目的: 探索石松的活性成分α-玉柏碱通过抗炎抗氧化作用对心肌缺血再灌注(MI/R)损伤的保护作用。方法: Wistar大鼠灌胃1 mL无菌水或含有2.5 mg·kg-1或5 mg·kg-1 α-玉柏碱的水,持续1周,雷米普利1 mg·kg-1作为阳性对照。结扎心脏左前降支30 min,再灌注3 h。超声检测射血分数EF(%)、短轴缩短分数FS(%)、左心室收缩压LVSP(mmHg)、左心室舒张末期压力LVEDP(mmHg)。伊文思蓝染指示心梗体积。试剂盒检测血清或心肌组织中总还原型谷胱甘肽(GSH)、乳酸脱氢酶(LDH)、天冬氨酸转氨酶(AST)、超氧化物歧化酶(SOD)和丙二醛(MDA)的水平。ELISA测定血清肌酸激酶-同工酶 (CK-Mb)、肌钙蛋白T (cTnT)、肿瘤坏死因子(TNF)-α、白介素(IL)-1β、IL-6和前列腺素E 2(PGE 2)。Western blotting检测环氧合酶2(COX-2)、金属基质蛋白酶(MMP)-2和MMP-9的蛋白表达。结果: 与假手术对照组比,MI/R组大鼠EF、FS和LVSP显著下降,而LVEDP明显升高(P<0.05)。α-玉柏碱能有效逆转这些超声心动指标(P<0.05)。α-玉柏碱还有效减少了MI/R大鼠的梗死面积和血清CK-Mb、cTnT、AST和LDH(P<0.05)。α-玉柏碱降低了MI/R升高的血清MDA、升高了降低的血清SOD和心肌GSH,减轻了氧化应激(P<0.05)。MI/R升高的血清炎症标志物TNF-α、IL-1β、IL-6、PEG2和心肌炎症标志物Cox-2均在5 mg·kg-1·d-1 α-玉柏碱处理下显著降低(P<0.05)。MI/R升高的MMP-2和MMP-9也被α-玉柏碱降低(P<0.05)。结论: α-玉柏碱能有效减轻氧化应激和炎症,改善MI/R导致的心肌损伤。对MMP-2和MMP-9的抑制可能是其作用机制。
Objective: To explore the protective effect of α-obscurine, an active component of Lycopodium on myocardial ischemia-reperfusion (MI/R) injury via anti-inflammatory and anti-oxidant ways. Methods: Wistar rats were intragastrically administered with 1 mL of sterile water or water containing 2.5 mg·kg-1 or 5 mg·kg-1 α-obscurine for 1 week. Ramipril 1 mg·kg-1 was used as a positive control. The left anterior descending branch of the heart was ligated for 30 min and reperfused for 3 h. Echocardiograph was employed to test the the ejection fraction (EF, %), short axis shortening fraction (FS, %), left ventricular systolic pressure (LVSP, mmHg) and left ventricular end diastolic pressure (LVEDP, mmHg). The volume of myocardial infarction was indicated with Evans blue staining. The kits detect total reduced glutathione (GSH), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), superoxide dismutase (SOD) and malondialdehyde (MDA) in serum or myocardial tissue. ELISA was used to test serum creatine kinase-isoenzyme (CK-Mb), troponin T (cTnT), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6 and prostaglandin E2 (PGE2). Western blotting detects the protein expression of cyclooxygenase 2 (COX-2), metal matrix protease (MMP)-2 and MMP-9. Results: Compared with sham operation control group, EF, FS, and LVSP in MI/R group were significantly decreased, while the LVEDP was significantly increased (P<0.05). α-obscurine could effectively reverse these echocardiographic indexes (P<0.05). α-obscurine also effectively reduced the infarct size of MI/R rats and the serum indicators of myocardial injury such as CK-Mb, cTnT, AST and LDH (P<0.05). α-obscurine reduced the serum MDA with elevated MI/R, increased the reduced serum SOD and myocardial GSH, and alleviated the oxidative stress in MI/R rats (P<0.05). Elevated serum inflammation markers TNF-α, IL-1β, IL-6, PEG2 and myocardial inflammation marker Cox-2 were significantly reduced under 5 mg·kg-1·d-1 α-obscurine treatment (P<0.05). It was further found that metalloproteinase (MMP)-2 and MMP-9 with elevated MI/R were also reduced by α-obscurine (P<0.05). Conclusion: Our results show that α-obscurine could effectively reduce oxidative stress and inflammation, and improve myocardial damage caused by MI/R. The inhibition of MMP-2 and MMP-9 may be its mechanism of action.
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