|Year : 2020 | Volume
| Issue : 2 | Page : 90-94
The expression and significance of serum caveolin-1 in patients with Kawasaki disease
Feng Zhu1, Jing Huang2, Xuliang Wang2, Ping Li2, Yaoyao Yan2, Yunyun Zheng2, Yue'e He2, Tingting Wu2, Yue Ren2, Rongzhou Wu2
1 Department of Child Healthcare, Wenzhou People's Hospital, Wenzhou, Zhejiang, China
2 Children's Heart Center, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Institute of Cardiovascular Development and Translational Medicine, The Second School of Medicine, Wenzhou Medical University, Wenzhou, Zhejiang, China
|Date of Submission||30-Sep-2019|
|Date of Acceptance||25-Feb-2020|
|Date of Web Publication||27-Apr-2020|
Dr. Rongzhou Wu
Children's Heart Center, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Institute of Cardiovascular Development and Translational Medicine, The Second School of Medicine, Wenzhou Medical University, Wenzhou, Zhejiang 325027
Source of Support: Zhejiang Province (Grant no. 2017KY465, Zhejiang, China)
and the Chinese Medicine Scientific Research Foundation
Project of Zhejiang Province (Grant no. 2010ZA092)., Conflict of Interest: None
We investigated the expression of caveolin-1 (Cav-1) in Kawasaki disease (KD) and analyzed its relationship with coronary artery lesions (CALs). Cav-1 participated in the progression of CAL in KD. A total of 68 children with KD (23 with CALs), age matched with a fever control group (F, n = 28) and a normal control group (N, n = 24) were enrolled in this study. Cav-1 expression was detected using an enzyme-linked immunosorbent assay. The results are the following: (1) Compared with the F and N, Cav-1 expression was significantly increased in the children with KD (P < 0.05); there was no significant difference in Cav-1 between the F and N. (2) The serum level of Cav-1 was significantly higher in children with KD and CALs during the acute phase than in children with KD without CALs (P < 0.05). (3) Serum Cav-1 may be a biomarker that reflects CALs in children with KD based on a receiver operating characteristic (ROC) curve analysis. (4) Those children with KD who were given intravenous immunoglobulin (2 g/kg, 10–12 h) during the acute phase showed decreased expression of Cav-1 compared to the N. Conclusions are as follows: (1) The serum level of Cav-1 during the acute phase of KD increased significantly, while in KD patients with CALs the increase was even greater. (2) Based on our ROC curve analysis, Cav-1 may be a predictor of CALs in children with KD.
Keywords: Caveolin-1, coronary artery lesions, intravenous immunoglobulin, Kawasaki disease
|How to cite this article:|
Zhu F, Huang J, Wang X, Li P, Yan Y, Zheng Y, He Y, Wu T, Ren Y, Wu R. The expression and significance of serum caveolin-1 in patients with Kawasaki disease. Chin J Physiol 2020;63:90-4
|How to cite this URL:|
Zhu F, Huang J, Wang X, Li P, Yan Y, Zheng Y, He Y, Wu T, Ren Y, Wu R. The expression and significance of serum caveolin-1 in patients with Kawasaki disease. Chin J Physiol [serial online] 2020 [cited 2020 Dec 2];63:90-4. Available from: https://www.cjphysiology.org/text.asp?2020/63/2/90/283347
| Introduction|| |
Kawasaki disease (KD), or mucocutaneous lymph node syndrome, a multisystem inflammatory condition observed in younger children, is an acute systemic vasculitis that primarily affects small- and medium-sized arteries. In developed countries, KD is the most commonly acquired heart disease in children. Its most serious, complications are coronary artery lesions (CALs), including dilatations and aneurysms, which have been reported in 20%–25% of untreated children.
Caveolin-1 (Cav-1), which is expressed in caveolae membranes, regulates the activity of signal transduction pathways between inflammatory cells and endothelial cells. It also affects the response of inflammatory cells to inflammatory mediators and adjusts the inflammatory reaction. Cav-1 has been studied in chronic vascular inflammatory diseases such as atherosclerosis, diabetic microvascular disease, vascular restenosis, and angiopathic diseases.,,, However, whether Cav-1 is involved in the development of KD is unknown. The purpose of this study was to investigate the clinical value of serum Cav-1 levels as a predictor of CALs in children with KD.
| Materials and Methods|| |
We enrolled 68 KD patients between February 2016 and August 2017 at the Second Affiliated Hospital of Wenzhou Medical College. The diagnosis of KD was based on the 2004 American Heart Association/American Academy of Pediatrics guidelines. Excluding intravenous immunoglobulin (IVIG) treatment prior to admission (nonhospital initial diagnosis and initial treatment), the study patients had either gone through the acute phase (fever 1–2 weeks later) or received a diagnosis at our hospital but had not received IVIG treatment. Echocardiography was used to detect the presence of CALs during the disease. Blood samples were taken at the acute stage in KD patients with a normal temperature on the 3rd day after the completion of the initial IVIG treatment (KD patients during the acute period were administered IVIG [2 g/kg of body weight, per day]).
In addition to the 68 KD patients, 28 children with respiratory tract infections were included as febrile controls. These children did not have a cutaneous eruption or myocardial injury, and they were excluded if they had abnormal myocardial enzyme levels or an abnormal electrocardiogram.
Furthermore, 24 healthy children with a normal medical history, physical examination, and laboratory tests were selected as the normal control group. The study was approved by the Ethics Committee of the Second Affiliated Hospital of Wenzhou Medical University (no. L-2016-07, Date: January 02, 2016). Informed consent was obtained from all participants' parents or legal guardians. There were no significant differences between the groups regarding baseline characteristics.
Measurement of serum Caveolin-1 levels
Blood samples (2 ml for each subject) were taken during the acute stage in KD patients and after IVIG treatment (when subjects had a normal temperature lasting 48–72 h). The samples were centrifuged at 3,000 rpm/min for 10 min and stored at −80°C until analysis. Serum Cav-1 concentrations were measured using a commercially available enzyme-linked immunosorbent assay kit (Shanghai XiTang Biological Technology Co. Ltd., Shanghai, China) according to the manufacturer's instructions. The same method was used to measure Cav-1 levels in the febrile and healthy control groups.
Data analysis involved the use of SPSS 19.0 statistical software (IBM Corp., Armonk, NY, USA) and Excel (Microsoft Corp., Redmond, WA, USA). Data are expressed as the mean ± standard deviation. Student's t-test was used to compare the mean values of different groups. A Chi-squared test was used to compare categorical variables. A receiver operating characteristic (ROC) curve analysis was performed to define the serum Cav-1 concentration at which the sensitivity and specificity were optimal. P < 0.05 was considered to indicate statistical significance.
| Results|| |
Demographic data of the participants
The KD group included 44 boys and 24 girls with a median age of 33.6 ± 2.9 months. Of these, 23 (33.8%) developed CALs based on the echocardiographic results, and 45 (66.2%) did not develop CALs (NCALs). The febrile control group included 19 boys and 9 girls with a median age of 39.1 ± 6.2 months. The normal controls consisted of 13 boys and 11 girls with a median age of 33.1 ± 4.1 months.
Comparison of Caveolin-1 levels in the acute Kawasaki disease group and control groups
The mean serum Cav-1 level during the acute stage in the KD patients (40.32 ± 21.50 ng/ml) was significantly higher than that in the normal control group (16.70 ± 10.80 ng/ml) and the febrile control group (26.92 ± 14.19 ng/ml) (all, P < 0.05). This level was not significantly different between the healthy and febrile children (P > 0.05) [Table 1]. In the KD patients, we did not observe statistical differences in Cav-1 levels between boys (median 37.15 [range 24.45–61.96]) and girls (median 34.49 [range 18.47–47.87]).
Comparison between the coronary artery lesion and not develop coronary artery lesions groups
The mean serum Cav-1 level in the CAL group (51.22 ± 23.24 ng/ml) was significantly higher than that in the NCAL group (34.75 ± 18.44 ng/ml) (P < 0.05) [Table 2].
|Table 2: Comparison of patients with and without coronary artery lesions|
Click here to view
Receiver operating characteristic curves
To assess the performance of the serum level of Cav-1 in predicting CALs, ROC curves were plotted, and the area under the curve (AUC) was calculated. As shown in [Figure 1], the AUC of Cav-1 for predicting CALs was 0.702 (95% confidence interval [CI] 0.565–0.839, P = 0.007). When a cutoff value of 58.14 ng/ml for Cav-1 based on the ROC curve was obtained for predicting CALs, it generated a sensitivity of 91.1% and a specificity of 47.8%.
|Figure 1: Predictive performance of the serum caveolin-1 level. Receiver operating characteristic (ROC) curves of the serum Cav-1 levels used to predict coronary artery lesions in children diagnosed with Kawasaki disease. The area under the curve (AUC) of serum Cav-1 was 0.702|
Click here to view
Comparison of Caveolin-1 before and after intravenous immunoglobulin treatment in Kawasaki disease patients
A comparison of the mean serum Cav-1 levels after IVIG treatment (19.77 ± 10.47 ng/ml) and before IVIG treatment (40.32 ± 21.50 ng/ml) showed a significant difference (P < 0.05) [Table 3].
|Table 3: Comparison of serum caveolin-1 level before and after intravenous immunoglobulin treatment in Kawasaki disease patients|
Click here to view
| Discussion|| |
KD is an acute, nonspecific systemic vascular inflammatory syndrome. A considerable volume of research has shown that the pathological process of KD is characterized by immune activation and immune vasculitis in vascular endothelial cells. KD mostly affects small- and medium-sized arteries, especially the coronary artery, resulting in CALs (e.g., dilation, aneurysm formation, or advanced stenosis). CALs are the main cause of death in children with KD. Research has shown that long-term CALs, including thickening and calcification, result in localized stenosis and myocardial ischemia. Currently, coronary arterial dilatation is an index for CAL. According to the literature, endotheliocytes arise from existing injury or dysfunction even in the normal structure of the coronary artery. Therefore, this index is defective.
Caveolae are small (50–100 nm) pit-like depressions, similar to flask depressions, in the cell membrane. Cav-1, an important functional protein, is widely expressed in endothelial cells, smooth muscle cells, fibroblasts, macrophages, lymphocytes, and neutrophils. Apart from inducing the formation of caveolae and maintaining their structure, Cav-1 participates in various inflammatory signal transduction pathways, regulates the production of cytokines and the release of mediators of the inflammatory response, and regulates the activation of inflammatory cells (neutrophils and macrophages); thus, it regulates inflammation in the body. During the control of the inflammatory signal transduction pathway, Cav-1 activates or represses the activity of various signaling molecules by combining a dimer located in the caveolin scaffolding domain with signaling molecules such as endothelial nitric oxide synthase Cyclooxygenase-2., It then affects the activities of receptors related to signal transduction, kinases, and connexins and regulates inflammation. Moreover, it plays an important role in angiogenesis, cell differentiation, auxesis, stress, and immunity.,,
At present, the causes and pathogenesis of KD are unknown. Most scholars believe that KD is caused by one or more known or unknown pathogens that invade children with susceptibility genes, activating inflammation and other immune responses of the body, triggering cytokine release, and resulting in systemic vascular endothelial injury and dysfunction. Some scholars believe that the role of vasculitis in acute KD is as follows. Pro-inflammatory cytokines, including tumor necrosis factor-α, interleukin (IL)-6, IL-1, IL-17, IL-10, and interferon-γ, released from peripheral blood cells (e.g., monocytes, B lymphocytes, and T lymphocytes) may have direct, devastating effects on the endothelium. In recent years, several studies have indicated that Cav-1 not only regulates the activity of signal transduction pathways initiated by inflammatory cells and the inflammatory reaction but also directly regulates the release of inflammatory chemicals, including superoxide radicals, and regulates the inflammatory response. According to Garrean and other researchers, the inflammatory response of Cav-1 KO mice is lower than that of wild type mice. This implies that Cav-1 participates in and regulates the occurrence and development of the inflammatory response. Some studies have shown that Cav-1 plays a critical role in the occurrence and development of acute or chronic inflammatory reactive vascular diseases such as acute coronary syndrome, vascular restenosis, and diabetic peripheral angiopathy. However, it is unknown whether Cav-1 participates in the development of KD, and whether it affects the probability of CALs in patients with KD.
The current study found that the plasma level of Cav-1 in the acute phase group was significantly higher than that in the fever and normal control groups. This suggests that increased expression of Cav-1 promotes pathological processes associated with KD. Our other comparisons showed that the level of Cav-1 in the KD with CALs group was higher than that in the KD without CALs group. In addition, in the CALs group, the degree of coronary dilatation was positively correlated with the level of Cav-1. This indicates that Cav-1 is related to the pathogenesis of CALs. According to the ROC curve analysis, Cav-1 has high accuracy for the diagnosis of KD with CALs (AUC = 0.702, 95% CI 0.565–0.839, P = 0.007). At a Cav-1 plasma level of 58.14 ng/ml, the Youden index reached its peak value, and the sensitivity and specificity were 91.1% and 47.8%, respectively. A level of Cav-1 ≥ 58.14 ng/ml is therefore indicated for the diagnosis of KD with CALs. We speculate that there is high-level expression of Cav-1 in the acute phase of KD, that it participates in the progression and development of CALs, and that it is positively correlated with the degree of coronary dilatation. The pathogenesis behind this may be related to the stimulated and injured endothelial cells and smooth muscle cells of children with KD which can recruit inflammatory cells to the injury site through a cascade of signals. By regulating various cell signaling transduction pathways and directly regulating inflammatory factors secreted by inflammatory cells, the increasing level of Cav-1 in the blood continually promotes and aggravates the inflammatory reaction, eventually damaging the barrier of the endangium and causing coronary lesions to become aggravated. Other studies found that Cav-1, which induces vascular endothelial growth factor during angiogenesis, participates in angiogenesis. We speculate that it participates in the proliferation and repair of vascular endothelial cells during the development of the disease so that when children suffer from severe CALs, the plasma level of Cav-1 compensatorily increases for the restoration of the coronary arteries.
At present, the most effective method for controlling CALs in patients with KD is early diagnosis and high-dose globulin treatment. The pathogenesis of the IVIG effect remains unclear and possibly involves the following. First, IVIG could reduce the production of inflammatory cytokines and neutralize superantigen toxins. The pathogenesis would be relevant to anti-inflammatory factors such as autoantibodies and anti-superantigen toxin antibodies. Second, it regulates the T helper (Th) cell balance (Th1/Th2). Third, IVIG combines its Fc fragment with the target cell Fc receptor and provides negative feedback control for the overactive immune response. In our study, we found that children treated with IVIG showed a remarkable reduction in the level of Cav-1 compared to the normal group, which showed no statistically significant difference (P > 0.05). However, compared to those who had received prior treatment, there were obvious, significant differences (P < 0.05). Therefore, we speculate that the pharmacological effects of IVIG controlling CALs in KD patients could be associated with inhibiting the expression and release of Cav-1 located in the endothelium and smooth muscle cells.
| Conclusion|| |
High-level expression of Cav-1 during the acute period of KD could participate in the pathogenesis of CALs and be positively correlated with the degree of coronary dilatation. According to our ROC curve analysis, we speculate that a Cav-1 level ≥ 58.14 ng/ml is needed for a diagnosis of KD with CALs; we consider that this is a predictor of KD with CALs.
The present study has some limitations. For instance, the case number was relatively small, the follow-up period was short, and the ROC curve analysis indicated that the specificity was low (47.8%). To improve the predictive value of such an important and novel biomarker, additional multicenter studies involving a larger sample size and long-term follow-up are necessary.
We would like to thank the Children's Heart Center of the Second Affiliated Hospital and Yuying Children's Hospital for providing the research samples, and we also thank all patients who were involved in this study.
Financial support and sponsorship
This study was funded by the Medical and Health Project of Zhejiang Province (Grant no. 2017KY465, Zhejiang, China) and the Chinese Medicine Scientific Research Foundation Project of Zhejiang Province (Grant no. 2010ZA092).
Feng Zhu and Jing Huang contributed equally to this study.
Conflicts of interest
There are no conflicts of interest.
| References|| |
Newburger JW, Takahashi M, Gerber MA, Gewitz MH, Tani LY, Burns JC, et al
. Diagnosis, treatment, and long-term management of Kawasaki disease: a statement for health professionals from the committee on rheumatic fever, endocarditis, and Kawasaki disease, council on cardiovascular disease in the young, American heart association. Pediatrics 2004;114:1708-33.
Falcini F, Capannini S, Rigante D. Kawasaki syndrome: An intriguing disease with numerous unsolved dilemmas. Pediatr Rheumatol Online J 2011;9:17.
Shulman ST. IVGG therapy in Kawasaki disease: Mechanism(s) of action. Clin Immunol Immunopathol 1989;53:S141-6.
Hu G, Ye RD, Dinauer MC, Malik AB, Minshall RD. Neutrophil caveolin-1 expression contributes to mechanism of lung inflammation and injury. Am J Physiol Lung Cell Mol Physiol 2008;294:L178-86.
Voghel G, Thorin-Trescases N, Farhat N, Nguyen A, Villeneuve L, Mamarbachi AM, et al
. Cellular senescence in endothelial cells from atherosclerotic patients is accelerated by oxidative stress associated with cardiovascular risk factors. Mech Ageing Dev 2007;128:662-71.
Sonveaux P, Martinive P, DeWever J, Batova Z, Daneau G, Pelat M, et al
. Caveolin-1 expression is critical for vascular endothelial growth factor-induced ischemic hindlimb collateralization and nitric oxide-mediated angiogenesis. Circ Res 2004;95:154-61.
Beardsley A, Fang K, Mertz H, Castranova V, Friend S, Liu J. Loss of caveolin-1 polarity impedes endothelial cell polarization and directional movement. J Biol Chem 2005;280:3541-7.
Braun-Dullaeus RC, Mann MJ, Dzau VJ. Cell cycle progression: New therapeutic target for vascular proliferative disease. Circulation 1998;98:82-9.
Tsujii N, Tsuda E, Kanzaki S, Ishizuka J, Nakashima K, Kurosaki K. Late wall thickening and calcification in patients after Kawasaki disease. J Pediatr 2017;181:167-7100.
Rothberg KG, Heuser JE, Donzell WC, Ying YS, Glenney JR, Anderson RGW. Caveolin, a protein component of caveolae membrane coats. Cell 1992;68:673-82.
Zhao YY, Zhao YD, Mirza MK, Huang JH, Potula HH, Vogel SM, et al
. Persistent eNOS activation secondary to caveolin-1 deficiency induces pulmonary hypertension in mice and humans through PKG nitration. J Clin Invest 2009;119:2009-18.
Chen SF, Liou JY, Huang TY, Lin YS, Yeh AL, Tam K, et al
. Caveolin-1 facilitates cyclooxygenase-2 protein degradation. J Cell Biochem 2010;109:356-62.
Pang Q, Zhang H, Chen Z, Wu Y, Bai M, Liu Y, et al
. Role of caveolin-1/vascular endothelial growth factor pathway in basic fibroblast growth factor-induced angiogenesis and neurogenesis after treadmill training following focal cerebral ischemia in rats. Brain Res 2017;1663:9-19.
Salem AF, Bonuccelli G, Bevilacqua G, Arafat H, Pestell RG, Sotgia F, et al
. Caveolin-1 promotes pancreatic cancer cell differentiation and restores membranous E-cadherin via suppression of the epithelial-mesenchymal transition. Cell Cycle 2011;10:3692-700.
Gadjeva M, Paradis-Bleau C, Priebe GP, Fichorova R, Pier GB. Caveolin-1 modifies the immunity to Pseudomonas aeruginosa
. J Immunol 2010;184:296-302.
Yeung RS. Kawasaki disease: Update on pathogenesis. Curr Opin Rheumatol 2010;22:551-60.
Galeotti C, Kaveri SV, Cimaz R, Koné-Paut I, Bayry J. Predisposing factors, pathogenesis and therapeutic intervention of Kawasaki disease. Drug Discov Today 2016;21:1850-7.
Qin L, Zhu N, Ao BX, Liu C, Shi YN, Du K, et al
. Caveolae and caveolin-1 integrate reverse cholesterol transport and inflammation in atherosclerosis. Int J Mol Sci 2016;17:429.
Garrean S, Gao XP, Brovkovych V, Shimizu J, Zhao YY, Vogel SM, et al
. Caveolin-1 regulates NF-kappaB activation and lung inflammatory response to sepsis induced by lipopolysaccharide. J Immunol 2006;177:4853-60.
Frank PG, Hassan GS, Rodriguez-Feo JA, Lisanti MP. Caveolae and caveolin-1: Novel potential targets for the treatment of cardiovascular disease. Curr Pharm Des 2007;13:1761-9.
Burns JC, Franco A. The immunomodulatory effects of intravenous immunoglobulin therapy in Kawasaki disease. Expert Rev Clin Immunol 2015;11:819-25.
[Table 1], [Table 2], [Table 3]