PMID- 34183021 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20231101 IS - 1749-8546 (Print) IS - 1749-8546 (Electronic) IS - 1749-8546 (Linking) VI - 16 IP - 1 DP - 2021 Jun 28 TI - Tanshinone I exerts cardiovascular protective effects in vivo and in vitro through inhibiting necroptosis via Akt/Nrf2 signaling pathway. PG - 48 LID - 10.1186/s13020-021-00458-7 [doi] LID - 48 AB - BACKGROUND: Tanshinone I (TI) is a primary component of Salvia miltiorrhiza Bunge (Danshen), which confers a favorable role in a variety of pharmacological activities including cardiovascular protection. However, the exact mechanism of the cardiovascular protection activity of TI remains to be illustrated. In this study, the cardiovascular protective effect and its mechanism of TI were investigated. METHODS: In this study, tert-butyl hydroperoxide (t-BHP)-stimulated H9c2 cells model was employed to investigate the protective effect in vitro. The cell viability was determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) kit. The reactive-oxygen-species (ROS) level and mitochondrial membrane potential (MMP) were investigated by the flow cytometry and JC-1 assay, respectively. While in vivo experiment, the cardiovascular protective effect of TI was determined by using myocardial ischemia-reperfusion (MI/R) model including hematoxylin-eosin (H&E) staining assay and determination of superoxide dismutase (SOD) and malondialdehyde (MDA). Tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) release were detected by Enzyme-linked immunosorbent assay (ELISA). Receptor interacting protein kinase 1 (RIP1), receptor interacting protein kinase 3 (RIP3), receptor interacting protein kinase 3 (MLKL), protein kinase B (Akt), Nuclear factor erythroid 2 related factor 2 (Nrf2), Heme oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase-1 (NQO-1) were determined by western blotting. RESULTS: Our data demonstrated that TI pretreatment attenuated t-BHP and MI/R injury-induced necroptosis by inhibiting the expression of p-RIP1, p-RIP3, and p-MLKL. TI activated the Akt/Nrf2 pathway to promote the expression of antioxidant-related proteins such as phosphorylation of Akt, nuclear factor erythroid 2 related factor 2 (Nrf2), quinone oxidoreductase-1 (NQO-1) and heme oxygenase-1 (HO-1) expression in t-BHP-stimulated H9c2 cells. TI relieved oxidative stress by mitigating ROS generation and reversing MMP loss. In vivo experiment, TI made electrocardiograph (ECG) recovery better and lessened the degree of myocardial tissue damage. The counts of white blood cell (WBC), neutrophil (Neu), lymphocyte (Lym), and the release of TNF-alpha and IL-6 were reversed by TI treatment. SOD level was increased, while MDA level was decreased by TI treatment. CONCLUSION: Collectively, our findings indicated that TI exerted cardiovascular protective activities in vitro and in vivo through suppressing RIP1/RIP3/MLKL and activating Akt/Nrf2 signaling pathways, which could be developed into a cardiovascular protective agent. FAU - Zhuo, Youqiong AU - Zhuo Y AD - College of Pharmacy, Guangxi University of Chinese Medicine, Nanning, 530000, China. AD - Guangxi Engineering Technology Research Center of Advantage Chinese Patent Drug and Ethnic Drug Development, Nanning, 530200, China. FAU - Yuan, Renyikun AU - Yuan R AD - State Key Laboratory of Innovative Drug and Efficient Energy-Saving Pharmaceutical Equipment, Jiangxi University of Traditional Chinese Medicine, Nanchang, 330004, China. FAU - Chen, Xinxin AU - Chen X AD - College of Pharmacy, Guangxi University of Chinese Medicine, Nanning, 530000, China. AD - Guangxi Engineering Technology Research Center of Advantage Chinese Patent Drug and Ethnic Drug Development, Nanning, 530200, China. FAU - He, Jia AU - He J AD - College of Pharmacy, Guangxi University of Chinese Medicine, Nanning, 530000, China. AD - Guangxi Engineering Technology Research Center of Advantage Chinese Patent Drug and Ethnic Drug Development, Nanning, 530200, China. FAU - Chen, Yangling AU - Chen Y AD - College of Pharmacy, Guangxi University of Chinese Medicine, Nanning, 530000, China. AD - Guangxi Engineering Technology Research Center of Advantage Chinese Patent Drug and Ethnic Drug Development, Nanning, 530200, China. FAU - Zhang, Chenwei AU - Zhang C AD - College of Pharmacy, Guangxi University of Chinese Medicine, Nanning, 530000, China. AD - Guangxi Engineering Technology Research Center of Advantage Chinese Patent Drug and Ethnic Drug Development, Nanning, 530200, China. FAU - Sun, Kaili AU - Sun K AD - College of Pharmacy, Guangxi University of Chinese Medicine, Nanning, 530000, China. AD - Guangxi Engineering Technology Research Center of Advantage Chinese Patent Drug and Ethnic Drug Development, Nanning, 530200, China. FAU - Yang, Shilin AU - Yang S AD - College of Pharmacy, Guangxi University of Chinese Medicine, Nanning, 530000, China. AD - Guangxi Engineering Technology Research Center of Advantage Chinese Patent Drug and Ethnic Drug Development, Nanning, 530200, China. FAU - Liu, Zhenjie AU - Liu Z AD - College of Pharmacy, Guangxi University of Chinese Medicine, Nanning, 530000, China. AD - Guangxi Engineering Technology Research Center of Advantage Chinese Patent Drug and Ethnic Drug Development, Nanning, 530200, China. FAU - Gao, Hongwei AU - Gao H AUID- ORCID: 0000-0001-9179-750X AD - College of Pharmacy, Guangxi University of Chinese Medicine, Nanning, 530000, China. gaohongwei06@126.com. AD - Guangxi Engineering Technology Research Center of Advantage Chinese Patent Drug and Ethnic Drug Development, Nanning, 530200, China. gaohongwei06@126.com. LA - eng GR - 81803807/National Natural Science Foundation of China/ PT - Journal Article DEP - 20210628 PL - England TA - Chin Med JT - Chinese medicine JID - 101265109 PMC - PMC8240219 OTO - NOTNLM OT - Akt/Nrf2 OT - Myocardial ischemia reperfusion OT - Necroptosis OT - Oxidative stress OT - RIP1/RIP3/MLKL OT - Tanshinone I COIS- The authors declare no competing conflict of interests. EDAT- 2021/06/30 06:00 MHDA- 2021/06/30 06:01 PMCR- 2021/06/28 CRDT- 2021/06/29 05:40 PHST- 2021/03/27 00:00 [received] PHST- 2021/06/16 00:00 [accepted] PHST- 2021/06/29 05:40 [entrez] PHST- 2021/06/30 06:00 [pubmed] PHST- 2021/06/30 06:01 [medline] PHST- 2021/06/28 00:00 [pmc-release] AID - 10.1186/s13020-021-00458-7 [pii] AID - 458 [pii] AID - 10.1186/s13020-021-00458-7 [doi] PST - epublish SO - Chin Med. 2021 Jun 28;16(1):48. doi: 10.1186/s13020-021-00458-7.