2.105±0.105,P0.05),加入SOCC抑制劑SKF-96365后較NG組、OC組鈣離子濃度的顯著降低(1.610±0.109,P0.05),NG組與OC組之間無明顯差異(P0.05);HG組BSMCs中STIM1和Orai1的mRNA表達(dá)水平以及蛋白表達(dá)水平明顯高于NG組、OC組(P0.05),NG組與OC組之間無明顯差異(P0.05)。結(jié)論:經(jīng)過2.5天高糖作用下能激活大鼠膀胱平滑肌細(xì)胞的SOCC,可能參與組成細(xì)胞外鈣離子內(nèi)流部分,從而參與相應(yīng)的病理生理機(jī)制。另外,在高糖作用下,BSMCs中SOCC組成分子STIM1和Orai1表達(dá)水平發(fā)生改變可能為DCP的病理機(jī)制提供體外實(shí)驗(yàn)依據(jù),從而為DCP病因?qū)W研究提供新的思路。但高糖對(duì)SOCC的作用機(jī)制尚不清楚,仍需后續(xù)研究。
[Abstract]:Objective: To explore the effect of high glucose on calcium channel (SOCC) in the control of the calcium pool in the bladder smooth muscle cells (BSMCs) of the rat. The changes of SOCC in BSMCs were investigated by high-glucose treatment of BSMCs, and the changes of SOCC in BSMCs were investigated. Methods: The type 鈪，

本文編號(hào)：2457103