目的 評估丹參素對心肌缺血/再灌注（MI/R）損傷的保護(hù)作用及其可能機(jī)制。方法 32只大鼠隨機(jī)分為假手術(shù)組、模型組組、丹參素30 mg/kg組和丹參素60 mg/kg組，丹參素組在再灌注開始后5 min內(nèi)iv相應(yīng)劑量丹參素，假手術(shù)組和模型組注射等量生理鹽水；再灌注3 h后，向冠狀動脈中注入2 mL的3%伊文思藍(lán)染料；取大鼠動脈血和心臟，通過組織化學(xué)測定心肌梗死面積大小，通過ELISA測定血清中的肌酸激酶-同工酶（CK-MB）和心肌肌鈣蛋白（cTnI）水平。通過氧糖剝奪/恢復(fù)（OGD/R）H9c2心肌細(xì)胞模型評估丹參素預(yù)處理對細(xì)胞的直接作用，使用MTT、ELISA和流式細(xì)胞術(shù)分別測定丹參素對細(xì)胞活力、乳酸脫氫酶（LDH）釋放和細(xì)胞凋亡的影響，使用RT-PCR和Western blotting免疫印跡測定miR-199a-5p及其下游蛋白Akt和ERK1/2的表達(dá)。結(jié)果 丹參素治療可顯著降低心肌梗死面積以及血清中CK-MB和cTnI的水平（P<0.05）；丹參素預(yù)處理顯著改善OGD/R心肌細(xì)胞活力恢復(fù)能力，減少細(xì)胞凋亡水平，顯著提高Akt和ERK1/2的mRNA和蛋白表達(dá)水平，降低miR-199a-5p的mRNA表達(dá)水平（P<0.05）；上調(diào)miR-199a-5p顯著降低Akt和ERK1/2的mRNA和蛋白表達(dá)水平（P<0.05），而下調(diào)miR-199a-5p可顯著提高Akt和ERK1/2的mRNA和蛋白表達(dá)水平（P<0.05）。結(jié)論 丹參素可以通過抑制miR-199a-5p增加Akt和ERK1/2表達(dá)來發(fā)揮心臟保護(hù)作用。

Objective To evaluate the protective effects of Danshensu, the effective component of Salvia miltiorrhiza (Danshen), against myocardial ischemia/reperfusion (MI/R) injury in cardiomyocytes and its possible underlying mechanism. Methods 32 rats were randomly divided into four groups: sham operation group, model group, Danshensu 30 mg/kg group and Danshensu 60 mg/kg group. Danshensu 30 mg/kg group and Danshensu 60 mg/kg group were given all drugs intravenously within 5 min after reperfusion, sham operation group and model group were injected with the same amount of physiological saline. Three hours after reperfusion, 2 mL of 3% Evans was injected into the coronary artery. The size of myocardial infarction was determined by histochemistry. The levels of creatine kinase MB (CK-MB) and cardiac troponin (cTnI) in serum were measured by ELISA. In addition, the direct toxicity of Danshensu on cardiomyocytes was evaluated by oxygen glucose deprivation/recovery (OGD/R) model. MTT, ELISA and flow cytometry were used to determine the effects of Danshensu on cell viability, lactate dehydrogenase (LDH) release and apoptosis. RT-PCR and Western blotting were used to detect the effects of Danshensu on cell viability, LDH release and apoptosis. The expression of miR-199a-5p and its downstream proteins Akt and ERK1/2 were detected by Western blotting. Results Danshensu treatment significantly reduced myocardial infarction area and serum CK-MB and cTnI levels (P<0.05); Danshensu pretreatment significantly improved the viability of OGD/R myocardial cells, reduced the level of apoptosis, significantly increased the mRNA and protein expression levels of Akt and ERK1/2, and decreased the mRNA expression level of miR-199a-5p(P<0.05). Up regulation of miR-199a-5p significantly decreased the mRNA and protein expression levels of Akt and ERK1/2 (P<0.05), and down-regulation of miR-199a-5p significantly increased the mRNA and protein expression levels of Akt and ERK1/2 (P<0.05).Conclusion Danshensu provides the cardioprotective effects against MI/R injury via increasing Akt and ERK1/2 through inhibition of miR-199a-5p.

R285.5

國家重點研發(fā)項目（2017YFC1307602）；天津市科技計劃項目（15ZXLCSY00040）