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目的:研究云南鼠尾草提取物对大鼠H9c2心肌细胞缺氧/复氧损伤保护作用的机制。方法:建立大鼠H9c2心肌细胞缺氧/复氧损伤模型,并采用云南鼠尾草提取物进行干预。将体外培养的H9c2大鼠心肌细胞随机分为6组:正常对照(C)组、缺氧/复氧模型(H/R)组、缺氧/复氧模型+维拉帕米阳性对照(H/R+V)组、缺氧/复氧模型+云南鼠尾草低(H/R+L,0.01 mg·L~(-1))、中(H/R+M,0.1 mg·L~(-1))、高(H/R+H,1.0 mg·L~(-1))剂量组。采用噻唑蓝(MTT)法测定细胞存活率,利用检测试剂盒测定丙二醛(MDA)的含量和乳酸脱氢酶(LDH)的活性,通过荧光酶标仪测定荧光吸光度(A)值反映细胞内活性氧(ROS)水平。结果:与模型组比较,云南鼠尾草低、中、高剂量组均能显著提高心肌细胞存活率(P<0.05或P<0.01),云南鼠尾草高剂量组显著减少细胞内LDH漏出量(P<0.05或P<0.01)、胞浆内MDA的含量(P<0.01)和细胞内ROS水平(P<0.05)。结论:云南鼠尾草提取物对大鼠H9c2心肌细胞缺氧/复氧损伤具有保护作用,其相关作用机制可能与其减少脂质过氧化物和清除细胞氧自由基有关。
Objective: To investigate the protective effect of sage extract of Yunnan on rat H9c2 cardiomyocytes against hypoxia / reoxygenation injury. Methods: The hypoxia / reoxygenation injury model of rat H9c2 cardiomyocytes was established and the extracts of Yunnan sage were used for intervention. H9c2 rat cardiomyocytes cultured in vitro were randomly divided into 6 groups: normal control group (C), hypoxia / reoxygenation model (H / R) group, hypoxia / reoxygenation model + verapamil positive control (H / R + M, 0.1 mg · L ~ (-1)), hypoxia / reoxygenation model + (-1)), high (H / R + H, 1.0 mg · L -1) dose group. Cell viability was determined by MTT assay. Malondialdehyde (MDA) content and lactate dehydrogenase (LDH) activity were measured by a detection kit. Fluorescence absorbance (A) Internal reactive oxygen species (ROS) levels. Results: Compared with the model group, the survival rate of cardiomyocytes of Yunnan sage could be significantly increased (P <0.05 or P <0.01), and the high dose of Yunnan sage significantly reduced the amount of intracellular LDH leakage (P <0.05 or P <0.01), intracellular content of MDA (P <0.01) and intracellular ROS level (P <0.05). CONCLUSION: Scutellaria yunnanensis extract has a protective effect on H9c2 cardiomyocytes from hypoxia / reoxygenation injury. The possible mechanism may be related to the reduction of lipid peroxides and scavenging of oxygen free radicals.