邢 燕,历 飞,张丽艳,刘 鸿,白 剑.欧前胡素通过ERK1/2信号保护心肌细胞低氧性损伤[J].,2018,(3):442-446 |
欧前胡素通过ERK1/2信号保护心肌细胞低氧性损伤 |
Imperatorin Protects H9c2 Cardiomyoblasts Cells from Hypoxia Induced Injury through Activation of ERK1/2 Signaling Pathway |
投稿时间:2017-10-10 修订日期:2017-10-30 |
DOI:10.13241/j.cnki.pmb.2018.03.009 |
中文关键词: 欧前胡素 心肌细胞 低氧 ERK1/2 凋亡 |
英文关键词: Imperatorin Cardiac myocytes Hypoxic Extracellular Signal-Regulated Kinase1/2 Apoptosis |
基金项目:辽宁省自然科学基金项目(201602506) |
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中文摘要: |
摘要 目的:研究欧前胡素对低氧诱导的大鼠心肌细胞损伤的保护作用和机制。方法:采用CO2-95%和N2-5%的细胞培养箱诱导H9c2大鼠心肌细胞建立心肌细胞低氧损伤模型,采用不同浓度的欧前胡素孵育细胞12、24 h,检测上清液中乳酸脱氢酶(lactate dehydrogenase,LDH)、超氧化物歧化酶(superoxide dismutase,SOD) 、丙二醛(malondialdehyde MDA)的含量。采用MTT方法检测细胞的存活率,Annexin V/PI双标记流式细胞术检测细胞凋亡比例,蛋白质印迹法检测检ERK1/2蛋白的表达。结果:低氧处理H9c2心肌细胞12 h后,上清中LDH和MDA的含量分别为(523.28±90.29) U/L和(5.59±0.33) U/L,均明显高于对照组(P<0.05),而SOD的含量[(12.23±1.38) U/mg]明显低于对照组(P<0.05)。低、高浓度欧前胡素孵育12 h后,细胞存活率分别为(64.51±2.78)%和(73.22±3.56)%,低、高浓度欧前胡素孵育24 h后,细胞存活率分别为(80.21±4.67)%和(87.38±5.41)%,均较与模型组显著升高(P<0.05)。高浓度欧前胡素孵育12 h和24 h后凋亡细胞比例分别为(39.67±4.11)%和(49.61±3.39)%,均较模型组显著降低(P<0.05),10 μmol/L的PD98059阻断ERK1/2信号通路后细胞存活率均较高浓度欧前胡素组显著降低,凋亡细胞比例较高浓度欧前胡素明显升高(P>0.05)。高浓度欧前胡素孵育12 h和24 h后,ERK1/2蛋白相对表达量分别为(1.92±0.09)和(2.42±0.21),与模型组相比均显著增加(P<0.05)。结论:欧前胡素可能通过活化ERK1/2信号通路保护低氧诱导的心肌细胞损伤。 |
英文摘要: |
ABSTRACT Objective: To study the protective effect and mechanism of imperatorin on the hypoxia induced injury of H9c2 car- diomyoblasts. Methods: H9c2 cardiomyoblasts were cultured in the incubator with CO2-95% and N2-5% to induce hypoxia injury model. The LDH, MDA and SOD were detected at 12 h and 24 h in the hypoxia incubator. MTT method was tested to measure the survival rate of cardiomyoblasts; the apoptosis ratio of H9c2 cardiomyoblasts was detected by Annexin v /PI double-labeled flow cytometry; the ex- pression of the ERK1/2 protein was detected by Western Blotting. Results: After culturing 12 h in the hypoxia incubator, the concentra- tions of LDH and MDA in supernatant were(523.28±90.29) U/L and (5.59±0.33) U/L respectively, and they were significantly higher than the control group (P<0.05); the concentration of SOD in supernatant was (12.23±1.38) U/mg respectively, and it was lower than the control group (P<0.05); after culturing 12 h in different concentrations of imperatorin, cell survival rates in low imperatorin and high im- peratorin group were (64.51±2.78)% and (73.22±3.56)% respectively, and after culturing 24 h in different concentrations of impera- torin, cell survival rates in low imperatorin and high imperatorin group were (80.21±4.67)% and (87.38±5.41)% respectively, and they were significantly higher than model group (P<0.05); after culturing 12 h and 24 h in high imperatorin group, apoptotic cells ratios were (39.67±4.11)% and (49.61±3.39)% respectively, and they were significantly lower than model group(P<0.05); after blocking ERK1/2 with 10 μmol/L PD98059, and cell survival rates were significantly lower than high imperatorin group, apoptotic cells ratios were signif- icantly higher than high imperatorin group (P>0.05); after culturing 12h and 24h in high imperatorin group, the expression of ERK1/2pro- tein were (1.92±0.09) and (2.42±0.21) respectively, and compared with model group they increased significantly(P<0.05). Conclusion: Imperatorin may protect H9c2 cardiomyoblasts cells from hypoxia induced injury through activation of ERK1/2 signaling pathway. |
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