|Year : 2022 | Volume
| Issue : 3 | Page : 94-100
Elevated circulating levels of matrix metalloproteinase-9 and its association with cardiovascular risk in young smokers
Dinesh Nath, Meera Shivasekar, VM Vinodhini
Department of Biochemistry, SRM Medical College Hospital and Research Centre, SRMIST, Kattankulthur, Tamil Nadu, India
|Date of Submission||26-Jul-2021|
|Date of Decision||07-Mar-2022|
|Date of Acceptance||05-Sep-2022|
|Date of Web Publication||25-Feb-2023|
Dr. Meera Shivasekar
Department of Biochemistry, SRM Medical College Hospital and Research Centre, SRMIST, Kattankulthur - 603 203, Tamil Nadu
Source of Support: None, Conflict of Interest: None
Background: Smoking causes cardiovascular risk which may alter the stability between the production and degradation of the extracellular matrix. Matrix metalloproteinase-9 (MMP-9) is a zinc-containing endopeptidase that degrades extracellular matrix and plays a vital role in tissue remodeling. As a result, elevated serum MMP-9 levels produced by smoking, particularly in younger age, raise the risk of future coronary heart disease (CHD).
Aim and Objective: Our aim is to find out the possible relationship between circulating MMP-9 and the risk of cardiovascular disease in young smokers.
Materials and Methods: The study contains three groups. Group 1 includes 60 young active smokers with diabetic CHD, Group 2 includes 60 young active smokers with CHD, and Group 3 includes 60 healthy controls as nonsmokers who were attending SRM Medical College Hospital in Tamil Nadu for cardiology and medicine OP. Enzyme-linked immune sorbent assay was used to measure serum MMP-9, high-sensitivity C-reactive protein (hsCRP), and apolipoprotein E (APO-E) levels, and enzymatic techniques were employed to quantify lipid levels.
Results: When compared to the controls, the mean serum MMP-9, hsCRP, and APO-E levels were significantly higher in both the groups. The study also shows a significant positive association between MMP-9 with hsCRP, APO-E, smoking burden, and smoking intensity.
Conclusion: The study concludes that increased MMP-9 levels, particularly in inflammatory conditions caused by smoking, are associated with an increased risk of future cardiovascular disease.
Keywords: Apolipoprotein E, cardiovascular disease, high-sensitivity C-reactive protein, matrix metalloproteinase-9, smoking
|How to cite this article:|
Nath D, Shivasekar M, Vinodhini V M. Elevated circulating levels of matrix metalloproteinase-9 and its association with cardiovascular risk in young smokers. J Med Soc 2022;36:94-100
|How to cite this URL:|
Nath D, Shivasekar M, Vinodhini V M. Elevated circulating levels of matrix metalloproteinase-9 and its association with cardiovascular risk in young smokers. J Med Soc [serial online] 2022 [cited 2023 Mar 28];36:94-100. Available from: https://www.jmedsoc.org/text.asp?2022/36/3/94/370597
| Introduction|| |
Smoking is regarded as an environmental risk factor which is associated with different genetic factors and multifactorial disorders. The mechanism by which smoking induces cardiovascular risk is still unclear to some extent. It is well established that smoking can increase cardiovascular disease which is adversely related to the duration of cigarette smoke and the number of smoking per day. Other than active smoking, passive smoking also increases cardiovascular risk by altering the serum lipid levels in young smokers. The chemical components in cigarette smoke, especially nicotine, can increase the fatty acid level by stimulating the release of adrenaline, leading to an increase in the cardiovascular risk. Free fatty acid can increase the triglyceride level (TGL) and low-density lipoprotein (LDL) secretion and release cholesterol from the hepatic circulation. Smoking also produces free radicals which can alter the coagulation system, and may accelerate the formation of plaque in the arteries. Smoking also induces inflammation which may lead atherosclerotic plaque formation and plays an important role for the development of cardiovascular disease. Most recently, different studies show that the formation of atherosclerotic plaque and subsequent rupture happens due to the development of chronic inflammation. Although for the predictor of cardiovascular disease, the markers for general inflammation are related to acute phase reactants and pro-inflammatory cytokines including high-sensitivity C-reactive protein (hsCRP). Most particularly, the tissue necrosis factor-α and pro-inflammatory cytokines such as interleukin-1 (IL-1) upregulate matrix metalloproteinase-9 (MMP-9) synthesis by mesenchymal stem cells., Recent studies reveal that smoking may alter the stability of extracellular matrix by altering the balance between the production and degradation of MMP, which may lead to the development of cardiovascular disease. Matrix metalloproteinase are the families of zinc-containing zymogene endopeptidase that degrade extracellular matrix proteins, play a pivotal role in extracellular matrix for tissue remodeling, and contribute in a multiplicity physiological process.,, Among the MMP family, MMP-9 is a 92 KDa collagenase enzyme. In the basement membrane, MMP-9 breaks proteoglycan proteins, type 4 collagen, and interstitial proteins., Till date, it is the most studied enzyme in relation to cardiovascular disease. Different studies show that, at high inflammatory condition, the MMP-9 activity has been increased in the formation of atherosclerotic plaque., Hence, the increased level of MMP-9, especially in the inflammatory condition caused by smoking, is at a higher risk of future cardiovascular disease. Thus, MMP-9 can be used as a therapeutic target as well as a biomarker for future coronary heart disease (CHD)., Hence, being a part of India in the southern state, the smoking rate in Tamil Nadu is vulnerably increased, which may increase the prevalence of CHD patients in recent times, which may be due to the modification of lifestyle and the increase rate of smoking in early age. Thus, the current study intends to investigate the link between circulating MMP-9 and the risk of CHD in young smokers.
| Materials and Methods|| |
Study design and subjects
The present cross-sectional study was conducted at SRM Medical College Hospital and Research Center, SRMIST, Tamil Nadu, India, between October 2019 and September 2021. The study group was divided into three groups. Group 1 includes 60 young active smokers with diabetic CHD, Group 2 includes 60 young active smokers with CHD, and Group 3 includes 60 healthy controls as nonsmokers who were attending SRM Medical College Hospital in Tamil Nadu for cardiology and medicine OP. A standard questionnaire is used during regular cardiovascular health assessments to acquire information about the patient's history and lifestyle characteristics. Patients who smoked on a regular basis and had been diagnosed with CHD met the inclusion criteria. The diagnosis of CHD was based on abnormal coronary angiography with more than 50% stenosis in one or more major arteries. The study excludes the participants with cardiomyopathy, chronic diseases such as liver failure, cancer patients, heart failure, pregnancy, cardiovascular accidents, significant systemic sickness, and systemic inflammatory disease.
Ascertainment of smoking exposure
The Smoking habits were ascertained through self-reporting. Males aged between (20-55 years) who were either non-smoker or at least 5 cigarettes per day for more than one-year duration of smoking.
Ascertainment of covariates
Self-reporting is used to determine sociodemographic factors such as age, gender, educational history, and other health and medical histories. Standard equipment and technique were used to measure anthropometric characteristics such as weight and height. After 2 min of rest, resting blood pressure (BP) was measured three times with 1-min intervals in the sat posture; the average of the second and third measurements was used for analysis. A systolic BP of more than 140 mmHg and a diastolic BP of more than 90 mmHg were defined as hypertension. Diabetes mellitus was defined as a previous medical diagnosis of diabetes mellitus or meeting diagnostic criteria for diagnosis based on fasting plasma glucose levels of >126 mg/dl, 2-h plasma glucose levels obtained as part of a 75-g oral glucose tolerance test >200, or the glycated hemoglobin test of >6.5%.
A sandwich enzyme-linked immunosorbent assay (ELISA) was used to determine the concentrations of serum MMP-9, hsCRP, and apolipoprotein-E (ELISA). The AU480 automated analyzer was used to assess fasting serum glucose levels, total cholesterol (TC), TGLs, serum high-density lipoprotein cholesterol (HDL-C) levels, low-density cholesterol levels, and serum very LDL levels.
Ethical approval for the study was obtained from the Human Research Ethical Committee of SRM Medical College Hospital and Research Center, SRMIST (approval number IEC No: 1763). All study participants provided informed consent.
The statistical analysis was performed using IBM Corp.'s Statistical Package for the Social Sciences (SPSS), version 22 (Armonk, NY, USA). The quantitative variables are expressed as mean and standard deviation (SD). Data were examined using one-way analysis of variance to compare the differences between the three groups. Differences were considered to be highly significant, significant, or nonsignificant for P < 0.001, P < 0.05, or P > 0.05, respectively. The associations between variables were determined using Pearson's correlation coefficient (r).
| Results|| |
Baseline and biochemical characteristics of the study groups
[Table 1] and [Table 2] show the demographic and baseline characteristic data of all the three groups. The subjects' weight, body mass index (BMI), waist-to-hip (W/H) ratio, BP, number of cigarettes per day (smoking intensity), and smoking duration all showed a significant difference (smoking burden). When compared to controls, smokers in Groups 1 and 2 had substantially higher fasting blood glucose (FBG) and lipid levels. The study group was not receiving any kind of a lipid-lowering treatment.
|Table 1: Anthropometric measurements of smokers with coronary heart disease and normal controls|
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|Table 2: Biochemical characteristics of Group 1, Group 2, and normal controls (nonsmokers)|
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The serum MMP-9, hsCRP, and apolipoprotein E (APO-E) levels were represented as mean ± SD in [Table 2]. The result from the study revealed a gradual increase in serum MMP-9, hsCRP, and APO-E levels in smokers with CHD followed by the diabetic CHD subjects when compared to controls (P < 0.0001), indicating that smokers induce inflammation.
As shown in [Table 3], the study shows Pearson's correlation between serum MMP-9, hsCRP, and APO-E in diabetic CHD subjects. We found a positive correlation between MMP-9 and hsCRP (r = 0.3776), APO-E (r = 0.4614), BMI (r = 0.4673), FBG (r = 0.3978), TC (r = 0.4858), TGL (r = 0.3917), LDL (r = 0.4689), TC/HDL (r = 0.3225), number of smoking/day (r = 0.4287), duration of smoking (r = 0.3638), and a negative correlation was found with HDL-C levels.
|Table 3: Correlation of all different parameters in smokers with diabetic coronary heart disease subjects|
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As shown in [Table 4], the study also shows Pearson's correlation between serum MMP-9, hsCRP, and APO-E in nondiabetic CHD subjects. We also found a positive correlation between MMP-9 and hsCRP (r = 0.3776), APO-E (r = 0.4039), BMI (r = 0.734), W/H ratio (r = 0.4089), FBG (r = 0.3105), TC (r = 0.3204), TGL (r = 0.3881), LDL-C (r = 0.5003), TC/HDL-C (r = 0.4071), LDL-C/HDL-C (r = 0.4926), number of smoking/day (r = 0.3411), and duration of smoking (r = 0.3175).
|Table 4: Correlation of all different parameters in smokers with nondiabetic coronary heart disease subjects|
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Along with this, we also performed a linear regression analysis to see whether the relationship between MMP-9 with hsCRP, APO-E, and smoking intensity (number of smoking per day) was affected by smoking status [Figure 1].
|Figure 1: Linear regression analysis of MMP-9 levels with other biochemical parameters in smokers with diabetic CHD subjects, MMP-9: Matrix metalloproteinase-9, CHD: Coronary heart disease. (a) MMP-9 with APO-E, (b) MMP-9 with hsCRP, (c) MMP-9 with BMI, (d) MMP-9 with HDL, (e) MMP-9 with LDL, (f) MMP-9 with smoking/day|
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| Discussion|| |
To our information, this report is the first-ever report on MMP-9 with respect to young smokers. Smoking habit continuously elevates the risk of cardiovascular disease and peripheral vascular disease. Modifiable risk factor such as high BP or high cholesterol level does not explain clearly the relation between cigarette smoke and the occurrence of cardiovascular disease risk., However, certain studies give evidence that smoking increases the circulating MMP-9 concentration, which increases the risk of cardiovascular disease.
This current study shows a significant positive correlation between the cardiovascular risk factors, especially smoking, and the inflammatory marker (hsCRP) with MMP-9 in young smokers as it is compared to nonsmokers with a significant P < 0.05. Moreover, different studies show that the elevated levels of MMP-9 were positively associated with smoking status and inflammatory markers (including hsCRP and IL-6)., The study also shows that MMP-9 is significantly correlated with APO-E levels. This may be due to the shedding of the lipoprotein receptor by MMP-9. MMP-9 is an endopeptidase that can bind and proteolysis (i.e., shedding) lipoprotein receptors.
In this study, the serum level of MMP-9 was significantly increased in smokers with CHD (with and without diabetes) than the control group (P < 0.0001). MMP-9 levels were significantly increased in Group 1 when compared to Group 2 with a significant value of (<0.001), which may reflect abnormal extracellular matrix metabolism in the diabetic CHD group., The study also identifies MMP-9 as a new modulator of cholesterol metabolism. Furthermore, the results suggest that dysregulation of MMP-9 activity can alter the hepatic transcriptional responses to dietary cholesterol, resulting in metabolic disorders that could lead to atherosclerosis and CHD.
Smoking accelerates inflammation and oxidative modification of lipids and prospectively slows down the matrix metalloproteinase activity at a various different level. By activating inflammatory transcription factors, smoking increases MMP expression. Along with this, smoking also elevates the monocyte expression of IL-beta cells. Cigarette smoke contains nicotine and predominant metabolite cotinine increases the production of vascular smooth muscle cell collagenase and gelatinase may lead to plaque rupture.
Different mechanisms have been anticipated to explain by which smoking induces the stimulation of MMP, both in vitro and in vivo., In vitro tobacco smoke induces MMP-9 expression through the endothelial cell. Similarly, exposure of smoking induces MMP-1 expression through human fibroblast. While tobacco smoke also induces proteolysis by inhaling the cadmium present in smoke, which may lead to cardiovascular disease as it increases in the aorta of smokers.
The major limitation of this study is the limited number of sample size. Other than this, we did not see the genetic polymorphisms of the complete set of data which may be able to explain in some more extent in the distribution of the variance of MMP-9 in young smokers. Nevertheless, we need further study on smoking which may reflect the MMP-9 concentration in young smokers. It is very important to analyze the activity of endogenous tissue inhibitors of metalloproteinase 1 (TIMP-1), because it creates a balance between the MMP-9 and TIMP-1.,, However, our study did not analyze the TIMP-1 concentration.
Hence, our finding on MMP-9 in young smokers gives evidence that cigarette smoking accelerates the circulating MMP-9 levels in young age.
| Conclusion|| |
The current investigation revealed a substantial link between serum MMP-9 and the risk of CHD among young smokers. According to the findings, an increase in MMP-9 levels, particularly in inflammatory conditions produced by smoking, is associated with a greater risk of future cardiovascular disease
The authors acknowledge the Department of Medicine, Cardiology and Master Health Check-up Unit for the permitting and supporting.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Table 1], [Table 2], [Table 3], [Table 4]