RP105 ameliorates hypoxia̸reoxygenation injury in cardiac microvascular endothelial cells by suppressing TLR4̸MAPKs̸NF-κB signaling

  • Authors:
    • Xin Guo
    • Hong Jiang
    • Jing Chen
    • Bo-Fang Zhang
    • Qi Hu
    • Shuo Yang
    • Jian Yang
    • Jing Zhang
  • View Affiliations

  • Published online on: April 16, 2018     https://doi.org/10.3892/ijmm.2018.3621
  • Pages: 505-513
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Abstract

The radioprotective 105 kDa protein (RP105) has been implicated in the pathological process of multiple cardiovascular diseases through its functional and physical interactions with Toll‑like receptor 4 (TLR4). However, the effects of RP105 on cardiac microvascular endothelial cells (CMECs) in response to hypoxia̸reoxygenation (H̸R) injury have not been extensively investigated. The aim of the present study was to elucidate the potential roles of RP105 in the protection of CMECs against H̸R injury, and investigate the underlying mechanisms. CMECs isolated from Sprague‑Dawley rats were transduced with adenoviral vectors encoding RP105 or green fluorescent protein (GFP). At 48 h post‑transfection, CMECs were subjected to hypoxia for 4 h and reoxygenation for 2 h (H̸R) to simulate the in vivo ischemia̸reperfusion model. The mRNA and protein levels of RP105 were detected by reverse transcription‑quantitative polymerase chain reaction and western blot analysis, respectively. The effects of RP105 on CMEC proliferation, migration and apoptosis were measured by GFP‑8, Transwell chamber and flow cytometry assays, respectively. The secretion of interleukin (IL)‑6 and tumor necrosis factor (TNF)‑α in the culture medium was measured by ELISA. Moreover, the expression level of TLR4, p38 mitogen‑activated protein kinase (MAPK), extracellular-signal-regulated kinase 1̸2, c-Jun N-terminal kinase, nuclear factor (NF)‑κB̸p65, IL‑6, TNF‑α and intercellular adhesion melecule‑1 was evaluated by western blot analysis. The results demonstrated that RP105 was minimally expressed in CMECs subjected to H̸R injury. Overexpression of RP105 via adenoviral vectors was able to significantly protect CMECs against H̸R injury, as evidenced by the promotion of cell proliferation and migration, as well as the amelioration of inflammation and apoptosis. These beneficial effects were at least partly mediated through inhibition of TLR4̸MAPKs̸NF‑κB signaling. Therefore, RP105 may be a promising candidate for prevention against CMECs‑associated H̸R injury.

References

1 

Ibanez B, Heusch G, Ovize M and Van de Werf F: Evolving therapies for myocardial ischemia̸reperfusion injury. J Am Coll Cardiol. 65:1454–1471. 2015. View Article : Google Scholar

2 

Zhang Y, Zhou H, Wu W, Shi C, Hu S, Yin T, Ma Q, Han T, Zhang Y, Tian F, et al: Liraglutide protects cardiac microvascular endothelial cells against hypoxia/reoxygenation injury through the suppression of the SR-Ca(2+)-XO-ROS axis via activation of the GLP-1R̸PI3K̸Akt̸survivin pathways. Free Radic Biol Med. 95:278–292. 2016. View Article : Google Scholar : PubMed/NCBI

3 

Cui H, Li X, Li N, Qi K, Li Q, Jin C, Zhang Q, Jiang L and Yang Y: Induction of autophagy by Tongxinluo through the MEK̸ERK pathway protects human cardiac microvascular endothelial cells from hypoxia/reoxygenation injury. J Cardiovasc Pharmacol. 64:180–190. 2014. View Article : Google Scholar : PubMed/NCBI

4 

Liu Y, Lian K, Zhang L, Wang R, Yi F, Gao C, Xin C, Zhu D, Li Y, Yan W, et al: TXNIP mediates NLRP3 inflammasome activation in cardiac microvascular endothelial cells as a novel mechanism in myocardial ischemia/reperfusion injury. Basic Res Cardiol. 109:4152014. View Article : Google Scholar : PubMed/NCBI

5 

Li JM, Mullen AM and Shah AM: Phenotypic properties and characteristics of superoxide production by mouse coronary microvascular endothelial cells. J Mol Cell Cardiol. 33:1119–1131. 2001. View Article : Google Scholar : PubMed/NCBI

6 

Zhou Y, Zhang Y, Gao F, Guo F, Wang J, Cai W, Chen Y, Zheng J and Shi G: N-n-butyl haloperidol iodide protects cardiac microvascular endothelial cells from hypoxia/reoxygenation injury by downregulating Egr-1 expression. Cell Physiol Biochem. 26:839–848. 2010. View Article : Google Scholar

7 

Qi XF, Li YJ, Chen ZY, Kim SK, Lee KJ and Cai DQ: Involvement of the FoxO3a pathway in the ischemia/reperfusion injury of cardiac microvascular endothelial cells. Exp Mol Pathol. 95:242–247. 2013. View Article : Google Scholar : PubMed/NCBI

8 

Wang J, Hong Z, Zeng C, Yu Q and Wang H: NADPH oxidase 4 promotes cardiac microvascular angiogenesis after hypoxia/reoxygenation in vitro. Free Radic Biol Med. 69:278–288. 2014. View Article : Google Scholar : PubMed/NCBI

9 

Coulombe KL, Bajpai VK, Andreadis ST and Murry CE: Heart regeneration with engineered myocardial tissue. Annu Rev Biomed Eng. 16:1–28. 2014. View Article : Google Scholar : PubMed/NCBI

10 

Zhang Z, Li W, Sun D, Zhao L, Zhang R, Wang Y, Zhou X, Wang H and Cao F: Toll-like receptor 4 signaling in dysfunction of cardiac microvascular endothelial cells under hypoxia/reoxygenation. Inflamm Res. 60:37–45. 2011. View Article : Google Scholar

11 

Guo X, Jiang H and Chen J: RP105-PI3K-Akt axis: A potential therapeutic approach for ameliorating myocardial ischemia̸reperfusion injury. Int J Cardiol. 206:95–96. 2016. View Article : Google Scholar : PubMed/NCBI

12 

Hinkel R, Lange P, Petersen B, Gottlieb E, Ng JK, Finger S, Horstkotte J, Lee S, Thormann M, Knorr M, et al: Heme oxygenase-1 gene therapy provides cardioprotection via control of post-ischemic inflammation: An experimental study in a pre- clinical pig model. J Am Coll Cardiol. 66:154–165. 2015. View Article : Google Scholar : PubMed/NCBI

13 

Zanotti G, Casiraghi M, Abano JB, Tatreau JR, Sevala M, Berlin H, Smyth S, Funkhouser WK, Burridge K, Randell SH, et al: Novel critical role of Toll-like receptor 4 in lung ischemia-reperfusion injury and edema. Am J Physiol Lung Cell Mol Physiol. 297:L52–L63. 2009. View Article : Google Scholar : PubMed/NCBI

14 

Chen Y, Huang XJ, Yu N, Xie Y, Zhang K, Wen F, Liu H and Di Q: HMGB1 contributes to the expression of P-Glycoprotein in mouse epileptic brain through Toll-Like receptor 4 and receptor for advanced glycation end products. PLoS One. 10:e01409182015. View Article : Google Scholar : PubMed/NCBI

15 

Mountain DJ, Singh M and Singh K: Downregulation of VEGF-D expression by interleukin-1beta in cardiac microvascular endothelial cells is mediated by MAPKs and PKCalpha̸beta1. J Cell Physiol. 215:337–343. 2008. View Article : Google Scholar

16 

Liu RR, Li J, Gong JY, Kuang F, Liu JY, Zhang YS, Ma QL, Song CJ, Truax AD, Gao F, et al: MicroRNA-141 regulates the expression level of ICAM-1 on endothelium to decrease myocardial ischemia-reperfusion injury. Am J Physiol Heart Circ Physiol. 309:H1303–H1313. 2015. View Article : Google Scholar : PubMed/NCBI

17 

Liu B, Zhang N, Liu Z, Fu Y, Feng S, Wang S, Cao Y, Li D, Liang D, Li F, et al: RP105 involved in activation of mouse macrophages via TLR2 and TLR4 signaling. Mol Cell Biochem. 378:183–193. 2013. View Article : Google Scholar : PubMed/NCBI

18 

Li X, Yang J, Yang J, Dong W, Li S, Wu H and Li L: RP105 protects against myocardial ischemia-reperfusion injury via suppressing TLR4 signaling pathways in rat model. Exp Mol Pathol. 100:281–286. 2016. View Article : Google Scholar : PubMed/NCBI

19 

Hijiya N, Miyake K, Akashi S, Matsuura K, Higuchi Y and Yamamoto S: Possible involvement of toll-like receptor 4 in endothelial cell activation of larger vessels in response to lipo-polysaccharide. Pathobiology. 70:18–25. 2002. View Article : Google Scholar

20 

Wezel A, de Vries MR, Maassen JM, Kip P, Peters EA, Karper JC, Kuiper J, Bot I and Quax PH: Deficiency of the TLR4 analogue RP105 aggravates vein graft disease by inducing a pro-inflammatory response. Sci Rep. 6:242482016. View Article : Google Scholar : PubMed/NCBI

21 

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

22 

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, et al: RP105 deficiency aggravates cardiac dysfunction after myocardial infarction in mice. Int J Cardiol. 176:788–793. 2014. View Article : Google Scholar : PubMed/NCBI

23 

Xia JB, Liu GH, Chen ZY, Mao CZ, Zhou DC, Wu HY, Park KS, Zhao H, Kim SK, Cai DQ, et al: Hypoxia̸ischemia promotes CXCL10 expression in cardiac microvascular endothelial cells by NFkB activation. Cytokine. 81:63–70. 2016. View Article : Google Scholar : PubMed/NCBI

24 

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

25 

Yang J, Chen L, Ding J, Zhang J, Fan Z, Yang C, Yu Q and Yang J: Cardioprotective effect of miRNA-22 on hypoxia/reoxygenation induced cardiomyocyte injury in neonatal rats. Gene. 579:17–22. 2016. View Article : Google Scholar

26 

Choi HS, Kim MK, Choi YK, Shin YC, Cho SG and Ko SG: Rhus verniciflua Stokes (RVS) and butein induce apoptosis of paclitaxel-resistant SKOV-3̸PAX ovarian cancer cells through inhibition of AKT phosphorylation. BMC Complement Altern Med. 16:1222016. View Article : Google Scholar

27 

Ye EA and Steinle JJ: miR-146a attenuates inflammatory pathways mediated by TLR4̸NF-κB and TNFα to protect primary human retinal microvascular endothelial cells grown in high glucose. Mediators Inflamm. 2016:39584532016. View Article : Google Scholar

28 

Shi J, Zhou J and Zhang M: Microcystins induces vascular inflammation in human umbilical vein endothelial cells via activation of NF-κB. Mediators Inflamm. 942159:20152015.

29 

Mudaliar H, Pollock C, Ma J, Wu H, Chadban S and Panchapakesan U: The role of TLR2 and 4-mediated inflammatory pathways in endothelial cells exposed to high glucose. PLoS One. 9:e1088442014. View Article : Google Scholar : PubMed/NCBI

30 

Sawa Y, Ueki T, Hata M, Iwasawa K, Tsuruga E, Kojima H, Ishikawa H and Yoshida S: LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 expression in human lymphatic endothelium. J Histochem Cytochem. 56:97–109. 2008. View Article : Google Scholar

31 

Li F, Li W, Li X, Li F, Zhang L, Wang B, Huang G, Guo X, Wan L, Liu Y, et al: Geniposide attenuates inflammatory response by suppressing P2Y14 receptor and downstream ERK1/2 signaling pathway in oxygen and glucose deprivation-induced brain microvascular endothelial cells. J Ethnopharmacol. 185:77–86. 2016. View Article : Google Scholar : PubMed/NCBI

32 

Cheng F, Lan J, Xia W, Tu C, Chen B, Li S and Pan W: Folic acid attenuates vascular endothelial cell injury caused by hypoxia via the inhibition of ERK1̸2̸NOX4̸ROS pathway. Cell Biochem Biophys. 74:205–211. 2016. View Article : Google Scholar : PubMed/NCBI

33 

Li J, Zhang Z, Lv L, Qiao H, Chen X and Zou C: (−)-Epigallocatechin gallate inhibits asymmetric dimethylarginine-induced injury in human brain microvascular endothelial cells. Neurochem Res. 41:1868–1876. 2016. View Article : Google Scholar : PubMed/NCBI

34 

Wang LW, Chang YC, Chen SJ, Tseng CH, Tu YF, Liao NS, Huang CC and Ho CJ: TNFR1-JNK signaling is the shared pathway of neuroinflammation and neurovascular damage after LPS-sensitized hypoxic-ischemic injury in the immature brain. J Neuroinflammation. 11:2152014. View Article : Google Scholar : PubMed/NCBI

35 

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

36 

Nagai Y, Watanabe Y and Takatsu K: The TLR family protein RP105̸MD-1 complex: A new player in obesity and adipose tissue inflammation. Adipocyte. 2:61–66. 2013. View Article : Google Scholar : PubMed/NCBI

37 

Jin C, Cleveland JC, Ao L, Li J, Zeng Q, Fullerton DA and Meng X: Human myocardium releases heat shock protein 27 (HSP27) after global ischemia: The proinflammatory effect of extracellular HSP27 through toll-like receptor (TLR)-2 and TLR4. Mol Med. 20:280–289. 2014. View Article : Google Scholar : PubMed/NCBI

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Copy and paste a formatted citation
APA
Guo, X., Jiang, H., Chen, J., Zhang, B., Hu, Q., Yang, S. ... Zhang, J. (2018). RP105 ameliorates hypoxia̸reoxygenation injury in cardiac microvascular endothelial cells by suppressing TLR4̸MAPKs̸NF-κB signaling. International Journal of Molecular Medicine, 42, 505-513. https://doi.org/10.3892/ijmm.2018.3621
MLA
Guo, X., Jiang, H., Chen, J., Zhang, B., Hu, Q., Yang, S., Yang, J., Zhang, J."RP105 ameliorates hypoxia̸reoxygenation injury in cardiac microvascular endothelial cells by suppressing TLR4̸MAPKs̸NF-κB signaling". International Journal of Molecular Medicine 42.1 (2018): 505-513.
Chicago
Guo, X., Jiang, H., Chen, J., Zhang, B., Hu, Q., Yang, S., Yang, J., Zhang, J."RP105 ameliorates hypoxia̸reoxygenation injury in cardiac microvascular endothelial cells by suppressing TLR4̸MAPKs̸NF-κB signaling". International Journal of Molecular Medicine 42, no. 1 (2018): 505-513. https://doi.org/10.3892/ijmm.2018.3621