Triptolide reduces ischemia/reperfusion injury in rats and H9C2 cells via inhibition of NF‑κB, ROS and the ERK1/2 pathway

Yang,Bin; Yan,Ping; Yang,Guang‑Zhao; Cao,Hui‑Li; Wang,Fei; Li,Bao

doi:10.3892/ijmm.2018.3537

Triptolide reduces ischemia/reperfusion injury in rats and H9C2 cells via inhibition of NF‑κB, ROS and the ERK1/2 pathway

Authors:
- Bin Yang
- Ping Yan
- Guang‑Zhao Yang
- Hui‑Li Cao
- Fei Wang
- Bao Li
View Affiliations

Affiliations: Department of Cardiovascular Medicine, The Second Affiliated Hospital of Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China, Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China, Department of Cardiovascular Medicine, The First Affiliated Hospital of Shanxi Medical University, Taiyuan, Shanxi 030012, P.R. China, Department of Cardiovascular Medicine, Shanxi Cardiovascular Hospital, Taiyuan, Shanxi 030024, P.R. China, Department of Cardiovascular Medicine, The Second Affiliated Hospital of Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China
Published online on: March 6, 2018 https://doi.org/10.3892/ijmm.2018.3537
Pages: 3127-3136
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Myocardial ischemia/reperfusion (I/R) induces cardiac cell injury; however, the mechanism underlying cardiac damage remains unclear. A previous study demonstrated that triptolide (TP) exerts protective effects against I/R in cerebral cells. The present study aimed to evaluate the protective effects of TP on cardiac cells, and investigated the potential mechanisms involved in I/R‑induced damage. Rats and cardiac H9C2 cells undergoing I/R were pretreated with TP, and cell damage was assessed in vivo and in vitro. Hematoxylin and eosin and terminal deoxynucleotidyl‑transferase‑mediated dUTP nick end labeling staining were employed to evaluate I/R injury in rat cardiac tissue. Inflammatory factors, including tumor necrosis factor‑α, interleukin (IL)‑1β and IL‑6, were detected by ELISA. Biochemical analyses were performed to evaluate the bioactivity of superoxide dismutase, malondialdehyde and catalase. In addition, viability of H9C2 cells was measured using the Cell Counting kit 8 assay. Flow cytometry was used to evaluate cell apoptosis and reactive oxygen species (ROS) generation. Furthermore, the expression levels of proteins associated with apoptosis, peroxide and inflammation were measured using western blot analysis. H9C2 cells were also treated with N‑acetylcysteine and pyrrolidine dithiocarbamate, and cell injury was assessed after peroxidation or I/R. The results demonstrated that TP exerted a significant protective effect on cardiac cells in vivo and in vitro. TP reduced the inflammatory response, as determined by nuclear factor‑κB inhibition. In addition, TP decreased ROS‑mediated lipid peroxidation, and reduced ROS generation. TP also inhibited cell apoptosis by activating the extracellular signal‑regulated kinase 1/2 pathway. In conclusion, TP may protect cardiac cells from I/R injury; the potential protective mechanisms of TP against I/R include anti‑inflammatory action, antioxidation and apoptotic resistance.

View Figures

View References

1	Durukan A and Tatlisumak T: Acute ischemic stroke: Overview of major experimental rodent models, pathophysiology, and therapy of focal cerebral ischemia. Pharmacol Biochem Behav. 87:179–197. 2007. View Article : Google Scholar : PubMed/NCBI
2	Donnan GA, Fisher M, Macleod M and Davis SM: Stroke. Lancet. 71:1612–1623. 2008. View Article : Google Scholar
3	Molina CA and Alvarez-Sabín J: Recanalization and reperfusion therapies for acute ischemic stroke. Cerebrovasc Dis. 27(Suppl 1): 162–167. 2009. View Article : Google Scholar : PubMed/NCBI
4	Peralta C, Bulbena O, Xaus C, Prats N, Cutrin JC, Poli G, Gelpi E and Roselló-Catafau J: Ischemic preconditioning: A defense mechanism against the reactive oxygen species generated after hepatic ischemia reperfusion. Transplantation. 73:1203–1211. 2002. View Article : Google Scholar : PubMed/NCBI
5	Eltzschig HK and Eckle T: Ischemia and reperfusion-from mechanism to translation. Nat Med. 17:1391–1401. 2011. View Article : Google Scholar : PubMed/NCBI
6	Huang J, Upadhyay UM and Tamargo RJ: Inflammation in stroke and focal cerebral ischemia. Surg Neurol. 66:232–245. 2006. View Article : Google Scholar : PubMed/NCBI
7	Esterbauer H and Cheeseman KH: Determination of aldehydic lipid peroxidation products: Malonaldehyde and 4-hydroxynonenal. Methods Enzymol. 186:407–421. 1990. View Article : Google Scholar : PubMed/NCBI
8	Tuttolomondo A, Di Sciacca R, Di Raimondo D, Renda C, Pinto A and Licata G: Inflammation as a therapeutic target in acute ischemic stroke treatment. Curr Top Med Chem. 9:1240–1260. 2009. View Article : Google Scholar : PubMed/NCBI
9	Faul JL, Nishimura T, Berry GJ, Benson GV, Pearl RG and Kao PN: Triptolide attenuates pulmonary arterial hypertension and neointimal formation in rats. Am J Respir Crit Care Med. 162:2252–2258. 2000. View Article : Google Scholar : PubMed/NCBI
10	Hao M, Li X, Feng J and Pan N: Triptolide protects against ischemic stroke in rats. Inflammation. 38:1617–1623. 2015. View Article : Google Scholar : PubMed/NCBI
11	Hoyle GW, Hoyle CI, Chen J, Chang W, Williams RW and Rando RJ: Identification of triptolide, a natural diterpenoid compound, as an inhibitor of lung inflammation. Am J Physiol Lung Cell Mol Physiol. 298:L830–L836. 2010. View Article : Google Scholar : PubMed/NCBI
12	Wu C, Xia Y, Wang P, Lu L and Zhang F: Triptolide protects mice from ischemia/reperfusion injury by inhibition of IL-17 production. Int Immunopharmacol. 11:1564–1572. 2011. View Article : Google Scholar : PubMed/NCBI
13	He JK, Yu SD, Zhu HJ, Wu JC and Qin ZH: Triptolide inhibits NF-kappaB activation and reduces injury of donor lung induced by ischemia/reperfusion. Acta Pharmacol Sin. 28:1919–1923. 2007. View Article : Google Scholar : PubMed/NCBI
14	Lin N, Liu C, Xiao C, Jia H, Imada K, Wu H and Ito A: Triptolide, a diterpenoid triepoxide, suppresses inflammation and cartilage destruction in collagen-induced arthritis mice. Biochem Pharmacol. 73:136–146. 2007. View Article : Google Scholar
15	Lu Y, Liu Y, Fukuda K, Nakamura Y, Kumagai N and Nishida T: Inhibition by triptolide of chemokine, proinflammatory cytokine, and adhesion molecule expression induced by lipopolysaccharide in corneal fibroblasts. Invest Ophthalmol Vis Sci. 47:3796–3800. 2006. View Article : Google Scholar : PubMed/NCBI
16	Matta R, Wang X, Ge H, Ray W, Nelin LD and Liu Y: Triptolide induces anti-inflammatory cellular responses. Am J Transl Res. 1:267–282. 2009.PubMed/NCBI
17	Chandel NS, Trzyna WC, McClintock DS and Schumacker PT: Role of oxidants in NF-kappa B activation and TNF-alpha gene transcription induced by hypoxia and endotoxin. J Immunol. 165:1013–1021. 2000. View Article : Google Scholar : PubMed/NCBI
18	Lakhan SE, Kirchgessner A and Hofer M: Inflammatory mechanisms in ischemic stroke: Therapeutic approaches. J Transl Med. 7:972009. View Article : Google Scholar : PubMed/NCBI
19	Jin XQ, Ye F, Zhang JJ, Zhao Y and Zhou XL: Triptolide attenuates cerebral ischemia and reperfusion injury in rats through the inhibition the nuclear factor kappa B signaling pathway. Neuropsychiatr Dis Treat. 11:1395–1403. 2015.PubMed/NCBI
20	Hur GM, Ryu YS, Yun HY, Jeon BH, Kim YM, Seok JH and Lee JH: Hepatic ischemia/reperfusion in rats induces iNOS gene transcription by activation of NF-kappaB. Biochem Biophys Res Commun. 261:917–922. 1999. View Article : Google Scholar : PubMed/NCBI
21	Kim ID, Sawicki E, Lee HK, Lee EH, Park HJ, Han PL, Kim KK, Choi H and Lee JK: Robust neuroprotective effects of intranasally delivered iNOS siRNA encapsulated in gelatin nanoparticles in the postischemic brain. Nanomedicine. 12:1219–1229. 2016. View Article : Google Scholar : PubMed/NCBI
22	Murakami K, Kondo T, Kawase M, Li Y, Sato S, Chen SF and Chan PH: Mitochondrial susceptibility to oxidative stress exacerbates cerebral infarction that follows permanent focal cerebral ischemia in mutant mice with manganese superoxide dismutase deficiency. J Neurosci. 18:205–213. 1998.PubMed/NCBI
23	Wu C, Wang P, Rao J, Wang Z, Zhang C, Lu L and Zhang F: Triptolide alleviates hepatic ischemia/reperfusion injury by attenuating oxidative stress and inhibiting NF-κB activity in mice. J Surg Res. 166:e205–e213. 2011. View Article : Google Scholar : PubMed/NCBI
24	Shiraki T, Aoyama T, Yokoyama C, Hayakawa Y, Tanaka T, Nishigaki K, Sawamura T and Minatoguchi S: LOX-1 plays an important role in ischemia-induced angiogenesis of limbs. PLoS One. 9:e1145422014. View Article : Google Scholar : PubMed/NCBI
25	Mao X, Xie L and Greenberg DA: Effects of flow on LOX-1 and oxidized low-density lipoprotein interactions in brain endothelial cell cultures. Free Radic Biol Med. 89:638–641. 2015. View Article : Google Scholar : PubMed/NCBI
26	Ozturk H, Gezici A and Ozturk H: The effect of celecoxib, a selective COX-2 inhibitor, on liver ischemia/reperfusion-induced oxidative stress in rats. Hepatol Res. 34:76–83. 2006. View Article : Google Scholar
27	Du Y, Zhu Y, Teng X, Zhang K, Teng X and Li S: Toxicological effect of manganese on NF-κB/iNOS-COX-2 signaling pathway in chicken testes. Biol Trace Elem Res. 168:227–234. 2015. View Article : Google Scholar : PubMed/NCBI
28	Weng C, Li Y, Xu D, Shi Y and Tang H: Specific cleavage of Mcl-1 by caspase-3 in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in Jurkat leukemia T cells. J Biol Chem. 280:10491–10500. 2005. View Article : Google Scholar : PubMed/NCBI
29	Lavrik IN, Golks A and Krammer PH: Caspases: Pharmacological manipulation of cell death. J Clin Invest. 115:2665–2672. 2005. View Article : Google Scholar : PubMed/NCBI
30	Stennicke HR and Salvesen GS: Biochemical characteristics of caspases-3, -6, -7, and -8. J Biol Chem. 272:25719–25723. 1997. View Article : Google Scholar : PubMed/NCBI
31	Hsu YT, Wolter KG and Youle RJ: Cytosol-to-membrane redistribution of Bax and Bcl-X(L) during apoptosis. Proc Natl Acad Sci USA. 94:3668–3672. 1997. View Article : Google Scholar : PubMed/NCBI
32	Nechushtan A, Smith CL, Hsu YT and Youle RJ: Conformation of the Bax C-terminus regulates subcellular location and cell death. EMBO J. 18:2330–2341. 1999. View Article : Google Scholar : PubMed/NCBI
33	Shi Y, Chen J, Weng C, Chen R, Zheng Y, Chen Q and Tang H: Identification of the protein-protein contact site and interaction mode of human VDAC1 with Bcl-2 family proteins. Biochem Biophys Res Commun. 305:989–996. 2003. View Article : Google Scholar : PubMed/NCBI
34	Duan M, Wang ZC, Wang XY, Shi JY, Yang LX, Ding ZB, Gao Q, Zhou J and Fan J: TREM-1, an inflammatory modulator, is expressed in hepatocellular carcinoma cells and significantly promotes tumor progression. Ann Surg Oncol. 22:3121–3129. 2015. View Article : Google Scholar