本文選題：PKC + 阿片藥物��； 參考：《中國(guó)醫(yī)院藥學(xué)雜志》2017年16期

【摘要】：目的:探討蛋白激酶C(PKC)信號(hào)通路及心肌縫隙連接蛋白Cx43改變?cè)诎⑵幬镱A(yù)處理中的作用。方法:原代心肌細(xì)胞分離培養(yǎng)。將培養(yǎng)5 d的心肌細(xì)胞分成8組。正常對(duì)照組(C組)不給予任何處理,建立培養(yǎng)乳鼠心肌細(xì)胞缺氧/復(fù)氧損傷模型(I/R組),嗎啡組(MF組)加入嗎啡至終濃度0.3μmol·L~(-1),舒芬太尼組(SF組)加入舒芬太尼至終濃度0.000 3μmol·L~(-1),PKC激動(dòng)劑PMA+嗎啡組(PMA+MF組)加入PMA至終濃度0.02μmol·L~(-1),再進(jìn)行嗎啡預(yù)處理;PKC抑制劑Rottlerin+嗎啡組(ROT+MF組)加入Rottlerin至終濃度5μmol·L~(-1),再進(jìn)行嗎啡預(yù)處理;PKC激動(dòng)劑PMA+舒芬太尼組(PMA+SF組)加入PMA至終濃度0.02μmol·L~(-1),再進(jìn)行舒芬太尼預(yù)處理;PKC抑制劑Rottlerin+舒芬太尼組(ROT+SF組)加入Rottlerin至終濃度5μmol·L~(-1),再進(jìn)行舒芬太尼預(yù)處理。各組做上述相應(yīng)處理后取材檢測(cè)細(xì)胞存活率。免疫熒光共聚焦技術(shù)檢測(cè)縫隙連接蛋白Connexin 43(Cx43)的平均光密度(AOD),用Western-blot檢測(cè)細(xì)胞Cx43總蛋白及其磷酸化水平(P-Cx43)的表達(dá)量。結(jié)果:與C組比較,其余各組心肌細(xì)胞存活率、Cx43 AOD值、心肌細(xì)胞Cx43總蛋白表達(dá)及P-Cx43表達(dá)降低(P0.05);與I/R組比較,MF組、SF組、ROT+MF組、ROT+SF組、PMA+MF組、PMA+SF組心肌細(xì)胞存活率、Cx43 AOD值、Cx43總蛋白和P-Cx43表達(dá)增高(P0.05);與MF組比較,ROT+MF組心肌細(xì)胞存活率、Cx43 AOD值、Cx43總蛋白及P-Cx43表達(dá)降低(P0.05),SF組心肌細(xì)胞存活率、Cx43 AOD值和Cx43總蛋白降低(P0.05),而P-Cx43表達(dá)增高(P0.05),PMA+MF組心肌細(xì)胞存活率、Cx43 AOD值、Cx43總蛋白和P-Cx43表達(dá)均增高(P0.05);ROT+SF組較SF組心肌細(xì)胞Cx43 AOD值、Cx43總蛋白和P-Cx43表達(dá)低(P0.05),而PMA+SF組較SF組心肌細(xì)胞存活率、心肌細(xì)胞Cx43 AOD值、Cx43總蛋白和P-Cx43表達(dá)高(P0.05)。結(jié)論:嗎啡和舒芬太尼預(yù)處理可減輕心肌細(xì)胞缺氧/復(fù)氧損傷,且嗎啡對(duì)心肌細(xì)胞的保護(hù)作用較舒芬太尼強(qiáng),這種心肌細(xì)胞保護(hù)作用可能與PKC信號(hào)通路的激活有關(guān)。
[Abstract]:Aim: to investigate the role of protein kinase C kinase (PKC) signaling pathway and cardiac gap junction protein (Cx43) in opioid preconditioning. Methods: primary cardiomyocytes were isolated and cultured. Cardiomyocytes cultured for 5 days were divided into 8 groups. Normal control group (group C) is not given any treatment, The model of hypoxic / reoxygenation injury of cultured neonatal rat cardiomyocytes was established. I / R group, morphine group (MF group)) were added morphine to the final concentration of 0. 3 渭 mol / L, and sufentanil group (SF group) were added sufentanil to the final concentration of 0.000 渭 mol / L PMAMF). The final concentration of PMA was 0. 02 渭 mol / L ~ (-1), and then the morphine preconditioning was performed with Rottlerin, the inhibitor of PKC. The rot MF group was added with Rottlerin to the final concentration of 5 渭 mol L ~ (-1). Then the morphine pretreatment with PMA sufentanil agonist PMA sufentanil was used to add PMA to the final concentration of 0. 02 渭 mol L ~ (-1), and then to do sufentanil (0. 02 渭 mol L- 1). Rottlerin sufentanil group was pretreated with sufentanil (5 渭 mol L ~ (-1)) and then sufentanil was pretreated with sufentanil (5 渭 mol 路L ~ (-1)). The cell survival rate was measured after the above treatment. The average optical density of gap junction protein (Connexin 43 Cx43) was detected by immunofluorescence confocal technique, and the expression of Cx43 total protein and its phosphorylation level (P-Cx43) was detected by Western-blot. Results: compared with group C, the survival rate of cardiac myocytes in other groups was determined by Cx43 AOD. Compared with the I / R group, the total protein expression of Cx43 and the expression of P-Cx43 in cardiomyocytes were decreased (P0.05); compared with the I / R group, the expression of Cx43 AOD and the expression of Cx43 total protein and P-Cx43 were increased in the myocytes of the MF group compared with that of the group of MF, and the myocardial fine cells in the group of row MF were compared with those in the group of MF, and the expression of Cx43 total protein and P-Cx43 was increased in the group of PMA-SF than in the group of MF. The expression of Cx43 total protein and P-Cx43 decreased in P0.05 / SF group, while the expression of Cx43 AOD and P-Cx43 in P0.05 / PMA-SF group was significantly higher than that in P0.05 / SF group. The expression of Cx43 total protein and Cx43 protein in SF group were significantly higher than that in P0.05 group (P < 0.05), while the expression of Cx43 AOD and Cx43 total protein and P-Cx43 in SF group were significantly higher than those in P0.05 group (P < 0.05). The expression of Cx43 total protein and P-Cx43 in SF group was significantly higher than that in P0.05 group. Compared with SF group, the expression of Cx43 total protein and P-Cx43 in SF group was lower than that in SF group, but the survival rate of PMA SF group was higher than that of SF group. The expression of Cx43 total protein and P-Cx43 in cardiomyocytes was high (P 0.05). Conclusion: morphine and sufentanil preconditioning can attenuate hypoxia / reoxygenation injury of cardiomyocytes and the protective effect of morphine on cardiomyocytes is stronger than sufentanil which may be related to the activation of PKC signaling pathway.
【作者單位】： 北京市海淀婦幼保健院麻醉科;寧夏醫(yī)科大學(xué)總醫(yī)院麻醉科;寧夏醫(yī)科大學(xué);
【基金】：國(guó)家自然科學(xué)基金(編號(hào):81260029)
【分類號(hào)】：R614

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