Open Access

Protective effect of rivaroxaban on arteriosclerosis obliterans in rats through modulation of the toll‑like receptor 4/NF‑κB signaling pathway

  • Authors:
    • Xinjiang Lou
    • Zhi Yu
    • Xiaoxia Yang
    • Jie Chen
  • View Affiliations

  • Published online on: July 3, 2019     https://doi.org/10.3892/etm.2019.7726
  • Pages: 1619-1626
  • Copyright: © Lou 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: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The aim of the present study was to explore the pharmacological role of rivaroxaban in rats with arteriosclerosis obliterans (ASO) and the potential mechanism of its action. A total of 60 adult male Sprague Dawley (weighing 210‑250 g) were randomly assigned into either the sham group, model group or Riv group. Rats in the sham group were fed a normal diet, whereas those in model group and Riv group were fed a high‑fat diet for 8 weeks. After establishment of the ASO model, rats in the Riv group were intragastrically administered 10 mg/kg rivaroxaban, whereas those in the sham group and the model group were administrated with the same volume of 0.9% saline for 4 weeks. At the end of animal procedures, a blood sample and the femoral artery of the rats were harvested. The results of the present study revealed that rats in the model group presented with an irregularly narrowed femoral artery lumen, disordered endothelial cells, internal elastic plates and smooth muscle cells. By comparison, the arterial wall structure and stenosis of the femoral artery of rats in Riv group recovered and all the pathological changes were alleviated after rivaroxaban treatment. Levels of total cholesterol, triglycerides and low‑density lipoproteins decreased, whereas the level of high‑density lipoproteins increased in the Riv group compared with the model group. Rivaroxaban treatment significantly reduced serum levels of interleukin‑1, tumor necrosis factor‑α and monocyte chemoattractant protein‑1 (MCP‑1), and increased the serum level of transforming growth factor‑β (TGF‑β). Rats in the Riv group had reduced expression of toll‑like receptor 4 (TLR4), NF‑κB and MCP‑1, and increased expression of TGF‑β in femoral artery tissues compared with the model group. Therefore rivaroxaban may have exerted its anti‑atherosclerotic effects by regulating the expression of genes in the TLR4/NF‑κB signaling pathway and the activation of the downstream molecules.

Introduction

In arteriosclerosis obliterans (ASO), lipids are persistently deposited in the intima of arteries to form atheromatous plaques (1). The intima and middle layers of the arteries are deteriorated and then proliferate, leading to thickening, stiffness and distortion of arterial walls (1,2). The gradual loss of elasticity, enlargement of atherosclerotic plaque and secondary thrombosis result in narrowing or even obstruction of the arterial lumen, leading to corresponding ischemic symptoms at the distal end of arteries (1,2).

There are numerous hypotheses regarding the etiology of ASO, including lipid infiltration, thrombosis and inflammatory injury response (35). Although these hypotheses do not comprehensively explain all the pathological phenomena of ASO, they do demonstrate that atherosclerosis (AS) is initiated by form damaging stimuli, such as dysregulation of lipid metabolism, hemodynamic damage, heredity, infection, and physical or chemical stimuli (6,7). Multiple inflammatory factors and associated cytokine networks co-operatively act on the vascular wall, leading to a persistent state of vascular dysfunction (8). The gradual formation and development of AS plaques in blood vessels, accompanied by the rupture of unstable plaques and thrombosis ultimately result in different degrees of stenosis or occlusion of the arteries, leading to clinical events of acute and chronic limb ischemia (8).

Recent studies have suggested that toll-like receptor 4 (TLR4) and its associated signal transduction pathways are critical for formation of AS (9,10). TLR4 expression is high in the atherosclerotic plaque, resulting in the synthesis and release of various cytokines or chemokines associated with AS (11,12). TLR4 activates nuclear translocation of NF-κB by mediating the myeloid differentiation primary response protein MyD88 (Myd88)-dependent early response pathway, thus initiating a series of inflammatory responses by producing pro-inflammatory factors and monocyte chemoattractants (12). Transforming growth factor-β (TGF-β) is inhibited by the p38 mitogen activated protein kinase (MAPK) pathway (1315). Increased expression of monocyte chemoattractant protein-1 (MCP-1) accelerates the progression of AS, and its absence can slow the progression of AS (16). TGF-β downregulated the levels of cytokines during the atherosclerotic inflammatory response, including tumor-necrosis factor-α (TNF-α), interferon (IFN)γ and interleukin (IL)-1 (17). IL-1β and TNF-α are proinflammatory cytokines implicated in the pathogenesis of autoimmune diseases such as rheumatoid arthritis, whereas TGF-β is an anti-inflammatory cytokine, which has been reported to serve an anti-inflammatory role in autoimmune diseases such as multiple sclerosis and mediate the beneficial effect of IFNβ in multiple sclerosis (1820).

At present, the primary treatment options used for treating ASO are drug therapy and surgical treatment for inhibiting arterial intimal hyperplasia (1). However, surgical treatment has certain risks such as angina pectoris, myocardial infarction and pulmonary infection, is expensive, and the middle-aged and elderly patients may refuse surgery (21). Therefore patients with ASO are frequently treated with drug therapy including antiplatelet drugs, vasodilators and drugs that promote collateral circulation. Rivaroxaban is a novel anticoagulant with the advantages of easy absorption, a quick onset of effect and fewer and less egregious side effects (22). Rivaroxaban is used at present to prevent the formation of venous thrombosis and pulmonary embolism after hip or knee joint replacement and may also be used to prevent the recurrence of coronary artery syndrome (22). Rivaroxaban may reduce NF-κB activity through the Myd88-dependent pathway of TLR4, thereby preventing AS by reducing the expressions and release of downstream pro-inflammatory cytokines (23). The present study developed a rat model of ASO and the pharmacological role of rivaroxaban was determined when used to treat ASO.

Materials and methods

In vivo ASO model

A total of 60 adult male Sprague Dawley rats (age, 6–8 weeks; weight, 210–250 g), were obtained from Charles River Laboratories. The rats were housed in a temperature-controlled room (21±2°C) with 40–70% relative humidity and a 12-h light/dark cycle. All rats had free access to water and food. The rats were randomly assigned into three groups, a sham group, model group and the Riv group, with 20 rats in each group. Rats in the sham group were fed with a normal diet, whereas those in the model group and Riv group were fed a high-fat diet for 8 weeks. After establishment of the ASO model by surgery, rats in the Riv group were intragastrically administered with 10 mg/kg rivaroxaban as described previously (24), whereas those in sham group and model group were administrated with the same volume of 0.9% saline for 4 weeks. The present study was approved by The Animal Ethics Committee of Zhejiang Chinese Medical University Animal Center (Hangzhou, China).

Construction of ASO model in rats

After 8 weeks of high fat diet and prior to rav administration, rats were anesthetized with intraperitoneal injection of 10% chloral hydrate (300 mg/kg). After routine disinfection of the bilateral inguinal region and the skin on the inner side of the hind limbs, a longitudinal incision of ~2 cm in length was made from the bilateral groin to the knee joint. The surrounding tissue of the vascular nerve sheath was isolated, and the femoral artery and its branches were ligated for 30 sec. Rats in the sham group were only cut open but the ligation was not performed. After the incision was closed, 2 ml saline was subcutaneously injected for fluid infusion. Rats were returned to the cage until their vital signs were stable.

Sample collection

After a total of 4 weeks of Rav/saline treatment, the rats received anesthesia with 10% chloral hydrate (0.4 g/kg) and were sacrificed by cervical dislocation prior to blood collection. A 2.5-ml blood sample was harvested from the tail vein and centrifuged at 1,000 × g for 15 min at 4°C The supernatant was collected and preserved at −80°C. A part of rat femoral artery was resected and washed with PBS. Half of the femoral artery was preserved at −80°C, and the other half was fixed in 4% paraformaldehyde or 2.5% glutaraldehyde at room temperature for 24 h.

Hematoxylin and eosin (H&E) staining

The rat femoral artery fixed with 4% paraformaldehyde was embedded in paraffin and sectioned into slices (4-µm-thick). Artery slices were unfolded in warm water at room temperature for 60 min and transferred on to a slide. After staining with hematoxylin for 5 min at room temperature and eosin for 3 min at room temperature, the femoral artery was observed using a light microscope (magnification, ×400).

Transmission electron microscopy

Rat femoral arteries fixed with 2.5% glutaraldehyde were washed with PBS and re-fixed with 1% osmic acid at room temperature for 2 h. After washing with PBS and different concentrations of ethanol (30–100%), the femoral artery was dehydrated using 100% acetone and embedded in Epon 812 at 45°C for 12 h. The artery was sectioned in to 70-nm thick slices. Sections were stained with lead citrate for 10 min and uranium acetate for 30 min at room temperature, and finally imaged using transmission electron microscope (magnification, ×25,000).

Determination of the serum lipid levels

Serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) in rats were measured using an automatic serum biochemical analyzer (BK-200; BIOBASE).

ELISA

Serum samples were warmed in room temperature. Serum levels of IL-1 (cat. no. SRLB00), TNF-α (cat. no. SRTA00), MCP-1 (cat. no. DY3144-05) and TGF-β (cat. no. SMB100B) in rats were determined using commercial ELISA kits (R&D Systems, Inc.).

Western blotting

The femoral artery tissues were homogenized after the addition of RIPA lysis buffer (Beyotime Institute of Biotechnology). Then the supernatant was centrifuged at 5,000 × g for 10 min at 4°C. BCA assay kit (cat. no. p0011; Beyotime Institute of Biotechnology) was used to determine total protein content. A total of 10 µg protein was loaded per lane and separated by SDS-PAGE (12% gel). After being separated, the proteins were transferred to a PVDF membrane (Roche Diagnostics), which was blocked with 5% skim milk for 1 h at room temperature. The specific primary antibodies including TLR4 (1:500; cat. no. ab217274; Abcam), NF-κB (1:500; cat. no. ab32360; Abcam), MCP-1 (1:500; cat. no. ab25124; Abcam), TGF-β (1:500; cat. no. ab92486; Abcam) and GAPDH (1:500; cat. no. ab181602; Abcam), were used to incubate with the membrane overnight at 4°C. The membrane was washed with TBS with Tween-20 (TBST) five times, and the goat anti-rabbit IgG H&L secondary antibody (1:1,000; cat. no. ab150077; Abcam) was used to incubate the membrane for 2 h at room temperature. After washing with TBST for 1 min, SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific, Inc.) was used to visualize the signals. Quantity One software (version 4.0, Bio-Rad Laboratories, Inc.) was used for quantification.

Statistical analyses

All statistical analyses were performed in SPSS version 17.0 (SPSS Inc.). Data are presented as the mean ± standard deviation. Differences among multiple groups were analyzed by one-way ANOVA, followed by a least significant difference post-hoc test. Each experiment was repeated three times. P<0.05 was considered to indicate a statistically significant difference.

Results

Pathological changes of rat femoral artery

Femoral artery endothelial cells, inner elastic plate and smooth muscle cells of rats in the sham group were regularly arranged, and the vascular lumen was not narrowed (Fig. 1A). Rats in model group presented irregularly narrowed femoral artery lumen (black arrow), disordered endothelial cells (red arrow), defective internal elastic lamina (blue arrow) and proliferative smooth muscle cells (green arrow) (Fig. 1B). These characteristics were present in the Riv group, but to a lesser extent compared with the model group indicating that the arterial wall structure and stenosis of the femoral artery of rats in Riv group were partially recovered (Fig. 1C).

Transmission electron microscopy of the rat femoral artery

Endothelial cells of rats in the sham group were intact and covered the vascular surface. The internal elastic lamina was clear and complete (Fig. 2A). In the model group, the endothelial cells were irregular and the incomplete endothelium was exposed to the luminal surface (blue arrow). The inner elastic plate was partially deformed (red arrow). Smooth muscle cells, which had migrated to the vascular intima, showed deformation and nuclear condensation (green arrow) (Fig. 2B). Rats in Riv group presented regular arterial endothelial cells and smooth muscle cells, as well as a continuous elastic plate (Fig. 2C).

Serum lipid levels

Compared with the sham group, rats in the model group exhibited significantly higher levels of TC, TG and LDL-C, as well as significantly lower levels of HDL-C (all P<0.05; Fig. 3). The levels of TC, TG and LDL-C were significantly decreased, and the levels of HDL-C were significantly increased in the Riv group compared with the model group (all P<0.05; Fig. 3).

Serum levels of inflammatory factors in rats of different groups

ELISA data demonstrated that rats in the model group exhibited significantly higher serum levels of IL-1, TNF-α and MCP-1 compared with the sham group, whereas the TGF-β level was significantly lower (all P<0.05; Fig. 4). Rivaroxaban treatment significantly decreased the serum levels of IL-1, TNF-α and MCP-1, and increased the serum levels of TGF-β compared with the model group (all P<0.05; Fig. 4).

Protein expression of TLR4, NF-κB, MCP-1 and TGF-β in rat femoral artery

Upregulated protein expression levels of TLR4, NF-κB and MCP-1 were detected by western blot analysis the in femoral artery tissues of rats in the model group compared with those in sham group (all P<0.05; Fig. 5). However, rats in the Riv group exhibited decreased expression of TLR4, NF-κB and MCP-1 in the femoral artery tissues compared with the model group (all P<0.05; Fig. 5). Protein expression levels of TGF-β were lower in the femoral artery tissues of rats in the model group compared with the sham group, and were increased in the Riv group compared to the model group (all P<0.05, Fig. 5).

Discussion

The occurrence of ASO is a complex process, and its cause has not been fully elucidated. Genetics, sex, age, abnormal lipid metabolism, obesity, smoking, mechanical damage of blood vessel walls and imbalance of trace elements are recognized as factors affecting the occurrence of ASO (25). Additionally, long-term mental stimulation and emotional stress result in contraction of the arteries (26). Increased blood pressure results in dystrophy of the blood vessel wall and deposition of certain substances in the blood vessels, eventually leading to the occurrence of ASO (2527).

The inflammatory response is an important factor affecting the occurrence of ASO. TLRs are not only key molecules of the inflammatory process, but are also the initial link between the recognition of exogenous antigens and initiation of the inflammatory response (28). TLRs are a family of receptors expressed on the cell membrane. As transmembrane signal transduction receptors, TLRs link both innate and specific immunity, when molecular components of certain microorganisms are recognized (29). In vitro studies have shown that TLR4 expression is lower in human vascular endothelial cells under physiological conditions (14,15). However, stimulation of inflammatory factors markedly upregulates TLR-4 expression in tunica media vascular smooth muscle cells, exerting a significant role in vascular reconstruction (30). TLR4 is expression is increased in human atherosclerotic plaques, and is involved in the proliferative regulation of smooth muscle cells (28). Plaques subsequently migrate to the tunica intima under stimulation of cytokines, which is the primary step in the formation of an atherosclerotic plaque (31,32). NF-κB is an essential multi-channel nuclear transcription factor involved in the inflammatory process, cell proliferation and differentiation (33,34). TLR4 not only activates NF-κB, but also stimulates macrophage aggregation and inflammatory response by upregulating MCP-1 through the Myd88-dependent signaling pathway (16,35). MCP-1 is involved in the formation and transformation of macrophages and may promote the formation of atherosclerotic plaques by regulating inflammatory factors (35).

Rivaroxaban is a highly selective oral drug that directly inhibits factor Xa (FXa), which has an antithrombotic effect in an in vivo arteriovenous thrombosis model. Rivaroxaban not only inhibits free FXa, but also inhibits the activity of FXa in the prothrombin complex (36). In the coagulation cascade, FXa is involved in regulating the conversion of prothrombin to thrombin on the surface of vascular cells (36). A previous study demonstrated that FXa activates the acute inflammatory response (37). In endothelial cells, FXa can activate NF-κB, resulting in the release of inflammatory factors such as IL-6 and MCP-1 (38). Activation of inflammatory pathways is closely associated with the coagulation reaction (37,38). Previous studies have found that anticoagulant therapy efficiently inhibits coagulation activation and the inflammatory response, suggesting that anticoagulant therapy may be applied in treating ASO (39).

In the present study, the levels of IL-1, TNF-α, MCP-1, TLR-4 and NF-κB in the rats of the model group were significantly increased compared with those in the sham group, whereas TGF-β levels decreased. TGF-β expression may be inhibited by Myd88-dependent TLR4/NF-κB signal transduction by activating the p38MAPK pathway, thus attenuating the anti-inflammatory effect of TGF-β (9). Levels of IL-1, TNF-α, MCP-1, TLR-4 and NF-κB in the Riv group were lower compared with those in the model group, while the TGF-β level increased. Therefore, rivaroxaban may suppress transcriptional activity of NF-κB and synthesis of MCP-1 by inhibiting TLR4 expression. TGF-β expression was increased in the Riv group, which in turn negatively regulated Myd88-dependent TLR4/NF-κB signal transduction decreasing the inflammatory response. Specific TLR4 inhibitors, such as VGX-1027 and eritoranor, were used to treat a number of inflammatory diseases, with positive results (4042). The results of the present study highlight the possibility of using specific TLR4 inhibitors to treat ASO.

In conclusion, an ASO model in rats was developed by crush injury of the femoral artery and feeding the rats with a high-fat diet. The TLR4/NF-κB pathway and its downstream inflammatory factors were inhibited following rivaroxaban treatment. Therefore, rivaroxaban may prevent ASO through inhibiting inflammatory response.

The TLR-4/NF-κB signaling pathway is an important signal transduction mechanism and may be a key regulatory pathway in AOS. Rivaroxaban may significantly inhibit inflammation and serve an anti-atherosclerotic role by inhibiting the TLR-4/NF-κB signaling pathway and downstream inflammatory factors.

Acknowledgements

Not applicable.

Funding

No funding was received.

Availability of data and materials

The datasets used and/or analyzed during the present study are available from the corresponding author on reasonable request.

Authors' contributions

XL and XY designed the study, performed the experiments, analyzed the data, and prepared the manuscript. XL, JC and ZY established the animal models and collected the data. All authors read and approved the final manuscript.

Ethics approval and consent to participate

The present study was approved by the Animal Ethics Committee of Zhejiang Chinese Medical University Animal Center (Hangzhou, China; approval no. 20180322).

Patient consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

References

1 

Akagi D, Hoshina K, Akai A and Yamamoto K: Outcomes in patients with critical limb ischemia due to arteriosclerosis obliterans who did not undergo arterial reconstruction. Int Heart J. 59:1041–1046. 2018. View Article : Google Scholar : PubMed/NCBI

2 

Nau JY: Arteriosclerosis obliterans of lower limbs: Early diagnosis before symptoms. Rev Med Suisse. 11:1458–1459. 2015.(In French). PubMed/NCBI

3 

Zhu Y, Xian X, Wang Z, Bi Y, Chen Q, Han X, Tang D and Chen R: Research progress on the relationship between atherosclerosis and inflammation. Biomolecules. 8(pii): E802018. View Article : Google Scholar : PubMed/NCBI

4 

Psychogios K, Stathopoulos P, Takis K, Vemmou A, Manios E, Spegos K and Vemmos K: The pathophysiological mechanism is an independent predictor of Long-Term outcome in stroke patients with large vessel atherosclerosis. J Stroke Cerebrovasc Dis. 24:2580–2587. 2015. View Article : Google Scholar : PubMed/NCBI

5 

Miyahara T and Shigematsu K: Epidemiology, etiology, pathology, and pathophysiology of arteriosclerosis obliterans. Nihon Rinsho. 74 (Suppl 2):S324–S327. 2016.(In Japanese).

6 

He XM, Zheng YQ, Liu SZ, Liu Y, He YZ and Zhou XY: Altered Plasma MicroRNAs as novel biomarkers for arteriosclerosis obliterans. J Atheroscler Thromb. 23:196–206. 2016. View Article : Google Scholar : PubMed/NCBI

7 

Steffens S and Pacher P: The activated endocannabinoid system in atherosclerosis: Driving force or protective mechanism? Curr Drug Targets. 16:334–341. 2015. View Article : Google Scholar : PubMed/NCBI

8 

Yu XH, Zheng XL and Tang CK: Nuclear Factor-kappaB activation as a pathological mechanism of lipid metabolism and atherosclerosis. Adv Clin Chem. 70:1–30. 2015. View Article : Google Scholar : PubMed/NCBI

9 

Yang L, Chu Y, Wang L, Wang Y, Zhao X, He W, Zhang P, Yang X, Liu X, Tian L, et al: Overexpression of CRY1 protects against the development of atherosclerosis via the TLR/NF-kappaB pathway. Int Immunopharmacol. 28:525–530. 2015. View Article : Google Scholar : PubMed/NCBI

10 

Luo H, Wang J, Qiao C, Ma N, Liu D and Zhang W: Pycnogenol attenuates atherosclerosis by regulating lipid metabolism through the TLR4-NF-kappaB pathway. Exp Mol Med. 47:e1912015. View Article : Google Scholar : PubMed/NCBI

11 

Liu R, Fan B, Cong H, Ikuyama S, Guan H and Gu J: Pycnogenol Reduces Toll-Like receptor 4 signaling pathway-mediated atherosclerosis formation in Apolipoprotein E-Deficient mice. J Cardiovasc Pharmacol. 68:292–303. 2016. View Article : Google Scholar : PubMed/NCBI

12 

Kutikhin AG, Ponasenko AV, Khutornaya MV, Yuzhalin AE, Zhidkova II, Salakhov RR, Golovkin AS, Barbarash OL and Barbarash LS: Association of TLR and TREM-1 gene polymorphisms with atherosclerosis severity in a Russian population. Meta Gene. 9:76–89. 2016. View Article : Google Scholar : PubMed/NCBI

13 

Pan S, Lei L, Chen S, Li H and Yan F: Rosiglitazone impedes Porphyromonas gingivalis-accelerated atherosclerosis by downregulating the TLR/NF-kappaB signaling pathway in atherosclerotic mice. Int Immunopharmacol. 23:701–708. 2014. View Article : Google Scholar : PubMed/NCBI

14 

Bhaskar S, Sudhakaran PR and Helen A: Quercetin attenuates atherosclerotic inflammation and adhesion molecule expression by modulating TLR-NF-kB signaling pathway. Cell Immunol. 310:131–140. 2016. View Article : Google Scholar : PubMed/NCBI

15 

Schnittker D, Kwofie K, Ashkar A, Trigatti B and Richards CD: Oncostatin M and TLR-4 ligand synergize to induce MCP-1, IL-6, and VEGF in human aortic adventitial fibroblasts and smooth muscle cells. Mediators Inflamm. 2013:3175032013. View Article : Google Scholar : PubMed/NCBI

16 

Lin J, Kakkar V and Lu X: Impact of MCP-1 in atherosclerosis. Curr Pharm Des. 20:4580–4588. 2014. View Article : Google Scholar : PubMed/NCBI

17 

Chen PY, Qin L, Li G, Tellides G and Simons M: Smooth muscle FGF/TGFβ cross talk regulates atherosclerosis progression. EMBO Mol Med. 8:712–728. 2016. View Article : Google Scholar : PubMed/NCBI

18 

Nikfar S, Saiyarsarai P, Tigabu BM and Abdollahi M: Efficacy and safety of interleukin-1 antagonists in rheumatoid arthritis: A systematic review and Meta-analysis. Rheumatol Int. 38:1363–1383. 2018. View Article : Google Scholar : PubMed/NCBI

19 

Mitoma H, Horiuchi T, Tsukamoto H and Ueda N: Molecular mechanisms of action of anti-TNF-α agents-Comparison among therapeutic TNF-α antagonists. Cytokine. 101:56–63. 2018. View Article : Google Scholar : PubMed/NCBI

20 

Nicoletti F, Di Marco R, Patti F, Reggio E, Nicoletti A, Zaccone P, Stivala F, Meroni PL and Reggio A: Blood levels of transforming growth factor-beta 1 (TGF-beta1) are elevated in both relapsing remitting and chronic progressive multiple sclerosis (MS) patients and are further augmented by treatment with interferon-beta 1b (IFN-beta1b). Clin Exp Immunol. 113:96–99. 1998. View Article : Google Scholar : PubMed/NCBI

21 

Kasashima S, Kawashima A, Endo M, Matsumoto Y, Kasashima F, Zen Y and Nakanuma Y: A clinicopathologic study of immunoglobulin G4-related disease of the femoral and popliteal arteries in the spectrum of immunoglobulin G4-related periarteritis. J Vasc Surg. 57:816–822. 2013. View Article : Google Scholar : PubMed/NCBI

22 

Antoniou S: Rivaroxaban for the treatment and prevention of thromboembolic disease. J Pharm Pharmacol. 67:1119–1132. 2015. View Article : Google Scholar : PubMed/NCBI

23 

Hashikata T, Yamaoka-Tojo M, Namba S, Kitasato L, Kameda R, Murakami M, Niwano H, Shimohama T, Tojo T and Ako J: Rivaroxaban inhibits Angiotensin II-induced activation in cultured mouse cardiac fibroblasts through the modulation of NF-kB pathway. Int Heart J. 56:544–550. 2015. View Article : Google Scholar : PubMed/NCBI

24 

Flierl U, Fraccarollo D, Micka J, Bauersachs J and Schafer A: The direct factor Xa inhibitor Rivaroxaban reduces platelet activation in congestive heart failure. Pharmacol Res. 74:49–55. 2013. View Article : Google Scholar : PubMed/NCBI

25 

Liang Y, Nie H, Ren H, Li F, Tian C, Li H and Zheng Y: Change of Serum Angiopoietin-like Protein 2 and its significance in patients with Arteriosclerotic occlusion. Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 39:188–195. 2017.PubMed/NCBI

26 

Wang SM and Yao C: Standardize the endovascular treatment for arteriosclerosis obliterans. Zhonghua Wai Ke Za Zhi. 54:564–567. 2016.(In Chinese). PubMed/NCBI

27 

Hartman J and Frishman WH: Inflammation and atherosclerosis: A review of the role of interleukin-6 in the development of atherosclerosis and the potential for targeted drug therapy. Cardiol Rev. 22:147–151. 2014. View Article : Google Scholar : PubMed/NCBI

28 

Chen X, Cui R, Li R, Lin H, Huang Z and Lin L: Development of pristane induced mice model for lupus with atherosclerosis and analysis of TLR expression. Clin Exp Rheumatol. 34:600–608. 2016.PubMed/NCBI

29 

Zhong K: Curcumin Mediates a protective effect Via TLR-4/NF-kB signaling pathway in rat model of severe acute pancreatitis. Cell Biochem Biophys. 73:175–180. 2015. View Article : Google Scholar : PubMed/NCBI

30 

Roshan MH, Tambo A and Pace NP: The role of TLR2, TLR4, and TLR9 in the pathogenesis of atherosclerosis. Int J Inflam. 2016:15328322016. View Article : Google Scholar : PubMed/NCBI

31 

Tang YL, Jiang JH, Wang S, Liu Z, Tang XQ, Peng J, Yang YZ and Gu HF: TLR4/NF-kB signaling contributes to chronic unpredictable mild stress-induced atherosclerosis in ApoE-/-mice. PLoS One. 10:e1236852015.

32 

Xie X, Shi X and Liu M: The Roles of TLR gene polymorphisms in atherosclerosis: A systematic review and Meta-Analysis of 35,317 subjects. Scand J Immunol. 86:50–58. 2017. View Article : Google Scholar : PubMed/NCBI

33 

Xu ZR, Li JY, Dong XW, Tan ZJ, Wu WZ, Xie QM and Yang YM: Apple polyphenols decrease atherosclerosis and hepatic steatosis in ApoE-/- Mice through the ROS/MAPK/NF-kB pathway. Nutrients. 7:7085–7105. 2015. View Article : Google Scholar : PubMed/NCBI

34 

Wang X, Chen Q, Pu H, Wei Q, Duan M, Zhang C, Jiang T, Shou X, Zhang J and Yang Y: Adiponectin improves NF-kappaB-mediated inflammation and abates atherosclerosis progression in apolipoprotein E-deficient mice. Lipids Health Dis. 15:332016. View Article : Google Scholar : PubMed/NCBI

35 

Wei M, Li Z, Xiao L and Yang Z: Effects of ROS-relative NF-kB signaling on high glucose-induced TLR4 and MCP-1 expression in podocyte injury. Mol Immunol. 68:261–271. 2015. View Article : Google Scholar : PubMed/NCBI

36 

Imano H, Kato R, Tanikawa S, Yoshimura F, Nomura A, Ijiri Y, Yamaguchi T, Izumi Y, Yoshiyama M and Hayashi T: Factor Xa inhibition by rivaroxaban attenuates cardiac remodeling due to intermittent hypoxia. J Pharmacol Sci. 137:274–282. 2018. View Article : Google Scholar : PubMed/NCBI

37 

Ma J, Li X, Wang Y, Yang Z and Luo J: Rivaroxaban attenuates thrombosis by targeting the NF-kB signaling pathway in a rat model of deep venous thrombus. Int J Mol Med. 40:1869–1880. 2017.PubMed/NCBI

38 

Zuo P, Zuo Z, Wang X, Chen L, Zheng Y, Ma G and Zhou Q: Factor Xa induces pro-inflammatory cytokine expression in RAW 264.7 macrophages via protease-activated receptor-2 activation. Am J Transl Res. 7:2326–2334. 2015.PubMed/NCBI

39 

Han Y, Gao C, Qin B, Xu H, Song X, Li B, Peng B, Fan T and Cheng Z: The effect of anticoagulant therapy on coagulation and inflammation markers in sepsis patients and its significance. Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 27:102–105. 2015.(In Chinese). PubMed/NCBI

40 

Lee JC, Menacherry S, Diehl MC, Giffear MD, White CJ, Juba R, Bagarazzi ML, Muthumani K, Boyer J, Agarwal V, et al: Safety, bioavailability, and pharmacokinetics of VGX-1027-A novel oral anti-inflammatory drug in healthy human subjects. Clin Pharmacol Drug Dev. 5:91–101. 2016. View Article : Google Scholar : PubMed/NCBI

41 

Fagone P, Muthumani K, Mangano K, Magro G, Meroni PL, Kim JJ, Sardesai NY, Weiner DB and Nicoletti F: VGX-1027 modulates genes involved in lipopolysaccharide-induced Toll-like receptor 4 activation and in a murine model of systemic lupus erythematosus. Immunology. 142:594–602. 2014. View Article : Google Scholar : PubMed/NCBI

42 

Stojanovic I, Cuzzocrea S, Mangano K, Mazzon E, Miljkovic D. Wang M, Donia M, Al Abed Y, Kim J, Nicoletti F, et al: In vitro, ex vivo and in vivo immunopharmacological activities of the isoxazoline compound VGX-1027: Modulation of cytokine synthesis and prevention of both organ-specific and systemic autoimmune diseases in murine models. Clin Immunol. 123:311–323. 2007. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

September 2019
Volume 18 Issue 3

Print ISSN: 1792-0981
Online ISSN:1792-1015

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Lou, X., Yu, Z., Yang, X., & Chen, J. (2019). Protective effect of rivaroxaban on arteriosclerosis obliterans in rats through modulation of the toll‑like receptor 4/NF‑κB signaling pathway. Experimental and Therapeutic Medicine, 18, 1619-1626. https://doi.org/10.3892/etm.2019.7726
MLA
Lou, X., Yu, Z., Yang, X., Chen, J."Protective effect of rivaroxaban on arteriosclerosis obliterans in rats through modulation of the toll‑like receptor 4/NF‑κB signaling pathway". Experimental and Therapeutic Medicine 18.3 (2019): 1619-1626.
Chicago
Lou, X., Yu, Z., Yang, X., Chen, J."Protective effect of rivaroxaban on arteriosclerosis obliterans in rats through modulation of the toll‑like receptor 4/NF‑κB signaling pathway". Experimental and Therapeutic Medicine 18, no. 3 (2019): 1619-1626. https://doi.org/10.3892/etm.2019.7726