Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (9): 1014-1021.doi: 10.3736/jcim20110913

• Original Experimental Research • Previous Articles     Next Articles

Scutellarin benzyl ester partially secured the ischemic injury by its anti-apoptosis mechanism in cardiomyocytes of neonatal rats

Hao Dai1, Jun Gu1,2, Ling-zhi Li1,2, Li-mei Yang1, Hui Liu3, Jian-yu Li1,2()   

  1. 1. Division of Pharmaceutical Chemistry,Medical College of Chinese People’s Armed Police Forces,Tianjin 300162, China;
    2. Tianjin Key Laboratory of Occupational and Environmental Hazards Biomarkers, Tianjin 300162, China
    3. Institute of Hepatobiliary Diseases, Third Central Hospital of Tianjin Medical University, Tianjin 300170, China
  • Received:2011-04-14 Accepted:2011-05-27 Online:2011-09-20 Published:2011-09-15
  • Contact: Li Jian-yu

Objective: To investigate the protective effects of scutellarin benzyl ester on neonatal rats’ cardiomyocytes injured by ischemia and its anti-apoptosis mechanism.

Methods: The cardiomyocytes in primary culture were prepared from ventricular tissue of 1- to 3-day-old Sprague-Dawley rats and the cells in good condition were assigned to five groups: control group, ischemic model group and three scutellarin benzyl ester groups (doses of 100, 50 and 25 μmol/L, respectively). The model of ischemic injury was established in the primary culture of cardiomyocytes under glucose-free anoxic condition. After ischemia for 6 h, the metabolic ability of the cells was detected by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assay and the activity of lactate dehydrogenase (LDH) in the media was measured by biochemistry approaches. The nuclear damage was revealed by Hoechst-propidium iodide staining. The percentage of apoptotic cells was monitored by flow cytometry. The expression levels of cytochrome c and caspase-3 mRNAs and proteins were determined by reverse transcription-polymerase chain reaction and Western blotting, respectively.

Results: After exposure to ischemic condition, the cell viability of the model group was degraded compared with that of the control group (P<0.01) and scutellarin benzyl ester (high and medium doses) could attenuate the loss of cell viability induced by ischemia (P<0.01 and P<0.05). In addition, each dose of scutellarin benzyl ester could significantly reduce the release of LDH from cardiomyocytes injured by ischemia (P<0.01). In morphology, the injured nuclei presented significant changes such as condensation of chromatin, and shrinkage and fragmentation of nuclei, which could be attenuated remarkably by pretreatment with scutellarin benzyl ester. Furthermore, scutellarin benzyl ester could significantly decrease the percentage of apoptosis induced by ischemia (P<0.01) and inhibit the increased expression levels of cytochrome c and caspase-3 mRNAs and proteins (P<0.01).

Conclusion: Scutellarin benzyl ester has a remarkable protective effect against myocardial ischemic injury and the protective mechanism may associate with its anti-apoptosis effect by inhibiting cytochrome c release and caspase-3 activation.

Key words: Erigeron breviscapus, scutellarin benzyl ester, prodrugs, myocytes, cardiac, ischemia, apoptosis, rats


Group n OD value
Control 6 0.517±0.054
Model 6 0.215±0.025**
High-dose scutellarin benzyl ester
(100 μmol/L)
6 0.287±0.023**△△
Medium-dose scutellarin benzyl ester
(50 μmol/L)
6 0.253±0.023**△
Low-dose scutellarin benzyl ester
(25 μmol/L)
6 0.210±0.021**


Group n LDH
Control 6 78.8±12.9
Model 6 288.1±30.7**
High-dose scutellarin benzyl ester
(100 μmol/L)
6 64.2±22.9△△
Medium-dose scutellarin benzyl ester
(50 μmol/L)
6 101.4±22.7△△
Low-dose scutellarin benzyl ester
(25 μmol/L)
6 141.3±23.1**△△

Figure 1

Changes of nuclei tested by Hoechst-prodium iodide staining (Microscopy, ×400)A: Control; B: Model; C: Scutellarin benzyl ester (100 μmol/L)."

Figure 2

Changes of apoptosis percentage tested by flow cytometryA: Control group; B: Model group; C: Scutellarin benzyl ester (100 μmol/L) group."


Group n Apoptosis percentage
Control 6 4.52±0.29
Model 6 30.14±1.89**
High-dose scutellarin benzyl ester
(100 μmol/L)
6 18.93±1.45**△△
Medium-dose scutellarin benzyl ester
(50 μmol/L)
6 22.10±1.27**△△
Low-dose scutellarin benzyl ester
(25 μmol/L)
6 29.89±2.12**△△

Figure 3

Changes in the expression of cytochrome c mRNA tested by RT-PCRA: Control group; B: Model group; C: Scutellarin benzyl ester (100 μmol/L) group; D: Scutellarin benzyl ester (50 μmol/L) group; E: Scutellarin benzyl ester (25 μmol/L) group. **P<0.01, vs control group; △△P<0.01, vs model group. RT-PCR: reverse transcription-polymerase chain reaction; GAPDH: glyceraldehyde-3-phosphate dehydrogenase."

Figure 4

Changes in the expression of caspase-3 mRNA tested by RT-PCRA: Control group; B: Model group; C: Scutellarin benzyl ester (100 μmol/L) group; D: Scutellarin benzyl ester (50 μmol/L) group; E: Scutellarin benzyl ester (25 μmol/L) group. **P<0.01, vs control group; △△P<0.01, vs model group. RT-PCR: reverse transcription-polymerase chain reaction; GAPDH: glyceraldehyde-3-phosphate dehydrogenase."

Figure 5

Changes in the expression of cytochrome c protein tested by Western blotting A: Control group; B: Model group; C: Scutellarin benzyl ester (100 μmol/L) group; D: Scutellarin benzyl ester (50 μmol/L) group; E: Scutellarin benzyl ester (25 μmol/L) group. **P<0.01, vs control group; △△P<0.01, vs model group."

Figure 6

Changes in the expression of caspase-3 protein tested by Western blotting A: Control group; B: Model group; C: Scutellarin benzyl ester (100 μmol/L) group; D: Scutellarin benzyl ester (50 μmol/L) group; E: Scutellarin benzyl ester (25 μmol/L) group. **P<0.01, vs control group; △△P<0.01, vs model group."

[1] Liu H, Yang X, Tang R, Liu J, Xu H . Effect of scutellarin on nitric oxide production in early stages of neuron damage induced by hydrogen peroxide[J]. Pharmacol Res, 2005,51(3):205-210
doi: 10.1016/j.phrs.2004.09.001
[2] Lu J, Cheng C, Zhao X, Liu Q, Yang P, Wang Y, Luo G . PEG-scutellarin prodrugs: synjournal, water solubility and protective effect on cerebral ischemia/reperfusion injury[J]. Eur J Med Chem, 2010,45(5):1731-1738
doi: 10.1016/j.ejmech.2010.01.006
[3] Gao ZX, Huang DY, Li HX, Zhang LN, Lv YH, Cui HD, Zheng JH . Scutellarin promotes in vitro angiogenesis in human umbilical vein endothelial cells[J]. Biochem Biophys Res Commun, 2010,400(1):151-156
doi: 10.1016/j.bbrc.2010.08.034 pmid: 20709020
[4] van Gelder J, Shafiee M, De Clercq E, Penninckx F, van den Mooter G, Kinget R, Augustijns P . Species-dependent and site-specific intestinal metabolism of ester prodrugs[J]. Int J Pharm, 2000,205(1-2):93-100
doi: 10.1016/S0378-5173(00)00507-X pmid: 11000545
[5] Yang LM, Gu J, Li JY, Dai H, Li LZ . Synjournal of scutellarin derivatives and study of their anti-heart ischemia activities in rats[J]. Tianjin Zhong Yi Yao, 2010,27(3):240-242
杨丽梅, 顾军, 李建宇, 代皓, 李灵芝 . 灯盏乙素衍生物的合成及抗大鼠心肌缺血活性研究[J]. 天津中医药, 2010,27(3):240-242
[6] Cao F, Guo JX, Ping QN, Liao ZG . Prodrugs of scutellarin: ethyl, benzyl and N,N-diethylglycolamide ester synjournal, physicochemical properties, intestinal metabolism and oral bioavailability in the rats[J]. Eur J Pharm Sci, 2006,29(5):385-393
doi: 10.1016/j.ejps.2006.07.007
[7] Brunotto M, Malberti A, Zarate AM, Barra JL, Calderón O, Pinas E, Plavnik L, Crosa M . Early phenotypic and genotypic alterations in submandibular gland oncogenesis in rats[J]. Acta Odontol Latinoam, 2006,19(1):13-21
pmid: 17121194
[8] Li JY, Li LZ, Gong HY, Li Y, Zhang XN . Effect of Potentilla anserina L. on cardiomyocyte apoptosis induced by hypoxia[J]. Zhonghua Lao Nian Xin Nao Xue Guan Bing Za Zhi, 2007,9(9):619-622
doi: 10.3969/j.issn.1009-0126.2007.09.015
李建宇, 李灵芝, 龚海英, 李怡, 张新宁 . 蕨麻对缺氧诱导心肌细胞凋亡的影响[J]. 中华老年心脑血管病杂志, 2007,9(9):619-622
doi: 10.3969/j.issn.1009-0126.2007.09.015
[9] Li JY, Li LZ, Zhang YL, Zhang XN, Gong HY . Protective effect of Potentilla anserina L. on hypoxia injury in neonatal rats cardiomyocytes[J]. Zhongguo Xin Yao Za Zhi, 2007,16(12):944-946
李建宇, 李灵芝, 张永亮, 张新宁, 龚海英 . 蕨麻醇提物对心肌细胞缺氧损伤的保护作用[J]. 中国新药杂志, 2007,16(12):944-946
[10] Li JY, Li Y, Gong HY, Zhao XB, Li LZ . Protective effects of n-butanol extract of Potentilla anserina on acute myocardial ischemic injury in mice[J]. J Chin Integr Med, 2009,7(1):48-52
李建宇, 李怡, 龚海英, 赵晓博, 李灵芝 . 蕨麻正丁醇提取部位对小鼠急性心肌缺血损伤的保护作用[J]. 中西医结合学报, 2009,7(1):48-52
[11] Tang YH, Zhang SP, Liang Y, Deng CQ . Effects of Panax notoginseng saponins on mRNA expressions of interleukin-1β, its correlative factors and cysteinyl-aspartate specific protease after cerebral ischemia-reperfusion in rats[J]. J Chin Integr Med, 2007,5(3):328-332
doi: 10.3969/j.issn.1005-9903.2010.16.038
唐映红, 张淑萍, 梁燕, 邓常清 . 三七总皂苷对大鼠脑缺血再灌注后白细胞介素-1β和其相关因子及半胱氨酸天冬氨酸特异性蛋白酶表达的影响[J]. 中西医结合学报, 2007,5(3):328-332
doi: 10.3969/j.issn.1005-9903.2010.16.038
[12] Rong F, Peng Z, Ye MX, Zhang QY, Zhao Y, Zhang SM, Guo HT, Hui B , Wang infarction after ischemia/reperfusion are attenuated by κ-opioid receptor agonist[J]. Arch Med Res, 2009,40(4):227-234
doi: 10.1016/j.arcmed.2009.04.009
[13] Sodha NR, Clements RT, Feng J, Liu Y, Bianchi C, Horvath EM, Szabo C, Sellke FW . The effects of therapeutic sulfide on myocardial apoptosis in response to ischemia-reperfusion injury[J]. Eur J Cardiothorac Surg, 2008,33(5):906-913
doi: 10.1016/j.ejcts.2008.01.047
[14] Celhar T, Batista Napotnik T, Obreza A, Zega A, Anderluh PS, Kikelj D, Mlinaric-Rascan I . Azaphenylalanine-based serine protease inhibitors induce caspase-mediated apoptosis[J]. Eur J Pharmacol, 2009,602(1):15-22
doi: 10.1016/j.ejphar.2008.11.008 pmid: 19032950
[15] Su BH, Chiu HY, Soga T, Lin KJ, Hsu CT . Ulinastatin alone does not reduce caspase 3-mediated apoptosis in protease-positive Aeromonas hydrophilia-induced sepsis[J]. J Formos Med Assoc, 2007,106(2):97-104
doi: 10.1016/S0929-6646(09)60224-2
[1] Cai-lian Fana, Wan-jun Caib, Meng-nan Ye, Miao Chen, Yi Dai. Qili Qiangxin, a compound herbal medicine formula, alleviates hypoxia-reoxygenation-induced apoptotic and autophagic cell death via suppression of ROS/AMPK/mTOR pathway in vitro. Journal of Integrative Medicine, 2022, 20(4): 365-375.
[2] Hong-xiao Li, Ling Shi, Shang-jie Liang Chen-chen Fang, Qian-qian Xu, Ge Lu, Qian Wang, Jie Cheng, Jie Shen, Mei-hong Shen. Moxibustion alleviates decreased ovarian reserve in rats by restoring the PI3K/AKT signaling pathway. Journal of Integrative Medicine, 2022, 20(2): 163-172.
[3] Mohamed S. Othman, Sofian T. Obeidat, Amal H. Al-Bagawi, Mohamed A. Fareid, Alaa Fehaid, Ahmed E. Abdel Moneim. Green-synthetized selenium nanoparticles using berberine as a promising anticancer agent#br#
. Journal of Integrative Medicine, 2022, 20(1): 65-72.
[4] Hyongjun Jeon, Hee-Young Kim, Chang-Hwan Bae, Yukyung Lee, Sungtae Koo, Seungtae Kim. Korean red ginseng decreases 1-methyl-4-phenylpyridinium-induced mitophagy in SH-SY5Y cells. Journal of Integrative Medicine, 2021, 19(6): 537-544.
[5] Bing-rong Li, Shi-yun Shao, Long Yuan, Ru Jia, Jian Sun, Qing Ji, Hua Sui, Li-hong Zhou, Yi Zhang, Hui Liu, Qi Li, Yan Wang, Bi-meng Zhang. Effects of mild moxibustion on intestinal microbiome and NLRP3 inflammasome in rats with 5-fluorouracil-induced intestinal mucositis. Journal of Integrative Medicine, 2021, 19(2): 144-157.
[6] Porwornwisit Tritripmongkol, Tullayakorn Plengsuriyakarn, Mayuri Tarasuk, Kesara Na-Bangchang. In vitro cytotoxic and toxicological activities of ethanolic extract of Kaempferia galanga Linn. and its active component, ethyl-p-methoxycinnamate, against cholangiocarcinoma. Journal of Integrative Medicine, 2020, 18(4): 326-333.
[7] Sitthichai Iamsaard, Supatcharee Arun, Jaturon Burawat, Supataechasit Yannasithinon, Saranya Tongpan, Sudtida Bunsueb, Natthapol Lapyuneyong, Pannawat Choowong-in, Nareelak Tangsrisakda, Chadaporn Chaimontri, Wannisa Sukhorum. Evaluation of antioxidant capacity and reproductive toxicity of aqueous extract of Thai Mucuna pruriens seeds. Journal of Integrative Medicine, 2020, 18(3): 265-273.
[8] Varuni Colamba Pathiranage, Ira Thabrew, Sameera R. Samarakoon, Kamani H. Tennekoon, Umapriyatharshini Rajagopalan, Meran K. Ediriweera. Evaluation of anticancer effects of a pharmaceutically viable extract of a traditional polyherbal mixture against non-small-cell lung cancer cells. Journal of Integrative Medicine, 2020, 18(3): 242-252.
[9] Maged Mohamed Maher Abou-Hashem, Dina Mohamed Abo-elmatty, Noha Mostafa Mesbah, Ahmed Mohamed Abd EL-Mawgoud. Induction of sub-G0 arrest and apoptosis by seed extract of Moringa peregrina (Forssk.) Fiori in cervical and prostate cancer cell lines. Journal of Integrative Medicine, 2019, 17(6): 410-422.
[10] Supakanya Kumkarnjana, Rutt Suttisri, Ubonthip Nimmannit, Apirada Sucontphunt, Mattaka Khongkow, Thongchai Koobkokkruad, Nontima Vardhanabhuti. Flavonoids kaempferide and 4,2′-dihydroxy-4′,5′,6′-trimethoxychalcone inhibit mitotic clonal expansion and induce apoptosis during the early phase of adipogenesis in 3T3-L1 cells. Journal of Integrative Medicine, 2019, 17(4): 288-295.
[11] Shriniwas S. Basaiyye, Sanjay Kashyap, Kannan Krishnamurthi, Saravanadevi Sivanesan. Induction of apoptosis in leukemic cells by the alkaloid extract of garden cress (Lepidium sativum L.). Journal of Integrative Medicine, 2019, 17(3): 221-228.
[12] Carlos R. Oliveira, Daniel G. Spindola, Daniel M. Garcia, Adolfo Erustes, Alexandre Bechara, Caroline Palmeira-dos-Santos, Soraya S. Smaili, Gustavo J.S. Pereira, André Hinsberger, Ezequiel P. Viriato, Maria Cristina Marcucci, Alexandra C.H.F. Sawaya, Samantha L. Tomaz, Elaine G. Rodrigues, Claudia Bincoletto. Medicinal properties of Angelica archangelica root extract: Cytotoxicity in breast cancer cells and its protective effects against in vivo tumor development. Journal of Integrative Medicine, 2019, 17(2): 132-140.
[13] Bruno José Martins Da Silva, Sandro Wilson Gomes Pereira, Ana Paula Drummond Rodrigues, José Luiz Martins Do Nascimento, Edilene Oliveira Silva. In vitro antileishmanial effects of Physalis angulata root extract on Leishmania infantum. Journal of Integrative Medicine, 2018, 16(6): 404-410.
[14] Morufu Eyitayo Balogun, Elizabeth Enohnyaket Besong, Jacinta Nkechi Obimma, Ogochukwu Sophia Mbamalu, Fankou Serges Athanase Djobissie. Protective roles of Vigna subterranea (Bambara nut) in rats with aspirin-induced gastric mucosal injury. Journal of Integrative Medicine, 2018, 16(5): 342-349.
[15] Lucky Legbosi Nwidu, Raphael Ellis Teme. Hot aqueous leaf extract of Lasianthera africana (Icacinaceae) attenuates rifampicin-isoniazid-induced hepatotoxicity. Journal of Integrative Medicine, 2018, 16(4): 263-272.
Full text



[1] Jin-rong Fu. Establishment of multivariate diagnosis and treatment system of modern gynecology of traditional Chinese medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 22-24
[2] Hao Li, Ming-jiang Yao, Wen-ming Zhao, Jie Guan, Lin-lin Cai, Ling Cui. A randomized, controlled, double-blind trial of Huannao Yicong capsule in senile patients with mild cognitive impairment. Journal of Chinese Integrative Medicine, 2008, 6(1): 25-31
[3] Zhi-chun Jin. Problems in establishing clinical guideline for integrated traditional Chinese and Western medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 5-8
[4] SUN Ming-yu, ZUO Jian, DUAN Ji-feng, HAN Jun, FAN Shi-ming, ZHANG Wei, ZHU Li-fang, YAO Ming-hui. Antitumor activities of kushen flavonoids in vivo and in vitro. Journal of Chinese Integrative Medicine, 2008, 6(1): 51-59
[5] Min Cheng, Qiong Feng, Shu-wen Qian, Hui Gao, Cui-qing Zhu. Preliminary assay of p-amyloid binding elements in heart-beneficial recipe. Journal of Chinese Integrative Medicine, 2008, 6(1): 68-72
[6] Ning-qun Wang, Liang-duo Jiang, Zong-xing Li. Research progress in asthma-related quality of life. Journal of Chinese Integrative Medicine, 2008, 6(1): 93-97
[7] Jing-yuan Mao, Chang-xiao Liu, Heng-he Wang, Guang-li Wei , Zhen-peng Zhang, Jie Xing, Wang Xian liang , Ying-fei Bi . Effects of Shenmai Injection on serum concentration and pharmacokinetics of digoxin in dogs with heart failure. Journal of Chinese Integrative Medicine, 2010, 8(11): 1070-1074
[8] Zhi-mei Wang, Bin Zhang. A study on translation of ellipses in Huangdi Neijing from perspective of hermeneutic theory. Journal of Chinese Integrative Medicine, 2010, 8(11): 1097-1100
[9] Gui Yu, Jie Wang. Thinking on building the network cardiovasology of Chinese medicine. Journal of Chinese Integrative Medicine, 2012, 10(11): 1206-1210
[10] Pedro Saganha João, Doenitz Christoph, Greten Tobias, Efferth Thomas, J. Greten Henry. Qigong therapy for physiotherapists suffering from burnout: a preliminary study. Journal of Chinese Integrative Medicine, 2012, 10(11): 1233-1239