酒制豨莶草和生豨莶草对缺氧损伤的H9c2心肌细胞保护作用的比较研究  

Protective Effects of Wine-Processed and Raw Siegesbeckiae Herba on H9c2 Myocardial Cells Induced by Hypoxia:A Comparative Analysis

作　　者：白云绮 李慧[2] 高照 范冬冬 娄利霞[1] 吴爱明[1] 孙克寒 商洪才[1] 聂波[1,2] BAI Yunqi;LI Hui;GAO Zhao;FAN Dongdong;LOU Lixia;WU Aiming;SUN Kehan;SHANG Hongcai;NIE Bo(Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing,Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China;School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学东直门医院中医内科学教育部和北京市重点实验室,北京100700 [2]北京中医药大学中药学院,北京100029

出　　处：《世界中医药》2022年第6期779-783,789,共6页World Chinese Medicine

基　　金：第二批国家“万人计划”科技创新领军人才特殊支持经费资助项目(W02020052);北京中医药大学重点实验室项目(1000061222482)。

摘　　要：目的:比较酒制豨莶草和生豨莶草对大鼠心肌细胞缺氧损伤的保护作用,并探讨其作用机制,为临床用药选择提供实验依据。方法:采用H9c2细胞缺氧模型,给予不同浓度酒制豨莶草和生豨莶草干预,CCK-8法测定细胞成活率,比色法检测细胞培养上清液中乳酸脱氢酶(LDH)活性,超氧化物歧化酶(SOD)活性;荧光酶标仪检测细胞内活性氧(ROS)含量;用蛋白质印迹法(Western Blotting)检测凋亡B细胞淋巴瘤-2(Bcl-2)蛋白和Bcl-2相关X蛋白(Bax)的表达。结果:缺氧16 h成功制备H9c2心肌细胞缺氧损伤模型;筛选确定300μg/mL是生豨莶草和酒制豨莶草最佳给药浓度;与模型组比较,酒制豨莶草和生豨莶草均可提高细胞成活率(P<0.05),改善损伤细胞形态,减轻LDH外漏(P<0.05),降低ROS的含量(P<0.05),增加SOD活性(P<0.05),上调Bcl2(P<0.05)和Bcl-2/Bax的表达。给药浓度为300μg/mL时,生豨莶草在提高SOD活力和细胞成活率方面优于酒制豨莶草(P<0.05),酒制豨莶草在抑制LDH外漏的作用强于生豨莶草,在活性氧(ROS)含量和Bcl、Bax蛋白表达方面,差异无统计学意义(P>0.05)。结论:酒制豨莶草和生豨莶草对缺氧损伤的H9c2心肌细胞具有保护作用,其机制可能是通过抵抗氧化应激,抑制心肌细胞凋亡。Objective:To compare the protective effects of wine-processed and raw Siegesbeckiae Herba on H9c2 myocardial cells induced by hypoxia and explore the underlying mechanism to provide the experimental basis for clinical drug selection.Methods:Hypoxia-induced H9c2 cells were treated by wine-processed and raw Siegesbeckiae Herba of different concentrations.Cell viability was detected by Cell Counting Kit-8(CCK-8)and the lactic dehydrogenase(LDH)activity in cell culture supernatant by colorimetry.Superoxide dismutase(SOD)activity was also detected.The fluorescence microplate reader was used to detect the content of intracellular reactive oxygen species(ROS).Western blot was used to detect the protein expression of B-cell lymphoma 2(Bcl-2)and Bcl-2-associated X protein(Bax).Results:The hypoxia model was properly induced in H9c2 myocardial cells after 16 hours of hypoxia.The screening results revealed that the optimal concentration of wine-processed and raw Siegesbeckiae Herba was 300μg/mL.Compared with the conditions in the model group,wine-processed and raw Siegesbeckiae Herba could improve cell viability(P<0.05),improve damaged cell morphology,reduce LDH leakage(P<0.05),decrease ROS content(P<0.05),potentiate SOD activity(P<0.05),and up-regulate the expression of Bcl-2(P<0.05)and Bcl-2/Bax.In the context of administration concentration of 300μg/mL,raw Siegesbeckiae Herba was superior to wine-processed Siegesbeckiae Herba in potentiating SOD activity and increasing cell viability(P<0.05),while wine-processed Siegesbeckiae Herba was superior to raw Siegesbeckiae Herba in inhibiting LDH leakage.No statistically significant difference in ROS content and protein expression of Bcl and Bax was observed between wine-processed and raw Siegesbeckiae Herba.Conclusion:Wine-processed and raw Siegesbeckiae Herba possessed protective effects on H9c2 myocardial cells induced by hypoxia,and the mechanism might be related to the resistance to oxidative stress and the inhibition of myocardial apoptosis.

关 键 词：酒制豨莶草 生豨莶草 H9c2心肌细胞缺氧模型 心肌保护 氧化应激 细胞凋亡 乳酸脱氢酶 活性氧 

分 类 号：R285.5[医药卫生—中药学]