Skip to main content
NCBI home page
American Journal of Hypertension logoLink to American Journal of Hypertension
. 2023 Nov 20;37(5):349–357. doi: 10.1093/ajh/hpad107

Neutrophil Elastase Inhibition by Sivelestat (ONO-5046) Attenuates AngII-Induced Abdominal Aortic Aneurysms in Apolipoprotein E-Deficient Mice

Yoshiko Hada 1,2, Haruhito A Uchida 3,4,✉, Shugo Okamoto 5, Nozomu Otaka 6,7, Katsuyoshi Katayama 8, Venkateswaran Subramanian 9, Alan Daugherty 10,11, Jun Wada 12
PMCID: PMC13017342  PMID: 37982444

Abstract

BACKGROUND

Abdominal aortic aneurysm (AAA) is an arterial disease characterized by dilatation of the aortic wall. It has been suggested that neutrophil counts and neutrophil elastase activity are associated with AAA. We investigated whether a neutrophil elastase (NE) inhibitor, sivelestat (Siv), had a protective effect against angiotensin II (AngII)-induced AAAs.

METHODS

Male apolipoprotein E-deficient mice were assigned into three groups: Vehicle + saline, AngII + saline, and AngII + Siv. All mice were administered intraperitoneally with either Siv or vehicle twice daily after AngII infusion.

RESULTS

In the 4-week AngII infusion study, plasma NE concentration (P = 0.041) and its activity (P = 0.011) were elevated by AngII. These increases were attenuated by Siv (concentration:P = 0.010, activity:P = 0.027). Further, plasma elastase activity was closely correlated with aortic width (R = 0.6976, P < 0.001). In the 1-week AngII infusion study, plasma and tissue elastase activity increased by AngII (plasma:P = 0.034, tissue:P < 0.001), but were reduced by Siv (plasma:P = 0.014, tissue:P = 0.024). AngII increased aortic width (P = 0.011) but was attenuated by co-administration of Siv (P = 0.022). Moreover, Siv decreased the incidence of AAAs (P = 0.009). Elastin fragmentation induced by AngII was reduced by Siv. Many inflammatory cells that were either CD68 or Gr-1 positive were observed in the AngII + saline group, whereas few inflammatory cells were accumulated in the AngII + Siv group. MMP-2 and MMP-9 were enhanced by AngII, but were reduced by Siv. In vitro, MMP-2 activity was induced by human NE (medium:P < 0.001, cells:P = 0.001), which was attenuated by co-incubation of Siv in medium (P < 0.001) and protein of human aortic smooth muscle cells (P = 0.001).

CONCLUSIONS

Siv attenuated AngII-induced AAA through the inhibition of NE.

Keywords: abdominal aortic aneurysm, blood pressure, elastase, human aortic smooth muscle cells, hypertension, sivelestat

Graphical Abstract

Graphical Abstract.

Graphical Abstract

Open in a new tab

Abdominal aortic aneurysm (AAA) is the progressive dilatation of the abdominal artery. AAA is commonly defined when the maximal abdominal aortic width exceeds 3.0 cm. The risk of aneurysm rupture increases in aortic-width dependent manner.1 Most AAA patients are asymptomatic until rupture, although a few AAAs have vague symptoms. The most cause of death for AAAs is aneurysm rupture, which accounts for approximately 65% of mortality.2 Among people over 60 years of age, the estimated prevalence of AAA in men is 4–8%, compared to 0.5–1.5% in women.3 The major risk factors of AAA include cigarette smoking, aging, male gender, and corresponding family history.4 The incidence of AAA has increased in recent years, due to the aging population, a large number of smokers by population growth especially in the developing countries, the introduction of screening programs, and improved diagnostic methods.5

The pathogenesis of AAA is complex. Several studies report the pathophysiology of AAA is a multifactorial process consisting of inflammation, oxidative stress, matrix metalloproteinase (MMP) activation, intraluminal thrombus, smooth muscle apoptosis, and extracellular matrix degeneration.5

Although neutrophil infiltration is commonly an initial response to tissue injury, neutrophil deficiency alleviated AAA.6 In addition, a cohort study showed a strong correlation between elevated neutrophil counts and AAAs.7 Therefore, neutrophils play an important role in AAA development. Previous studies demonstrated that elastase activity was associated with aneurysm size.8 In addition, it is well known that AAA can be experimentally induced by elastase treatment or AngII administration. Recent research indicated that CD11c + depletion attenuated AngII-induced abdominal aortic matrix degradation indicated by decreased neutrophil elastase (NE) activity and lower elastin degradation score in apolipoprotein E-deficient (apoE−/−) mice.9 Elastase activity in the aorta may determine aneurysm formation. However, little has been reported about the improvement of AAA by elastase attenuation in experimental models.

Sivelestat sodium hydrate (ONO-5046, Siv) is a specific NE inhibitor currently used for treatment of acute lung injury and acute respiratory distress syndrome.10 Many papers have reported that Siv protects lung, liver, heart, kidney, nerve tissue, spinal cord tissue, and other organs from functional injury.11 As with acute lung injury/acute respiratory distress syndrome, AAA is developed by direct or indirect tissue damage by neutrophil activation and secretion of cytotoxic molecules including NE.12 More recent study has indicated that administration of Siv attenuated endotoxin-induced neutrophil activity in vitro.13 However, few studies have demonstrated the role of NE on AAA development.

So far, no pharmacologic treatment has been established to prevent AAA development by inhibiting elastase and its activity. Few studies have focused on the effects of elastase inhibitors on AAA. In this study, we aimed to determine the effects of Siv on AngII-induced AAA in apoE−/− mice.

METHODS

Animals and sivelestat

Male, 8–10 weeks old, apoE−/− mice were purchased from The Jackson Laboratory (Bar Harbor, Cat# 2052). All mice were maintained in a barrier facility, and ambient temperature ranged from 20 to 24 °C. Mice were normal laboratory rodent diet and water ad libitum. The experimental protocol was approved by the Ethics Review Committees for Animal Experimentation of Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical sciences (OKU-2022575). Siv was a generous gift from Ono Pharmaceutical CO., LTD.

Experimental design

The study mice were assigned into three groups: vehicle + saline (n = 8), AngII + saline (n = 14), and AngII + Siv (n = 17). For aneurysm quantification and histological analysis, vehicle or AngII (1,000 ng/min/kg, Bachem, Cat. No. H-1705) was infused via Alzet mini-osmotic pumps (Durect Corporation, Model 2004) for 28 days. Under anesthesia with isoflurane, mini-osmotic pumps were implanted subcutaneously on the right flank, as described previously.14 All mice were administered intraperitoneally with either Siv (100 mg/kg) or vehicle twice daily after the pump implantation for 28 days or 1 week. Tissues and plasma were harvested from mice under anesthetizing with isoflurane (1.5–3.0%, flow rate of 200–300 mL/min, inhalation).

Blood pressure measurement

Systolic blood pressure was measured by sphygmomanometry using a tail cuff system (Visitech systems, BP-2000) following a published protocol. Conscious mice were introduced into a small holder mounted on a thermostatically controlled warming plate and maintained at 37 °C during measurement.

Total cholesterol measurement

Total cholesterol was determined in individual plasma samples using commercially available enzymatic-based kits (Wako Chemicals, Cat# 439–17501).

Neutrophil elastase concentrations activity measurements

Elastase concentration was determined in individual plasma samples using a commercially available quantitative sandwich enzyme immunoassay kits (R&D Systems, Cat# MELA20). Elastase activity was determined by a method using the highly NE specific synthetic substrate N-methoxysuccinyl-Ala-Ala-Pro-Val p-nitroanilide.15 Briefly, samples were incubated in 0.1 M Tris-HCl buffer (pH 8.0) containing 0.5 M NaCl and 1 mM substrate for 24 hours at 37 °C. After incubation, p-nitroaniline was measured spectrophotometrically at 405 nm and was considered to be a measure of NE activity.

Quantification of aneurysms

After 28 days of either saline or AngII infusion, mouse aortas were harvested for aneurysm quantification. Aortas were perfused with saline by left ventricular puncture and were fixed in 10% formalin overnight. Adventitial fat was removed, and the maximum external width of the suprarenal aorta was measured using computerized morphometry (Olympus, cellSens Ver.1.18) as described previously.16 Aneurysm was determined as a 50% increase in aortic width compared to aortas from saline-infused mice. In saline-infused mice, the mean suprarenal width was 0.84 mm; consequently, we defined abdominal AAA as >1.26 mm.

Histological analysis

Mouse abdominal aortas were embedded in optimal cutting temperature compound, and sectioned serially at 10 μm thickness. Verhoeff’s staining was used to evaluate elastin fiber integrity. Immunohistochemical staining was performed to detect macrophage and neutrophil using CD68 antibody (Serotec, Cat# MCA1957), Gr-1 antibody (Invitrogen, Cat# 14-5931-81), and 8-hydroxy-2ʹ-deoxyguanosine (8-OHdG) (Japan Institute for the Control of Aging NIKKEN SEIL, Cat. No. clone N45.1), respectively. Reactivity of the antibodies with tissue antigens was detected using AEC and ImmPACT AEC HRP Substrate (Vector Laboratories) as described previously.17

Cell culture

Human aortic smooth muscle cells (HASMCs) were purchased from Kurabo (Cat# KS-4009, Lot. 01127) and were grown in Smooth Muscle Cell Growth Medium (SmGM 2 BulletKit: Lonza, Cat# CC-3182). All experiments were performed at passage 7 to 8. For cell culture experiments, 1 × 106 cells were preincubated with Siv (100 μM) or DMSO and stimulated with 0.1 μg/mL human NE (Merck, Cat# 324681, Lot.3841832) for 24 hours at six-well plates. Cultured medium and protein lysate from HASMCs were collected for zymography.

Gelatin zymography

The mice infused with saline or AngII were administered intraperitoneally with either Siv (100 mg/kg) or vehicle twice daily, and a week later, the aortas were harvested. Proteins were extracted from aortas without any discernable aneurysms. Samples in vivo or in vitro were resolved under non-reducing condition on a 7.5% polyacrylamide gel containing 0.1% gelatin as a substrate for MMP activity. The bands were quantified using a General Electric Imager (LAS 4000 mini).

Statistics

All statistical analyses were performed using Sigma Plot v14.0 (Systat Software Inc., California, USA). Data are presented as mean ± standard deviation or standard error of the mean where appropriate. Statistical significance among multiple groups was assessed by one-way analysis of variance or one-way analysis of variance on rank, where appropriate, followed by Holm–Sidak post hoc test or Student–Newman–Keuls post hoc test. AAA incidence was analyzed by Fisher’s exact test. Pearson’s correlation coefficients were used for the correlation analysis. A P values <0.05 was considered as statistically significant.

RESULTS

The effect of sivelestat on elastase concentration and its activity in plasma or aortic tissue

In a 4-week AngII infusion study, plasma elastase concentration (4.76 ± 0.67 ng/mL vs. 11.41 ± 2.42 ng/mL, P = 0.041) and activity (1.65 ± 0.71 μM vs. 8.73 ± 2.13 μM, P = 0.011) were increased by AngII. These increases were significantly attenuated by Siv administration (concentration: 5.46 ± 0.76 ng/mL, P = 0.010 vs. AngII + saline, activity: 3.91 ± 0.84 μM, P = 0.027 vs. AngII + saline; Figure 1a,b). A positive correlation was observed between aortic width and plasma elastase activity (R = 0.6976, P < 0.001; Figure 1c). In a 1-week AngII infusion study, plasma elastase activity increased by AngII (1.582 ± 0.216 μM vs. 4.992 ± 1.411 μM, P < 0.034) and reduced by Siv administration (1.608 ± 0.222 μM, P = 0.014 vs. AngII + saline; Figure 1d). In addition, tissue elastase activity was significantly increased with AngII (0.19 ± 0.09 μM vs. 1.04 ± 0.02 μM, P < 0.001), but was reduced by Siv administration (0.78 ± 0.05 μM, P = 0.024 vs. AngII + saline; Figure 1e).

Figure 1.

Figure 1.

Open in a new tab

The effect of sivelestat on neutrophil elastase concentration and activity in plasma or aorta tissue. (a) In the 4-week AngII infusion study, plasma neutrophil elastase concentrations are shown in each group. (b) In the 4-week AngII infusion study, plasma elastase activity was shown in each group (n = 8–17 in each group). Each bar presents the mean + SEM. (c) Scatterplots of aortic width and plasma elastase activity among 4-week AngII infusion study mice (n = 22) were shown. Pearson’s correlation coefficients were used for the correlation analysis. (d) In the 1-week AngII infusion study, plasma elastase activity was shown in each group. (e) In the 1-week AngII infusion study, aorta tissue elastase activity was shown in each group (n = 5 in each group). Each bar presents the mean + SEM. The Vehicle + saline group: blue bar, the AngII + saline group: red bar, and the AngII + Siv group: yellow bar. Statistical analysis was performed by one-way analysis of variance (ANOVA).

Sivelestat had no effect on body weight, blood pressure, and lipid concentrations

No significant differences were found in body weight and plasma concentrations of total cholesterol among the three groups. Although AngII infusion significantly increased systolic blood pressure, no difference was observed between the AngII + saline and AngII + Siv groups (Table 1).

Table 1.

Characteristics of study mice

Vehicle + saline AngII + saline AngII + Siv
n 8 14 17
BW (g) 26.8 ± 2.0 26.5 ± 3.0 26.2 ± 2.2
SBP (mm Hg) 104 ± 11 165 ± 12* 157 ± 14*
T-Cho (mg/dL) 456 ± 65 544 ± 84 493 ± 103
Open in a new tab

Data represent mean ± SD.

Abbreviations: AngII, angiotensin II; BW, body weight; n, number; SBP, systolic blood pressure; Siv, sivelestat; T-Cho, total cholesterol concentration.

*P < 0.05 vs. Vehicle + saline (One-way ANOVA on Rank).

Sivelestat attenuated formation of AngII-induced AAA

To determine the effects of Siv on development of AngII-induced AAAs, the maximum external width of the suprarenal aorta was measured. AngII infusion significantly increased aortic width compared to the saline group (mean width of abdominal aorta: vehicle + saline 0.84 ± 0.03 mm, AngII + saline 1.92 ± 0.17 mm; Figure 2b, representative photo in Figure 2a). Siv administration attenuated dilatation of aortic width and decreased AAA formation (mean aortic width: 1.43 ± 0.12 mm, P = 0.022; Figure 2b, representative photo in Figure 2a). Based on definition of AAA as >50% increase in aortic width, the incidence of AAAs in AngII-infused mice without Siv administration was 93%. Administration of Siv significantly decreased the incidence to 47% (P = 0.009; Figure 2c). To evaluate the histological characteristics of abdominal aortas, Verhoeff’s staining was performed on tissue sections from suprarenal aortas. Pronounced disruptions of medial layers were observed in AngII-infused mice; however, elastin fragmentation induced by AngII infusion were markedly reduced in mice administered with Siv (Figure 2d).

Figure 2.

Figure 2.

Open in a new tab

The effect of sivelestat on AngII-induced abdominal aortic aneurysm (AAA). (a) Representative images of abdominal aortas for each group. Left: Vehicle + saline, center: AngII + saline, and right: AngII + Siv. (b) Aortic diameter measurements from each mouse are represented by a circle ). (● represents vehicle-infused mice without Siv administration, ● represents AngII-infused mice without Siv administration, ● represents AngII-infused mice with Siv administration, ◆ represents means, and—represents SEM. Vehicle + saline (n = 8), AngII + saline (n = 14), AngII + Siv (n = 16). Statistical analysis was performed by one-way analysis of variance (ANOVA) on rank. (c) The incidence of AAA (>50% increase in aortic width) was 93% in the AngII + saline group (red bar) compared to 0% in the Vehicle + saline group and 47% in the AngII + Siv group (yellow bar). Statistical analyses were performed by Fisher Exact test (P = 0.009). (d) Representative images of Verhoeff van Gieson’s staining. The scale bar indicates 200 μm (upper row) and 50 μm (lower row).

Sivelestat reduced the medial disruption induced by AngII

A large number of CD68 positive cells and Gr-1 positive cells were observed in the AngII + saline group. In the Siv administered groups, as well as the vehicle + saline group, few accumulations of inflammatory cells were observed. Thus, Siv appeared to suppress the accumulation of macrophages and neutrophils (Figure 3a,b). AngII increased the number of 8-OHdG positive cells in the aorta, suggesting that DNA damage increased. Treatment with Siv reduced the number of 8-OHdG positive cells in the aorta (Figure 3c).

Figure 3.

Figure 3.

Open in a new tab

The effect of sivelastat on AngII-induced the medial disruption. Immunostainings (a, b) are shown. These sections were stained with mAbs against CD68 (a) and Gr-1 (b), respectively. The scale bar indicates 200 μm (upper row in a and b) and 50 μm (lower row in a and b).

Sivelestat reduced MMP in the aorta, HASMC, and medium

Gelatin zymography was performed to detect MMP-2 and MMP-9. Pro-form MMP-2 (65 kDa and 70 kDa) and active-form MMP-2 (58 kDa) were increased by AngII infusion and reduced by Siv administration (pro-form: P = 0.071, active-form: P = 0.022; Figure 4a). Pro-form MMP-9 (92 kDa) and active-form MMP-9 (82 kDa) were increased by AngII, and only active-form MMP-9 (82 kDa) was reduced by Siv administration (active-form: P = 0.042; Figure 4b). Thus, Siv reduced both the activities of MMP-2 and MMP-9 in the aorta.

Figure 4.

Figure 4.

Open in a new tab

The effect of sivelestat on MMP in the aorta, HASMC, and medium. (a) The relative abundances of pro- or active-form of MMP-2 (pro-form: 65 kDa and 70 kDa, active-form: 58 kDa) were quantified by densitometry and normalized to the abundance under the control condition (n = 5 in each group). Each bar presents the mean + SEM. (b) The relative abundances of pro- or active-form of MMP-9 (pro-form: 92 kDa, active-form: 82 kDa) were quantified by densitometry and normalized to the abundance under the control condition (n = 5 in each group). Each bar presents the mean + SEM. The Vehicle + saline group: blue bar, the AngII + saline group: red bar, and the AngII + Siv group: yellow bar. (c) The relative abundances of pro- and active-form of MMP-2 (pro-form: 65 kDa, active-form: 58 kDa) in HASMC were quantified by densitometry and normalized to the abundance under control conditions (n = 5 in each group). Each bar presents the mean + SEM. (d) The relative abundances of pro- or active-form of MMP-2 in medium were quantified by densitometry and normalized to the abundance under the control condition (n = 5 in each group). Each bar presents the mean + SEM. The Siv(−)/elastase(−) group: blue bar, the Siv(−)/elastase(+) group: red bar, and the Siv(+)/elastase(+) group: yellow bar. Statistical analysis was performed by one-way analysis of variance (ANOVA).

Active-form MMP-2 (58 kDa) in HASMC was increased by elastase and reduced by Siv treatment (active-form: P = 0.001; Figure 4c), although pro-form MMP-2 (65 kDa) showed no significant difference among three groups. Similarly, active-form MMP-2 (58 kDa) in medium were increased by elastase and reduced by Siv treatment (active-form: P < 0.001; Figure 4d), although pro-form MMP-2 (65 kDa) had no significant difference between with and without elastase. Thus, Siv reduced the activities of MMP-2 in both HASMC and medium.

DISCUSSION

Siv is a specific NE inhibitor and has been used to treat acute lung injury and acute respiratory distress syndrome, however, it remains unknown whether the drug can exert any beneficial effects on AAAs. Our results demonstrated that Siv attenuated AngII-induced AAA development into apoE−/− mice via reducing elastase activity.

The proteolytic degradation of the aortic wall is a major mechanism in the pathogenesis and progression of AAA. In fact, several proteases, such as NE, MMP-9, and MMP-2, are increased in AAA.18 A few studies, both clinical and animal, suggest that circulating neutrophils may be an important contributor to AAA formation in the early phase.6,7 A cohort study showed that strong associations were observed between elevated neutrophil counts and AAA.7 The current study demonstrated that elastase activity was correlated with aneurysm size. This results support the results of Busuttil et al.8 The elastase-induced AAA model is widely used since perfusion of the aorta with elastase results in aneurysm formation.19 Moreover, neutrophil depletion6 or prevention of their recruitment20 has inhibited experimental AAA formation. Thus, a specific NE inhibitor, Siv, may have the therapeutic potential of AAA in animal models. It is reported that Siv administration attenuated systemic and pulmonary NE activity and decreased neutrophil infiltration and inflammatory cytokine expression in the lung.21 Similarly, in our study, Siv administration attenuated NE activity in plasma and vascular tissue, and decreased neutrophil infiltration in the vascular tissue. Moreover, NE not only degrades various components of the extracellular matrix, but also participates in other important mechanisms of AAA development, such as MMP activation and impaired cellular recolonization of AAA thrombi.22

The role of MMP in AAA development has been demonstrated in animal models.23 In addition, various MMPs have been invoked in human aneurysmal pathology.24 Especially, a positive correlation was reported between MMP-9 or MMP-2 and aortic width.25 In our study, treatment of Siv attenuated NE activation, leading to decrease of infiltration of neutrophils and macrophages at the aneurysmal site. Consequently, MMP-9 activity reduced, resulting in attenuation of matrix degradation. Moreover, in our in vitro study, incubation with elastase enhanced MMP-2 activity, which was attenuated by Siv. In addition, in vivo study showed Siv attenuated MMP-2 activity in the aorta. Given these results, Siv might attenuate the formation of AAA by regulation of MMPs.

NE plays an important role in host defense for neutrophils.26 In this point, it is concerned whether inhibition of NE activity might disrupt the host defense system, resulting in aggravation of infection. However, Siv preserved the host immune defense system,27 and did not influence the bactericidal capacity of neutrophils in vitro.28 In addition, available clinical study data, including for the STRIVE study and the related post-marketing study, show no particular concerns regarding adverse events.10 Recent studies have indicated that Siv suppressed oxidative stress in the lung, dental pulp and other organs.29,30 In line with these studies, Siv reduced the number of 8-OHdG positive cells induced by AngII in the current study, suggesting that Siv attenuated oxidative stress.

In conclusion, our results may bring a great promise in the treatment strategy for AAA. Since Siv is already in clinical use and has proven to be safe, a further large clinical trial should be required to examine the therapeutic effect of Siv on the development and progression of AAA in humans.

Acknowledgments

We thank Dr Yasuhiro Onishi, Dr Mariko Tsuchida-Nishiwaki, and Dr Natsumi Matsuoka-Uchiyama for her technical assistance. We would like to thank Dr Hidemi Takeuchi and Dr Ryoko Umebayashi for his helpful advice to perform several experiments. We thank the Ono Pharmaceutical Co., Ltd for their generous gift of Siv. The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

Contributor Information

Yoshiko Hada, Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry, and Pharmaceutical Science, Okayama, Japan; Department of Chronic Kidney Disease and Cardiovascular Disease, Okayama University Faculty of Medicine, Dentistry, and Pharmaceutical Science, Okayama, Japan.

Haruhito A Uchida, Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry, and Pharmaceutical Science, Okayama, Japan; Department of Chronic Kidney Disease and Cardiovascular Disease, Okayama University Faculty of Medicine, Dentistry, and Pharmaceutical Science, Okayama, Japan.

Shugo Okamoto, Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry, and Pharmaceutical Science, Okayama, Japan.

Nozomu Otaka, Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry, and Pharmaceutical Science, Okayama, Japan; Department of Nephrology and Rheumatology, Kagawa Prefectural Central Hospital, Kagawa, Japan.

Katsuyoshi Katayama, Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry, and Pharmaceutical Science, Okayama, Japan.

Venkateswaran Subramanian, Division of Cardiovascular Medicine, Department of Medicine, University of Missouri, Columbia, Missouri, USA.

Alan Daugherty, Saha Cardiovascular Research Center, University of Kentucky, Lexington, Kentucky, USA; Department of Physiology, University of Kentucky, Lexington, Kentucky, USA.

Jun Wada, Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Faculty of Medicine, Dentistry, and Pharmaceutical Science, Okayama, Japan.

Conflict of interest

The authors have no competing interests to declare that are relevant to the content of this article.

REFERENCES

  • 1. Brown  LC, Powell  JT.  Risk factors for aneurysm rupture in patients kept under ultrasound surveillance UK Small Aneurysm Trial Participants. Ann Surg  1999; 230:289–96; discussion 296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2. Arnaoutakis  DJ, Upchurch  GR  Jr. Abdominal aortic aneurysm screening is safe yet lacks effectiveness. Circulation  2019; 139:1381–1383. [DOI] [PubMed] [Google Scholar]
  • 3. Klink  A, Hyafil  F, Rudd  J, Faries  P, Fuster  V, Mallat  Z, Meilhac  O, Mulder  WJ, Michel  JB, Ramirez  F, Storm  G, Thompson  R, Turnbull  IC, Egido  J, Martín-Ventura  JL, Zaragoza  C, Letourneur  D, Fayad  ZA.  Diagnostic and therapeutic strategies for small abdominal aortic aneurysms. Nat Rev Cardiol  2011; 8:338–347. [DOI] [PubMed] [Google Scholar]
  • 4. Lindquist Liljeqvist  M, Hultgren  R, Bergman  O, Villard  C, Kronqvist  M, Eriksson  P, Roy  J.  Tunica-specific transcriptome of abdominal aortic aneurysm and the effect of intraluminal thrombus, smoking, and diameter growth rate. Arterioscler Thromb Vasc Biol  2020; 40:2700–2713. [DOI] [PubMed] [Google Scholar]
  • 5. Sakalihasan  N, Limet  R, Defawe  OD.  Abdominal aortic aneurysm. Lancet  2005; 365:1577–1589. [DOI] [PubMed] [Google Scholar]
  • 6. Eliason  JL, Hannawa  KK, Ailawadi  G, Sinha  I, Ford  JW, Deogracias  MP, Roelofs  KJ, Woodrum  DT, Ennis  TL, Henke  PK, Stanley  JC, Thompson  RW, Upchurch  GR  Jr. Neutrophil depletion inhibits experimental abdominal aortic aneurysm formation. Circulation  2005; 112:232–240. [DOI] [PubMed] [Google Scholar]
  • 7. Shah  AD, Denaxas  S, Nicholas  O, Hingorani  AD, Hemingway  H.  Neutrophil counts and initial presentation of 12 cardiovascular diseases: a CALIBER Cohort Study. J Am Coll Cardiol  2017; 69:1160–1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8. Busuttil  RW, Rinderbriecht  H, Flesher  A, Carmack  C.  Elastase activity: the role of elastase in aortic aneurysm formation. J Surg Res  1982; 32:214–217. [DOI] [PubMed] [Google Scholar]
  • 9. Krishna  SM, Moran  CS, Jose  RJ, Lazzaroni  S, Huynh  P, Golledge  J.  Depletion of CD11c+ dendritic cells in apolipoprotein E-deficient mice limits angiotensin II-induced abdominal aortic aneurysm formation and growth. Clin Sci (Lond)  2019; 133:2203–2215. [DOI] [PubMed] [Google Scholar]
  • 10. Aikawa  N, Ishizaka  A, Hirasawa  H, Shimazaki  S, Yamamoto  Y, Sugimoto  H, Shinozaki  M, Taenaka  N, Endo  S, Ikeda  T, Kawasaki  Y.  Reevaluation of the efficacy and safety of the neutrophil elastase inhibitor, sivelestat, for the treatment of acute lung injury associated with systemic inflammatory response syndrome; a phase IV study. Pulm Pharmacol Ther  2011; 24:549–554. [DOI] [PubMed] [Google Scholar]
  • 11. Nishiyama  J, Matsuda  M, Ando  S, Hirasawa  M, Suzuki  T, Makuuchi  H.  The effects of the early administration of sivelestat sodium, a selective neutrophil elastase inhibitor, on the postoperative course after radical surgery for esophageal cancer. Surg Today  2012; 42:659–665. [DOI] [PubMed] [Google Scholar]
  • 12. Estrada  S, Elgland  M, Selvaraju  RK, Mani  K, Tegler  G, Wanhainen  A, Wågsäter  D, Bergström  M, Jimenez-Royo  P, Jahan  M, Nordeman  P, Antoni  G.  Preclinical evaluation of [(11)C]GW457427 as a tracer for neutrophil elastase. Nucl Med Biol  2022; 106–107:62–71. [DOI] [PubMed] [Google Scholar]
  • 13. Okeke  EB, Louttit  C, Fry  C, Najafabadi  AH, Han  K, Nemzek  J, Moon  JJ.  Inhibition of neutrophil elastase prevents neutrophil extracellular trap formation and rescues mice from endotoxic shock. Biomaterials  2020; 238:119836. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14. Umebayashi  R, Uchida  HA, Wada  J.  Abdominal aortic aneurysm in aged population. Aging (Albany NY)  2018; 10:3650–3651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15. Yoshimura  K, Nakagawa  S, Koyama  S, Kobayashi  T, Homma  T.  Roles of neutrophil elastase and superoxide anion in leukotriene B4-induced lung injury in rabbit. J Appl Physiol (1985)  1994; 76:91–96. [DOI] [PubMed] [Google Scholar]
  • 16. Otaka  N, Uchida  HA, Okuyama  M, Hada  Y, Onishi  Y, Kakio  Y, Takeuchi  H, Umebayashi  R, Tanabe  K, Subramanian  V, Daugherty  A, Sato  Y, Wada  J.  Vasohibin-2 aggravates development of ascending aortic aneurysms but not abdominal aortic aneurysms nor atherosclerosis in ApoE-deficient mice. Am J Hypertens  2021; 34:467–475. [DOI] [PubMed] [Google Scholar]
  • 17. Hada  Y, Uchida  HA, Mukai  T, Kojima  F, Yoshida  M, Takeuchi  H, Kakio  Y, Otaka  N, Morita  Y, Wada  J.  Inhibition of interleukin-6 signaling attenuates aortitis, left ventricular hypertrophy and arthritis in interleukin-1 receptor antagonist deficient mice. Clin Sci (Lond)  2020; 134:2771–2787. [DOI] [PubMed] [Google Scholar]
  • 18. Riches  K, Angelini  TG, Mudhar  GS, Kaye  J, Clark  E, Bailey  MA, Sohrabi  S, Korossis  S, Walker  PG, Scott  DJ, Porter  KE.  Exploring smooth muscle phenotype and function in a bioreactor model of abdominal aortic aneurysm. J Transl Med  2013; 11:208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. Chew  DK, Orshal  JM, Khalil  RA.  Elastase-induced suppression of endothelin-mediated Ca2+ entry mechanisms of vascular contraction. Hypertension  2003; 42:818–824. [DOI] [PubMed] [Google Scholar]
  • 20. Pagano  MB, Bartoli  MA, Ennis  TL, Mao  D, Simmons  PM, Thompson  RW, Pham  CT.  Critical role of dipeptidyl peptidase I in neutrophil recruitment during the development of experimental abdominal aortic aneurysms. Proc Natl Acad Sci USA  2007; 104:2855–2860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. Ishii  T, Doi  K, Okamoto  K, Imamura  M, Dohi  M, Yamamoto  K, Fujita  T, Noiri  E.  Neutrophil elastase contributes to acute lung injury induced by bilateral nephrectomy. Am J Pathol  2010; 177:1665–1673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22. Fontaine  V, Touat  Z, Mtairag el  M, Vranckx  R, Louedec  L, Houard  X, Andreassian  B, Sebbag  U, Palombi  T, Jacob  MP, Meilhac  O, Michel  JB.  Role of leukocyte elastase in preventing cellular re-colonization of the mural thrombus. Am J Pathol  2004; 164:2077–2087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. Longo  GM, Xiong  W, Greiner  TC, Zhao  Y, Fiotti  N, Baxter  BT.  Matrix metalloproteinases 2 and 9 work in concert to produce aortic aneurysms. J Clin Invest  2002; 110:625–632. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24. Hobeika  MJ, Thompson  RW, Muhs  BE, Brooks  PC, Gagne  PJ.  Matrix metalloproteinases in peripheral vascular disease. J Vasc Surg  2007; 45:849–857. [DOI] [PubMed] [Google Scholar]
  • 25. Coutard  M, Touat  Z, Houard  X, Leclercq  A, Michel  JB.  Thrombus versus wall biological activities in experimental aortic aneurysms. J Vasc Res  2010; 47:355–366. [DOI] [PubMed] [Google Scholar]
  • 26. Belaaouaj  A, McCarthy  R, Baumann  M, Gao  Z, Ley  TJ, Abraham  SN, Shapiro  SD.  Mice lacking neutrophil elastase reveal impaired host defense against gram negative bacterial sepsis. Nat Med  1998; 4:615–618. [DOI] [PubMed] [Google Scholar]
  • 27. Hagio  T, Kishikawa  K, Kawabata  K, Tasaka  S, Hashimoto  S, Hasegawa  N, Ishizaka  A.  Inhibition of neutrophil elastase reduces lung injury and bacterial count in hamsters. Pulm Pharmacol Ther  2008; 21:884–891. [DOI] [PubMed] [Google Scholar]
  • 28. Zeiher  BG, Matsuoka  S, Kawabata  K, Repine  JE.  Neutrophil elastase and acute lung injury: prospects for sivelestat and other neutrophil elastase inhibitors as therapeutics. Crit Care Med  2002; 30:S281–S287. [DOI] [PubMed] [Google Scholar]
  • 29. Yao  W, Han  X, Guan  Y, Guan  J, Wu  S, Chen  C, Li  H, Hei  Z.  Neutrophil elastase inhibitors suppress oxidative stress in lung during liver transplantation. Oxid Med Cell Longev  2019; 2019:7323986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30. Prakash  R, Mishra  RK, Ahmad  A, Khan  MA, Khan  R, Raza  SS.  Sivelestat-loaded nanostructured lipid carriers modulate oxidative and inflammatory stress in human dental pulp and mesenchymal stem cells subjected to oxygen-glucose deprivation. Mater Sci Eng C Mater Biol Appl  2021; 120:111700. [DOI] [PubMed] [Google Scholar]

Articles from American Journal of Hypertension are provided here courtesy of Oxford University Press

RESOURCES