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1Traditional Medicine Research  2018, Vol. 3 Issue (3): 140-147    DOI: 10.12032/TMR201811072
Modernization of Traditional Medicine     
Polysaccharide extracts of Cirsium japonicum protect rat H9c2 myocardial cells from oxidative stress induced by hydrogen peroxide
Zheng-Bo Tao1, Li-Yan Xiong2, Li-Hui Wang3,*(), Chuan Zhang2,*()
1 Administrative Office for Undergraduates, Second Military Medical University, Shanghai, China.
2 Department of Identification of Traditional Chinese Medicine, School of Pharmacy, Second Military Medical University, Shanghai, China.
3 Business Unit of Traditional Chinese Medicine Resources, Shanghai Traditional Chinese Medicine Co., Ltd, Shanghai Pharma, Shanghai, China.
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Highlights

The current study found that the polysaccharide extract of Daji (Cirsium japonicum) have a favorable therapeutic effect against oxidative stress and the underlying mechanism might be involved with MAPK pathway.

Editor’s Summary

Daji (Cirsium japonicum) have been traditionally utilized by the Tibetan and Mongolian people since the Tang Dynasty of China (618 A.D. - 907 A.D.).

Abstract

Daji (Cirsium japonicum) has been applied against gastric disorders, lung diseases, and cardiovascular problems in the traditional Chinese medicinal system. The present study was to investigate the protective effects of Daji (Cirsium japonicum) polysaccharide extracts (CJP) against hydrogen peroxide (H2O2) shock in rat H9c2 myocardial cells. First, CJP was isolated by hot water extraction and ethanol precipitation; it was then characterized by high performance liquid chromatography and infrared spectrum analysis. Rat H9c2 cells were subjected to H2O2 treatment to establish a cell injury model. The 3- (4,5- dimethylthiazol- 2-yl)-2,5- diphenyltetrazolium bromide assay showed that CJP pretreatment significantly ameliorated the H2O2 injury in a dose-dependent manner. Furthermore, the cell apoptosis induced by H2O2 was markedly inhibited by CJP pretreatment, whereas the cleavage level of caspase-3, -8, and -9 was reduced. In addition, the p38 mitogen-activated protein kinase pathway might be involved in the protective effect of CJP on myocardial cells. Therefore, we conclude that polysaccharide extracts of Daji (Cirsium japonicum) protect rat H9c2 myocardial cells from oxidative stress induced by H2O2.



Key wordsDaji (Cirsium japonicum)      Polysaccharide      Hydrogen peroxide (H2O2)      H9c2 myocardial cell      Apoptosis     
Published: 04 May 2018
Fund:  Funding: This work was supported by Shanghai Key Discipline Construction Project of Clinical Pharmacology (2016).
Corresponding Authors: Wang Li-Hui,Zhang Chuan     E-mail: wanglihui.yh@qq.com.;zhangchuan@smmu.edu.cn.
About author: #These authors contributed equally to this work.
Cite this article:

Zheng-Bo Tao, Li-Yan Xiong, Li-Hui Wang, Chuan Zhang. Polysaccharide extracts of Cirsium japonicum protect rat H9c2 myocardial cells from oxidative stress induced by hydrogen peroxide. 1Traditional Medicine Research, 2018, 3(3): 140-147. doi: 10.12032/TMR201811072

URL:

https://www.tmrjournals.com/tmr/EN/10.12032/TMR201811072     OR     https://www.tmrjournals.com/tmr/EN/Y2018/V3/I3/140

Figure 1 Phytochemical analysis of CJP
(A) High-performance gel permeation chromatography of CJP, analyzed on a TSK GMPWXL gel filtration column (7.6 mm×300 mm, TOSOH), and eluted with ultrapure water at a flow rate of 1.0 ml/min, and detected at 254 nm. (B) Fourier transform of infrared spectra for CJP. CJP, Daji (Cirsium japonicum) polysaccharide extracts.
Figure 2 CJP significantly mitigated the H2O2-induced injury through inhibition of apoptosis
(A) The cell viability level of H9c2 cells treated by H2O2 (25, 50, and 100 mol/L) or CJP (50, 100, 200, 400, and 600 μg/mL). (B) CJP with a concentration of no less than 100μg/mL exerted protective effect on H2c9 cells treated by H2O2. (##P < 0.001, H2O2 vs Con; *P = 0.045 , H2O2 + CJP100 vs H2O2; *P = 0.010, H2O2 + CJP200 vs H2O2; **P < 0.001, H2O2 + CJP400 vs H2O2; **P = 0.001, H2O2 + CJP600 vs H2O2.). (C) Representative ?uorescence images showed that CJP pre-treatment significantly ameliorated the morphological alteration of H9c2 cell nuclei induced by H2O2 (Magnification, 200×). (D) Representative flow cytometric plots demonstrated that CJP pretreatment significantly inhibited the cell apoptosis in H2c9 cells induced by H2O2. (E) Typical Western blots showed activation of caspase-3, -8, and -9 was significantly inhibited by CJP in H2c9 cells treated by H2O2. Values are means ± SEM, n = 6 per group. CJP, Daji (Cirsium japonicum) polysaccharide extracts; H2O2, Hydrogen peroxide.
Figure 3 CJP significantly inhibited the phosphorylation of p38 and JNK MAPK pathway in H9c2 cardiomyocytes treated by H2O2
(A) Typical blots; (B) Densitometric analysis of western blot of p-JNK/JNK. (##P < 0.001, H2O2 vs Con; **P = 0.005, H2O2 + CJP100 vs H2O2.) (C) Densitometric analysis of western blot of p-p38/p38. (##P = 0.001, H2O2 vs Con; **P = 0.006, H2O2 + CJP100 vs H2O2.) Values are means ± SEM, n = 6 per group.
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