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Tanshinone IIA reduces apoptosis induced by hydrogen peroxide in the human endothelium-derived EA.hy926 cells.
J Ethnopharmacol. 2012 Aug 30; 143(1):100-8.JE

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Salvia Miltiorrhiza Bunge (also known as herb Danshen in Chinese) is a widely used Chinese herbal medicine. Tanshinone IIA (TSN IIA) is considered to be the most important bioactive ingredient in Danshen and exhibits an anti-atherosclerotic activity.

AIM OF STUDY

To evaluate the protective effect of TSN IIA on the human endothelial EA.hy926 cells injured by hydrogen peroxide in vitro and its possible mechanism.

MATERIALS AND METHODS

The EA.hy926 cells were incubated for 24h with different concentrations of TSN IIA (5, 10 and 20 μg/μL) or DMEM. Subsequently, cells were treated with 300 μmol/L H(2)O(2) for another 4h. Then, the percentage of cell viability was evaluated by 3-(4, 5-di-methylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. The apoptosis of EA.hy926 cells was detected by flow cytometry with AnnexinV-FITC/PI double staining and laser scanning spectral confocal technique. The generation of intracellular reactive oxygen species (ROS) generation was analyzed by flow cytometry. The mRNA expressions of caspase-3, Bcl-2 and Bax were tested by real time-reverse transcription polymerase chain reaction (real time RT-PCR). The protein expression of Bcl-2 and Bax was determined by Western blotting. MDA levels, NO production, LDH leakage, and SOD as well as caspase-3 activities were also measured using standard methods.

RESULTS

Loss of cell viability and excessive cell apoptosis were observed in EA.hy926 cells after 4h of challenge with H(2)O(2) (300 μmol/L). However, cell apoptosis was attenuated in different concentrations of TSN IIA (5, 10 and 20 μg/μL) pretreated cells. Furthermore, TSN IIA markedly inhibited the elevation of ROS evoked by H(2)O(2). Real time RT-PCR and Western blotting analysis showed that TSN IIA significantly decreased the expressions of pro-apoptotic proteins (Bax and caspase-3) while significantly increased the expression of anti-apoptotic protein Bcl-2, and resulted in obvious reduction of Bax/Bcl-2 ratio in EA.hy926 cells induced by H(2)O(2).

CONCLUSION

These observations provide preliminary evidence that TSN IIA protects EA.hy926 cells against H(2)O(2) damage, which is mainly associated with the ROS generation, followed by the imbalance of the Bax/Bcl-2 ratio, and caspase-3 activation leading to apoptosis.

Authors+Show Affiliations

Department of Biochemistry and Molecular Biology, Liaoning University of Traditional Chinese Medicine, Shenyang 110847, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22750433

Citation

Jia, Lian-Qun, et al. "Tanshinone IIA Reduces Apoptosis Induced By Hydrogen Peroxide in the Human Endothelium-derived EA.hy926 Cells." Journal of Ethnopharmacology, vol. 143, no. 1, 2012, pp. 100-8.
Jia LQ, Yang GL, Ren L, et al. Tanshinone IIA reduces apoptosis induced by hydrogen peroxide in the human endothelium-derived EA.hy926 cells. J Ethnopharmacol. 2012;143(1):100-8.
Jia, L. Q., Yang, G. L., Ren, L., Chen, W. N., Feng, J. Y., Cao, Y., Zhang, L., Li, X. T., & Lei, P. (2012). Tanshinone IIA reduces apoptosis induced by hydrogen peroxide in the human endothelium-derived EA.hy926 cells. Journal of Ethnopharmacology, 143(1), 100-8. https://doi.org/10.1016/j.jep.2012.06.007
Jia LQ, et al. Tanshinone IIA Reduces Apoptosis Induced By Hydrogen Peroxide in the Human Endothelium-derived EA.hy926 Cells. J Ethnopharmacol. 2012 Aug 30;143(1):100-8. PubMed PMID: 22750433.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Tanshinone IIA reduces apoptosis induced by hydrogen peroxide in the human endothelium-derived EA.hy926 cells. AU - Jia,Lian-Qun, AU - Yang,Guan-Lin, AU - Ren,Lu, AU - Chen,Wen-Na, AU - Feng,Jun-Yi, AU - Cao,Yang, AU - Zhang,Lin, AU - Li,Xue-Tao, AU - Lei,Ping, Y1 - 2012/06/28/ PY - 2012/02/13/received PY - 2012/05/16/revised PY - 2012/06/06/accepted PY - 2012/7/4/entrez PY - 2012/7/4/pubmed PY - 2012/12/19/medline SP - 100 EP - 8 JF - Journal of ethnopharmacology JO - J Ethnopharmacol VL - 143 IS - 1 N2 - ETHNOPHARMACOLOGICAL RELEVANCE: Salvia Miltiorrhiza Bunge (also known as herb Danshen in Chinese) is a widely used Chinese herbal medicine. Tanshinone IIA (TSN IIA) is considered to be the most important bioactive ingredient in Danshen and exhibits an anti-atherosclerotic activity. AIM OF STUDY: To evaluate the protective effect of TSN IIA on the human endothelial EA.hy926 cells injured by hydrogen peroxide in vitro and its possible mechanism. MATERIALS AND METHODS: The EA.hy926 cells were incubated for 24h with different concentrations of TSN IIA (5, 10 and 20 μg/μL) or DMEM. Subsequently, cells were treated with 300 μmol/L H(2)O(2) for another 4h. Then, the percentage of cell viability was evaluated by 3-(4, 5-di-methylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. The apoptosis of EA.hy926 cells was detected by flow cytometry with AnnexinV-FITC/PI double staining and laser scanning spectral confocal technique. The generation of intracellular reactive oxygen species (ROS) generation was analyzed by flow cytometry. The mRNA expressions of caspase-3, Bcl-2 and Bax were tested by real time-reverse transcription polymerase chain reaction (real time RT-PCR). The protein expression of Bcl-2 and Bax was determined by Western blotting. MDA levels, NO production, LDH leakage, and SOD as well as caspase-3 activities were also measured using standard methods. RESULTS: Loss of cell viability and excessive cell apoptosis were observed in EA.hy926 cells after 4h of challenge with H(2)O(2) (300 μmol/L). However, cell apoptosis was attenuated in different concentrations of TSN IIA (5, 10 and 20 μg/μL) pretreated cells. Furthermore, TSN IIA markedly inhibited the elevation of ROS evoked by H(2)O(2). Real time RT-PCR and Western blotting analysis showed that TSN IIA significantly decreased the expressions of pro-apoptotic proteins (Bax and caspase-3) while significantly increased the expression of anti-apoptotic protein Bcl-2, and resulted in obvious reduction of Bax/Bcl-2 ratio in EA.hy926 cells induced by H(2)O(2). CONCLUSION: These observations provide preliminary evidence that TSN IIA protects EA.hy926 cells against H(2)O(2) damage, which is mainly associated with the ROS generation, followed by the imbalance of the Bax/Bcl-2 ratio, and caspase-3 activation leading to apoptosis. SN - 1872-7573 UR - https://www.unboundmedicine.com/medline/citation/22750433/Tanshinone_IIA_reduces_apoptosis_induced_by_hydrogen_peroxide_in_the_human_endothelium_derived_EA_hy926_cells_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0378-8741(12)00394-7 DB - PRIME DP - Unbound Medicine ER -