Radioprotective 105 kDa protein attenuates ischemia/reperfusion-induced myocardial apoptosis and autophagy by inhibiting the activation of the TLR4/NF-κB signaling pathway in rats

Guo,Xin; Jiang,Hong; Yang,Jun; Chen,Jing; Yang,Jian; Ding,Jia-Wang; Li,Song; Wu,Hui; Ding,Hua-Sheng

doi:10.3892/ijmm.2016.2686

Radioprotective 105 kDa protein attenuates ischemia/reperfusion-induced myocardial apoptosis and autophagy by inhibiting the activation of the TLR4/NF-κB signaling pathway in rats

Authors:
- Xin Guo
- Hong Jiang
- Jun Yang
- Jing Chen
- Jian Yang
- Jia-Wang Ding
- Song Li
- Hui Wu
- Hua-Sheng Ding
View Affiliations

Affiliations: Department of Cardiology, Renmin Hospital of Wuhan University, Cardiovascular Research Institute of Wuhan University, Hubei Key Laboratory of Cardiology, Wuhan, Hubei 430060, P.R. China, Institute of Cardiovascular Diseases, China Three Gorges University, Yichang, Hubei 443000, P.R. China
Published online on: July 19, 2016 https://doi.org/10.3892/ijmm.2016.2686
Pages: 885-893

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

Toll-like receptor 4 (TLR4) serves as an important inducer of apoptotic and autophagic responses in myocardial ischemia/reperfusion (I/R) injury (MIRI). Radioprotective 105 kDa protein (RP105) is a specific inhibitor of TLR4. However, the molecular mechanisms by which RP105 represses myocardial apoptosis and autophagy through TLR4‑mediated signaling during I/R have not yet been fully elucidated. Therefore, in the present study, we aimed to examine whether adenovirus-mediated RP105 overexpression repressed myocardial apoptosis and autophagy by inhibiting the TLR4-driven mechanism in MIRI. Three days after the injection of virus or saline into the myocardium, Sprague-Dawley (SD) rats were subjected to 30 min of left anterior descending coronary artery occlusion and 6 h of reperfusion. Myocardial specimens were prepared for analysis. We performed immunohistochemichal and histopathological analysis, the measurement of cardiac biomarkers, TUNEL assay , RT-qPCR and western blot analysis. The results indicated that the overexpression of RP105 contributed to an amelioration of myocardial histological damage, decreased leakage of creatine kinase (CK) and lactate dehydrogenase (LDH), as well as a reduction in the number of TUNEL-positive cardiomyocytes. The levels of positively associated modulators of apoptosis and autophagy were also significantly downregulated by RP105, whereas Bcl-2, which plays an opposite role in inducing apoptosis and autophagy, was inversely upregulated. Furthermore, the overexpression of RP105 led to the repression of TLR4 activity and the phosphorylation of NF-κB/p65, as well as the reduced production of the cytokines interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α). Taken together, these data suggest that RP105 protects the myocardium against apoptosis and autophagy, and plays a cardioprotective role during I/R injury. This is most likely due to the inactivation of TLR4/NF-κB signaling pathway. Thus, RP105 may represent an innovative therapeutic target for attenuating MIRI.

View Figures

View References

1	Zhou X, Chen M, Wang S, Yu L and Jiang H: MG53 protein: a promising novel therapeutic target for myocardial ischemia reperfusion injury. Int J Cardiol. 199:424–425. 2015. View Article : Google Scholar : PubMed/NCBI
2	Han XJ, He D, Xu LJ, Chen M, Wang YQ, Feng JG, Wei MJ, Hong T and Jiang LP: Knockdown of connexin 43 attenuates balloon injury-induced vascular restenosis through the inhibition of the proliferation and migration of vascular smooth muscle cells. Int J Mol Med. 36:1361–1368. 2015.PubMed/NCBI
3	Thapalia BA, Zhou Z and Lin X: Autophagy, a process within reperfusion injury: an update. Int J Clin Exp Pathol. 7:8322–8341. 2014.
4	Lin Y, Chen L, Li W and Fang J: Role of high-mobility group box-1 in myocardial ischemia/reperfusion injury and the effect of ethyl pyruvate. Exp Ther Med. 9:1537–1541. 2015.PubMed/NCBI
5	Yao T, Ying X, Zhao Y, Yuan A, He Q, Tong H, Ding S, Liu J, Peng X, Gao E, et al: Vitamin D receptor activation protects against myocardial reperfusion injury through inhibition of apoptosis and modulation of autophagy. Antioxid Redox Signal. 22:633–650. 2015. View Article : Google Scholar :
6	Barry SP, Ounzain S, McCormick J, Scarabelli TM, Chen-Scarabelli C, Saravolatz LI, Faggian G, Mazzucco A, Suzuki H, Thiemermann C, et al: Enhanced IL-17 signalling following myocardial ischaemia/reperfusion injury. Int J Cardiol. 163:326–334. 2013. View Article : Google Scholar :
7	Ding HS, Yang J, Chen P, Yang J, Bo SQ, Ding JW and Yu QQ: The HMGB1-TLR4 axis contributes to myocardial ischemia/reperfusion injury via regulation of cardiomyocyte apoptosis. Gene. 527:389–393. 2013. View Article : Google Scholar : PubMed/NCBI
8	Wang B, Zhong S, Zheng F, Zhang Y, Gao F, Chen Y, Lu B, Xu H and Shi G: N-n-butyl haloperidol iodide protects cardiomyocytes against hypoxia/reoxygenation injury by inhibiting autophagy. Oncotarget. 6:24709–24721. 2015. View Article : Google Scholar : PubMed/NCBI
9	Murphy E and Steenbergen C: Mechanisms underlying acute protection from cardiac ischemia-reperfusion injury. Physiol Rev. 88:581–609. 2008. View Article : Google Scholar : PubMed/NCBI
10	Wu H, Che X, Zheng Q, Wu A, Pan K, Shao A, Wu Q, Zhang J and Hong Y: Caspases: a molecular switch node in the crosstalk between autophagy and apoptosis. Int J Biol Sci. 10:1072–1083. 2014. View Article : Google Scholar : PubMed/NCBI
11	Kalogeris T, Baines CP, Krenz M and Korthuis RJ: Cell biology of ischemia/reperfusion injury. Int Rev Cell Mol Biol. 298:229–317. 2012. View Article : Google Scholar : PubMed/NCBI
12	Xie M, Kong Y, Tan W, May H, Battiprolu PK, Pedrozo Z, Wang ZV, Morales C, Luo X, Cho G, et al: Histone deacetylase inhibition blunts ischemia/reperfusion injury by inducing cardiomyocyte autophagy. Circulation. 129:1139–1151. 2014. View Article : Google Scholar : PubMed/NCBI
13	Gustafsson ÅB and Gottlieb RA: Eat your heart out: role of autophagy in myocardial ischemia/reperfusion. Autophagy. 4:416–421. 2008. View Article : Google Scholar : PubMed/NCBI
14	Xu W, Jiang H, Hu X and Fu W: Effects of high-mobility group box 1 on the expression of Beclin-1 and LC3 proteins following hypoxia and reoxygenation injury in rat cardiomyocytes. Int J Clin Exp Med. 7:5353–5357. 2014.
15	Ke J, Yao B, Li T, Cui S and Ding H: A2 adenosine receptor-mediated cardioprotection against reperfusion injury in rat hearts is associated with autophagy downregulation. J Cardiovasc Pharmacol. 66:25–34. 2015. View Article : Google Scholar : PubMed/NCBI
16	Nikoletopoulou V, Markaki M, Palikaras K and Tavernarakis N: Crosstalk between apoptosis, necrosis and autophagy. Biochim Biophys Acta. 1833:3448–3459. 2013. View Article : Google Scholar : PubMed/NCBI
17	Xu J, Qin X, Cai X, Yang L, Xing Y, Li J, Zhang L, Tang Y, Liu J, Zhang X and Gao F: Mitochondrial JNK activation triggers autophagy and apoptosis and aggravates myocardial injury following ischemia/reperfusion. Biochim Biophys Acta. 1852:262–270. 2015. View Article : Google Scholar
18	Shen M, Lu J, Dai W, Wang F, Xu L, Chen K, He L, Cheng P, Zhang Y, Wang C, et al: Ethyl pyruvate ameliorates hepatic ischemia-reperfusion injury by inhibiting intrinsic pathway of apoptosis and autophagy. Mediators Inflamm. 2013:4615362013. View Article : Google Scholar
19	Zhao Y, Xu Y, Zhang J and Ji T: Cardioprotective effect of carvedilol: inhibition of apoptosis in H9c2 cardiomyocytes via the TLR4/NF-κB pathway following ischemia/reperfusion injury. Exp Ther Med. 8:1092–1096. 2014.PubMed/NCBI
20	Xu Y, Jagannath C, Liu XD, Sharafkhaneh A, Kolodziejska KE and Eissa NT: Toll-like receptor 4 is a sensor for autophagy associated with innate immunity. Immunity. 27:135–144. 2007. View Article : Google Scholar : PubMed/NCBI
21	Porakishvili N, Vispute K, Steele AJ, Rajakaruna N, Kulikova N, Tsertsvadze T, Nathwani A, Damle RN, Clark EA, Rai KR, et al: Rewiring of sIgM-mediated intracellular signaling through the CD180 Toll-like receptor. Mol Med. 21:46–57. 2015. View Article : Google Scholar :
22	Wezel A, van der Velden D, Maassen JM, Lagraauw HM, de Vries MR, Karper JC, Kuiper J, Bot I and Quax PH: RP105 deficiency attenuates early atherosclerosis via decreased monocyte influx in a CCR2 dependent manner. Atherosclerosis. 238:132–139. 2015. View Article : Google Scholar
23	Louwe MC, Karper JC, de Vries MR, Nossent AY, Bastiaansen AJ, van der Hoorn JW, Willems van Dijk K, Rensen PC, Steendijk P, Smit JW and Quax PH: RP105 deficiency aggravates cardiac dysfunction after myocardial infarction in mice. Int J Cardiol. 176:788–793. 2014. View Article : Google Scholar : PubMed/NCBI
24	Karper JC, Ewing MM, de Vries MR, de Jager SC, Peters EA, de Boer HC, van Zonneveld AJ, Kuiper J, Huizinga EG, Brondijk TH, et al: TLR accessory molecule RP105 (CD180) is involved in post-interventional vascular remodeling and soluble RP105 modulates neointima formation. PLoS One. 8:e679232013. View Article : Google Scholar : PubMed/NCBI
25	Yang J, Guo X, Yang J, Ding JW, Li S, Yang R, Fan ZX and Yang CJ: RP105 protects against apoptosis in ischemia/reperfusion-induced myocardial damage in rats by suppressing TLR4-mediated signaling pathways. Cell Physiol Biochem. 36:2137–2148. 2015. View Article : Google Scholar : PubMed/NCBI
26	Yang J, Jiang H, Chen SS, Chen J, Li WQ, Xu SK and Wang JC: Lentivirus-mediated RNAi targeting CREB binding protein attenuates neointimal formation and promotes re-endothelialization in balloon injured rat carotid artery. Cell Physiol Biochem. 26:441–448. 2010. View Article : Google Scholar
27	Zeng M, Wei X, Wu Z, Li W, Li B, Zhen Y, Chen J, Wang P and Fei Y: NF-κB-mediated induction of autophagy in cardiac ischemia/reperfusion injury. Biochem Biophys Res Commun. 436:180–185. 2013. View Article : Google Scholar : PubMed/NCBI
28	Shyu KG, Wang BW, Lin CM and Chang H: Cyclic stretch enhances the expression of toll-like receptor 4 gene in cultured cardiomyocytes via p38 MAP kinase and NF-kappaB pathway. J Biomed Sci. 17:152010. View Article : Google Scholar : PubMed/NCBI
29	Eltzschig HK and Eckle T: Ischemia and reperfusion - from mechanism to translation. Nat Med. 17:1391–1401. 2011. View Article : Google Scholar : PubMed/NCBI
30	Divanovic S, Trompette A, Atabani SF, Madan R, Golenbock DT, Visintin A, Finberg RW, Tarakhovsky A, Vogel SN, Belkaid Y, et al: Negative regulation of Toll-like receptor 4 signaling by the Toll-like receptor homolog RP105. Nat Immunol. 6:571–578. 2005. View Article : Google Scholar : PubMed/NCBI
31	Yazawa N, Fujimoto M, Sato S, Miyake K, Asano N, Nagai Y, Takeuchi O, Takeda K, Okochi H, Akira S, et al: CD19 regulates innate immunity by the toll-like receptor RP105 signaling in B lymphocytes. Blood. 102:1374–1380. 2003. View Article : Google Scholar : PubMed/NCBI
32	Hsieh YC, Athar M and Chaudry IH: When apoptosis meets autophagy: deciding cell fate after trauma and sepsis. Trends Mol Med. 15:129–138. 2009. View Article : Google Scholar : PubMed/NCBI
33	Yang Y, Gao K, Hu Z, Li W, Davies H, Ling S, Rudd JA and Fang M: Autophagy upregulation and apoptosis downregulation in DAHP and triptolide treated cerebral ischemia. Mediators Inflamm. 2015:1201982015. View Article : Google Scholar : PubMed/NCBI
34	Maejima Y, Kyoi S, Zhai P, Liu T, Li H, Ivessa A, Sciarretta S, Del Re DP, Zablocki DK, Hsu CP, et al: Mst1 inhibits autophagy by promoting Beclin1-Bcl-2 interaction. Nat Med. 19:1478–1488. 2013. View Article : Google Scholar : PubMed/NCBI