南葶苈子水提物对多柔比星诱导H9c2细胞凋亡和氧化应激的抑制作用

冯卫生, 杨方方, 张莉, 杨雁芸, 白志尧, 李孟, 樊慧, 栗萌萌, 郑晓珂

中国药学杂志 ›› 2018, Vol. 53 ›› Issue (23) : 1999-2007.

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中国药学杂志 ›› 2018, Vol. 53 ›› Issue (23) : 1999-2007. DOI: 10.11669/cpj.2018.23.005
论著

南葶苈子水提物对多柔比星诱导H9c2细胞凋亡和氧化应激的抑制作用

  • 冯卫生1,2, 杨方方1, 张莉1,2, 杨雁芸1, 白志尧1, 李孟1,2, 樊慧1, 栗萌萌1, 郑晓珂1,2*
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Protective Effects of Aqueous Extract from Descurainia sophia on Doxorubicin-Induced Cardiomyocyte Injuryvia Suppressing Apoptosis and Oxidative Stress

  • FENG Wei-sheng1,2, YANG Fang-fang1, ZHANG Li1,2, YANG Yan-yun1, BAI Zhi-yao1, LI Meng1,2, FAN Hui1, LI Meng-meng1, ZHENG Xiao-ke1,2*
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摘要

目的 探明南葶苈子水提物对多柔比星(doxorubicine,Dox)引起的H9c2心肌细胞损伤的保护作用及潜在机制。方法 应用5 μg·mL-1 Dox处理H9c2细胞诱导建立心肌细胞损伤模型,分为对照组、模型组、阳性组、南葶苈子水提物不同剂量给药组,检测细胞活力,凋亡率,线粒体膜电位,细胞活性氧(ROS)、总超氧化物歧化酶(T-SOD)、乳酸脱氢酶(LDH)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-PX)水平,凋亡通路关键蛋白p53、Bax、Bcl-2、Caspase-3蛋白表达水平,使用UPLC-MS联用的方法分析鉴定南葶苈子水提物中主要成分峰。结果 南葶苈子水提物能提高H9c2细胞活力(P<0.01),降低凋亡水平(P<0.01),改善线粒体膜电位水平与ROS水平(P<0.05),改善细胞内SOD及GSH-PX酶活力、LDH、MDA水平(P<0.01或P<0.05),调节凋亡通路关键蛋白caspase-3、Bax/Bcl-2、p53蛋白表达水平(P<0.01或P<0.05),同时,共分析鉴定了南葶苈子水提物中7种含量最高的成分。结论 南葶苈子水提物可以有效保护由Dox引起的H9c2细胞损伤,其机制可能与改善细胞的氧化应激,抑制内源性凋亡通路,而抑制细胞凋亡的发生有关。

Abstract

OBJECTIVE To study the protective effects and mechanisms of aqueous extract from Descurainia sophia on doxorubicin(Dox)-induced cardiomyocyte injury.METHODS The Dox induced H9c2 cell apoptotic model was established, then the cells were divided into normal group (NC), model group (Dox), positive group (resveratrol,RSV), and different doses of aqueous extract of Descurainia sophia (DS). The viability of H9c2 cells was detected by MTT assay. The apoptosis rate, mitochondrial membrane potential and reactive oxygen species (ROS) level were detected by flow cytometry. The levels of T-SOD, LDH, MDA and GSH-PX were measured. And the protein expression levels of caspase-3, Bcl-2, Bax and p53 were detected by western blot, HPLC-MS identification of aqueous extract from Descurainia sophia.RESULTS After treating with DS products, the survival rate of H9c2 was increased (P<0.01), the apoptosis rate was significantly decreased (P<0.01),mitochondrial membrane potential was significantly increased (P<0.01), the level of ROS was significantly decreased (P<0.05), T-SOD and GSH-PX activities were significantly increased (P<0.01), the levels of LDH and MDA content were significantly decreased(P<0.01). Moreover, DS reduced the expression of caspase-3 (P<0.01), regulated the expression of Bax/Bcl-2 (P<0.01), decreased the expression of p53 (P<0.05). Seven components were identified from DS by HPLC/MS analysis.CONCLUSION DS can effectively protect cardiomyocyte, and its mechanism is probably associated with correcting functional disorders of oxidative stress of cardiomyocytes, and inhibit mitochondrial apoptotic pathway.

关键词

南葶苈子 / H9c2细胞 / 凋亡 / 氧化应激

Key words

Descurainia sophia / H9c2 cells / apoptosis / oxidative stress

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冯卫生, 杨方方, 张莉, 杨雁芸, 白志尧, 李孟, 樊慧, 栗萌萌, 郑晓珂. 南葶苈子水提物对多柔比星诱导H9c2细胞凋亡和氧化应激的抑制作用[J]. 中国药学杂志, 2018, 53(23): 1999-2007 https://doi.org/10.11669/cpj.2018.23.005
FENG Wei-sheng, YANG Fang-fang, ZHANG Li, YANG Yan-yun, BAI Zhi-yao, LI Meng, FAN Hui, LI Meng-meng, ZHENG Xiao-ke. Protective Effects of Aqueous Extract from Descurainia sophia on Doxorubicin-Induced Cardiomyocyte Injuryvia Suppressing Apoptosis and Oxidative Stress[J]. Chinese Pharmaceutical Journal, 2018, 53(23): 1999-2007 https://doi.org/10.11669/cpj.2018.23.005
中图分类号: R965   

参考文献

[1] GU G G, YANG P J. Shennong′s Classic of Materia Medica(神农本草经). Beijing:Xueyuan Publishing House, 2007:240.
[2] LI S Z. Compendium of Materia Medica(本草纲目). Beijing:People′s Medical Publishing House,1987:530-935.
[3] Ch.P(2015)Vol Ⅰ(中国药典2015年版.一部). 2015:333.
[4] LI Y Y, ZHANG F B, LUO L H, et al. Research on Semen Lepidii seu Descurainiae Powder in treatment of chronic heart failure. Chin Foreign Med Res(中外医学研究),2010,8(5):53.
[5] PAN J Y, JIN Z G, WU F H. Research advances in Semen Lepidii seu Descurainiae and its formulae for cardiovascular diseases. Shanghai J Tradit Chin Med(上海中医药杂志),2008,42(12):83-85.
[6] ZHANG W,ZHANG Y,LI H. Application of Semen Lepidii seu Descurainiae Powder in the treatment of congestive heart failure. World J Integr Tradit West Med(世界中西医结合杂志), 2010,5(4):349.
[7] GUO J,CHEN C X. Influnce of Tinglizi on some neuroendocrine factors and type Ⅰ and Ⅲ collagen in ventricular remodeling induced by abdominal aortic banding in rats. J Chin Med Mater(中药材), 2007,30(8):963-967.
[8] GUO J, CHEN C X, DU J, et al. Semen descurainiae inhibits CYP11B1,CYP11B2 and TGF-β-1 mRNA expression in left ventricular. J Chin Med Mater(中药材),2008,31(11):1691-1695.
[9] GUO J, CHEN C X, SHEN Y H, et al. Effect of aqueous extract from Semen Lepidii seu Descurainiae on ventricular remodeling in experimental animals. Chin Tradit Herb Drugs(中草药),2007,38(10):1519-1523.
[10] GUO J, CHEN C X, GU W L, et al. Influence of Tinglizi on collagen volume fraction and perivascular collagen area in left ventricle tissue of cardiac hypertrophy induced by abdominal aortic banding in rats. China J Chin Mater Med(中国中药杂志), 2008,33(3):284-287.
[11] FANG Z J, XIONG X D. Effects of helveticoside on hemodynamics expression in rats with MTC-induced pulmonary hypertension. Pract Clin J Integr Tradit Chin West Med(实用中西医结合临床),2004,4(5):73-74.
[12] WANG Q C, LIU G F, LI C C. Cardiac effect, bioactivity, desorption, accumulation and toxicity of Descurainia semen. J Fujian Med Univ(武汉医学院学报), 1964(Z1):27-33.
[13] WU X L, YANG Y Z, HUANG D L. Effect of aqueous extract of Lepidium apetalumon dog′s left ventricular function. J Chin Med Mater(中药材), 1998,21(5):243-245.
[14] ANGSUTARARUX P, LUANPITPONG S, ISSARAGRISIL S. Chemotherapy-induced cardiotoxicity: overview of the roles of oxidative stress. Oxid Med Cell Longev,2015,2015:795602.
[15] SUN K. Studies of constituents and biological activities of Descurainiae sophia. Shenyang:Shenyang Pharmaceutical University,2005.
[16] ZHOU X, TANG L, WU H, et al. Chemometric analyses for the characterization of raw and processed seeds of Descurainia sophia (L.) based on HPLC fingerprints. J Pharm Biomed Anal,2015,111:1-6.
[17] MENG Z H, QIAO L, WHANG Z Q, et al.Analysis of the constituents in Semen Descurainiae by UPLC/Q-TOF-MS/MS. J Chin Pharm Sci, 2015, 24(5):303-309.
[18] WANG A Q, WHANG X K, ZHAO H Y, et al. Studies on chemical composition and the quality of Descurainiae sophia. Chin Remed Clin (中国药物与临床), 2005,5(1):5-6.
[19] FENG Z Y, WANG X L, ZHNAG X K. Herbal textual research on Semen Lepidii seu Descurainiae. Mod Tradit Chin Med Mater Med-World Sci Technol(世界科学技术-中医药现代化), 2014,16(16):1938-1941.
[20] ZHANG G S, BAI Y P, WANG X L, et al.Effective fractions and mechanism of descurainiae lepidii semen in chronic heart failure rats. China J Exp Tradit Med Form(中国实验方剂学杂志), 2017, 23(4):118-125.
[21] ABE J, YAMADA Y, TAKEDA A, et al. Cardiac progenitor cells activated by mitochondrial delivery of resveratrol enhance the survival of a doxorubicin-induced cardiomyopathy mouse model via the mitochondrial activation of a damaged myocardium. J Controlled Release,2018, 269:177-188.
[22] YE T, ZHANG M, ZHANG Y, et al. Comparison of different rat chronic heart failure models induced by adriamycin. J Harbin Univ Com (Nat Sci Ed)(哈尔滨商业大学学报自然科学版), 2016, 32(2):154-156.
[23] LOI S, SIRTAINE N, PIETTE F, et al. Prognostic and predictive value of tumor-infiltrating lymphocytes in a phase III randomized adjuvant breast cancer trial in node-positive breast cancer comparing the addition of docetaxel to doxorubicin with doxorubicinbased chemotherapy. J Clin Oncol, 2013, 31(7):860-867.
[24] GALLUZZI L, BRAVO-SAN PEDRO J M, VITALE I, et al. Essential versus accessory aspects of cell death: recommendations of the NCCD 2015. Cell Death Differ, 2015, 22(1):58-73.
[25] JOSE CORBALAN J, VATNER D E, VATNER S F. Myocardial apoptosis in heart disease: does the emperor have clothes?. Basic Res Cardiol, 2016,111(3):31.
[26] KUZMICIC J, DEL CAMPO A, LPEZ-CRISOSTO C, et al. Mitochondrial dynamics: a potential new therapeutic target for heart failure. Rev Esp Cardiol, 2011, 64(10):916-923.
[27] BOTT-FLGEL L, WEIG H J, UHLEIN H, et al. Quantitative analysis of apoptotic markers in human end-stage heart failure. Eur J Heart Fail, 2008, 10(2):129-132.
[28] TSIPIS A, ATHANASSIADOU A M, ATHANASSIADOU P, et al. Apoptosis-related factors p53, bcl-2 and the defects of force transmission in dilated cardiomyopathy. Pathol Res Pract, 2010, 206(9):625-630.
[29] LIU H C, ZHANG Y, ZHANG S, et al. Correlation research on the protein expression (p75NTR, bax, bcl-2, and caspase-3) and cortical neuron apoptosis following mechanical injury in rat. Eur Rev Med Pharmacol Sci, 2015, 19(18):3459-3467.
[30] FRANK A K, PIETSCH E C, DUMONT P, et al. Wild-type and mutant p53 proteins interact with mitochondrial caspase-3. Cancer Biol Ther, 2011,11(8):740-745.
[31] SUBRAMANIAN S, KALYANARAMAN B, MIGRINO R Q. Mitochondrially targeted antioxidants for the treatment of cardiovascular diseases. Recent Pat Cardiovasc Drug Discov, 2010, 5(1):54-65.
[32] SHIN J W, KWON S B, BAK Y, et al. BCI induces apoptosis via generation of reactive oxygen species and activation of intrinsic mitochondrial pathway in H1299 lung cancer cells. Sci China Life Sci(中国科学), 2018,doi: 10.1007/s11427-017-9191-1.
[33] ZHANG H L, CUI S X. Reactive oxygen species and tumor therapy. J Int Oncol, 2012, 39(7):504-507.
[34] MOKWATSI G G, SCHUTTE A E, KRUGER R. A biomarker of tissue damage, lactate dehydrogenase, is associated with fibulin-1 and oxidative stress in blacks: the SAfrEIC study. Biomarkers, 2016,21(1):48-55.

基金

国家重点基础研究发展计划(973计划)项目资助(2013CB531802)
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