Abstract
Background
PCSK9 gene expression is associated with biological processes such as lipid metabolism, glucose metabolism, and inflammation. In the present study, our primary objective was to assess the association between the single-nucleotide polymorphisms in the PCSK9 gene and type 2 diabetes in Uygur subjects, in **njiang, China.
Methods
We designed a case–control study including 662 patients diagnosed with T2DM and 1220 control subjects. Four single-nucleotide polymorphisms (rs11583680, rs2483205, rs2495477 and rs562556) of PCSK9 gene were genotyped using the improved multiplex ligation detection reaction technique.
Results
For rs2483205, the distribution of genotypes, dominant model (CC vs CT + TT), overdominant model (CC + TT vs CT) showed significant differences between T2DM patients and the controls (P = 0.011 and P = 0.041 respectively). For rs2495477, the distribution of genotypes, the dominant model (AA vs GA + GG) showed significant differences between T2DM patients and the controls (P = 0.024). Logistic regression analysis suggested after adjustment of other confounders, the differences remained significant between the two groups [for rs2483205 CC vs CT + TT: odds ratio (OR) = 1.321, 95% confidence interval (CI) 1.078–1.617, P = 0.007; CC + TT vs CT: OR = 1.255, 95% CI 1.021–1.542, P = 0.03; for rs2495477 AA vs GA + GG: OR = 1.297, 95% CI 1.060–1.588, P = 0.012].
Conclusion
The present study indicated that CT + TT genotype and CT genotype of rs2483205, as well as GA + GG genotype of rs2495477 in PCSK9 gene were associated with an increased risk of type 2 diabetes in the Uygur population in **njiang.
Similar content being viewed by others
Introduction
Type 2 diabetes mellitus (T2DM) is a complex metabolic disease mainly characterized by hyperglycemia arising from insulin resistance and/or insufficient insulin secretion [1]. Long-term chronic hyperglycemia can lead to multiple system damage and failure, such as heart, eyes, kidneys, nerves, and blood vessels [2]. Survey data show that the prevalence of T2DM has been rapidly rising worldwide and is projected to grow to 440 million by 2030 [3]. T2DM has become one of the serious public health problems. T2DM is the result of the interaction between genetic and environmental factors, a large number of studies have revealed that multiple single nucleotide polymorphisms (SNPs) are related to the development of T2DM [4, 5].
Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a secreted protein that is mainly expressed in the liver, small intestinal epithelial cells, neural tissue, and kidney cells [6]. The function of PCSK9 is thought to increase the amount of low-density lipoprotein cholesterol (LDL-C) in the blood by degrading the low-density lipoprotein receptor (LDLR) that transports LDL-C [7]. In recent years, PCSK9 is one of the promising hot targets in the field of cardiovascular research [8], and the PCSK9 antibody has been used to treat some patients with familial hypercholesterolemia and statin tolerance [9,10,11,12]. Qiu et al. [13] found genetic polymorphisms of PCSK9 associated with cardiovascular disease. Several studies have demonstrated that polymorphisms of PCSK9 was associated with the plasma lipid levels in Chinese [14, 15].
Interestingly, it has been found that PCSK9 is also associated with the biological processes of glucose metabolism. Saavedra et al. [16] found that the incidence of diabetes and prediabetes was twice as high in familial hypercholesterolemia (FH) individuals carrying the PCSK9-InsLEU gene mutation as in non-carriers. Mohamed et al. demonstrated that PCSK9 relative expression levels and the E670G (rs505151) AG genotype are cardiovascular disease risk factors among Egyptian diabetics [17]. However, the relationship between PCSK9 genetic polymorphisms and T2DM remains unclear. Thus, the current case–control study was designed to explore the possible correlation of PCSK9 gene polymorphisms with T2DM among Uygur Chinese populations in **
Blood samples were collected from all participants, and genomic DNA was extracted from peripheral blood leukocytes using a whole blood genome extraction kit (Bei**g Bioteke Corporation, Bei**g, China). We obtained four tag SNPs of PCSK9 (rs11583680, rs2483205, rs2495477, and rs562556) according to Haploview 4.0 software and the International HapMap Project website (http://hapmap.ncbi.nlm.nih.gov/index.html.en) by using minor allele frequency (MAF) ≥ 0.05 and linkage disequilibrium patterns with r2 ≥ 0.8 as a cut-off. The four SNPs of the PCSK9 gene were genotyped by improved multiple ligase detection reaction (iMLDR) genoty** assays.
Laboratory examination
Peripheral venous blood samples (5 mL) were collected in EDTA-containing tubes from all participants following overnight fasting greater than 8 h for biochemical assays. Biochemical variables, including serum concentrations of creatinine (Cr), blood urea nitrogen (BUN), uric acid (UA), fasting blood glucose (FBG), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C), were measured by using standard methods in the Department of Clinical Laboratory at the First Affiliated Hospital of ** type 2 diabetes mellitus. JAMA. 2001;286(3):327–34." href="#ref-CR32" id="ref-link-section-d8423468e4462">32,33,34]. While PCSK9 can induce a variety of cells and tissues to secretes pro-inflammatory factors, it is therefore speculated that PCSK9 may be involved in the pathogenesis of T2DM by promoting an inflammatory response [35, 36]. Some studies are supporting those loss-of-function genetic variants of the PCSK9 gene are associated with increased risk of T2DM [37]. It may be because PCSK9 deficiency can lead to increased expression of LDLR in pancreatic β-cells which leads to accumulation of cholesteryl esters, inhibition of islet function and insulin secretion, and causes elevated blood glucose [38].
In the present study, we genotyped polymorphisms of four SNPs in the PCSK9 gene and found that rs2483205 and rs2495477 were associated with T2DM. Rs2495477 is located in the fifth intron of the PCSK9 gene and affects the splicing process of RNA, which may lead to reduced PCSK9 mRNA levels [39]. Rs2483205 is also located in the intronic region of the PCSK9 gene and regulates the promoter flanking regions. It has been shown that rs2483205 is associated with decreased LDL cholesterol concentrations [40]. The CC genotype and CC + TT genotype of rs2483205, the rs2495477 AA genotype were very common in the T2DM patients compared with the control subjects in the Uygur population in **njiang. After adjusting for confounders, the rs248320 CC, the rs2483205 CC + TT, and the rs2495477 AA were still independent risk factors for T2DM. We hypothesize that rs2483205 and rs2495477 polymorphisms may be involved in the development of diabetes by decreasing plasma PCSK9 levels, leading to impaired islet function.
There are some limitations to the study. Firstly, our study population was limited to the First Affiliated Hospital of **njiang Medical University and may have suffering some risk factors of T2DM. Secondly, as this was an observational study, we cannot definitively establish cause and effect. Thirdly, the present study lacked functional validation of studied SNPs, additional studies need to be conducted to demonstrate the molecular mechanism between PCSK9 gene polymorphisms and T2DM.
Conclusions
In conclusion, our study suggests that genetic polymorphisms in the PCSK9 gene are associated with T2DM in Uygur subjects in **njiang. Subjects with CC genotype or CC + TT genotype of rs2483205 as well as subjects with AA genotype of rs2495477 were associated with an increased risk of T2DM. However, the mechanisms that may link PCSK9 gene polymorphisms to T2DM remain unclear.
OR, odds ration; CI, confidence interval
Availability data and materials
All data generated or analyzed during this study are included in this published article.
Abbreviations
- T2DM:
-
Type 2 diabetes mellitus
- PCSK9:
-
Proprotein convertase subtilisin/kexin type 9
- SNPs:
-
Single nucleotide polymorphisms
- SBP:
-
Systolic blood pressure
- DBP:
-
Diatolic blood pressure
- UA:
-
Uric acid
- FBG:
-
Fasting blood glucose
- NGT:
-
Normal glucose tolerance
- iMLDR:
-
Improved multiple ligase detection reaction
- SREBP-1c:
-
Sterol regulatory element-binding protein-1c
- BMI:
-
Body mass index
- Cr:
-
Creatinine
- BUN:
-
Blood urea nitrogen
- GLU:
-
Glucose
- TG:
-
Triglyceride
- TC:
-
Total cholesterol
- HDL-C:
-
High-density lipoprotein cholesterol
- LDL-C:
-
Low-density lipoprotein cholesterol
- LDLR:
-
Low-density lipoprotein receptor
References
Montemurro C, Nomoto H, Pei L, et al. IAPP toxicity activates HIF1α/PFKFB3 signaling delaying β-cell loss at the expense of β-cell function. Nat Commun. 2019;10(1):2679.
Suryavanshi SV, Barve K, Addepalli V, et al. Triphala churna-A traditional formulation in Ayurveda mitigates diabetic neuropathy in rats. Front Pharmacol. 2021;12: 662000.
Yan J, Tie G, Wang S, et al. Diabetes impairs wound healing by Dnmt1-dependent dysregulation of hematopoietic stem cells differentiation towards macrophages. Nat Commun. 2018;9(1):33.
Ye Z, Sharp SJ, Burgess S, et al. Association between circulating 25-hydroxyvitamin D and incident type 2 diabetes: a mendelian randomisation study. Lancet Diabetes Endocrinol. 2015;3(1):35–42.
Aboelkhair NT, Kasem HE, Abdelmoaty AA, et al. TCF7L2 gene polymorphism as a risk for type 2 diabetes mellitus and diabetic microvascular complications. Mol Biol Rep. 2021;48:5283–90.
Blanchard V, Khantalin I, Ramin-Mangata S, et al. PCSK9: from biology to clinical applications. Pathology. 2019;51(2):177–83.
Horton JD, Cohen JC, Hobbs HH. Molecular biology of PCSK9: its role in LDL metabolism. Trends Biochem Sci. 2007;32(2):71–7.
Qi Z, Hu L, Zhang J, et al. PCSK9 (Proprotein Convertase Subtilisin/Kexin 9) enhances platelet activation, thrombosis, and myocardial infarct expansion by binding to platelet CD36. Circulation. 2021;143(1):45–61.
Giugliano RP, Pedersen TR, Park JG, et al. Clinical efficacy and safety of achieving very low LDL-cholesterol concentrations with the PCSK9 inhibitor evolocumab: a prespecified secondary analysis of the FOURIER trial. Lancet. 2017;390(10106):1962–71.
Sabatine MS, De Ferrari GM, Giugliano RP, et al. Clinical benefit of evolocumab by severity and extent of coronary artery disease: analysis from FOURIER. Circulation. 2018;138(8):756–66.
White HD, Steg PG, Szarek M, et al. Effects of alirocumab on types of myocardial infarction: insights from the ODYSSEY OUTCOMES trial. Eur Heart J. 2019;40(33):2801–9.
Razek O, Cermakova L, Armani H, et al. Attainment of recommended lipid targets in patients with familial hypercholesterolemia: real-world experience with PCSK9 inhibitors. Can J Cardiol. 2018;34(8):1004–9.
Qiu C, Zeng P, Li X, et al. What is the impact of PCSK9 rs505151 and rs11591147 polymorphisms on serum lipids level and cardiovascular risk: a meta-analysis. Lipids Health Dis. 2017;16(1):111.
Li Z, Zhao T, Tan X, et al. Polymorphisms in PCSK9, LDLR, BCMO1, SLC12A3, and KCNJ1 are Associated with Serum Lipid Profile in Chinese Han Population. Int J Environ Res Public Health.2019;16(17).
Chuan J, Qian Z, Zhang Y, et al. The association of the PCSK9 rs562556 polymorphism with serum lipids level: a meta-analysis. Lipids Health Dis. 2019;18(1):105.
Saavedra YGL, Dufour R, Baass A. Familial hypercholesterolemia: PCSK9 InsLEU genetic variant and prediabetes/diabetes risk. J Clin Lipidol. 2015;9(6):786–93.
Mohamed SH, Hassaan MMM, Ibrahim BA, et al. PCSK9 E670G (rs505151) variant and coronary artery disease risk among diabetics. Genet Test Mol Biomark. 2021;25(9):615–23.
American Diabetes Association Professional Practice Committee. 2. Classification and Diagnosis of Diabetes: Standards of Medical Care in Diabetes-2022. Diabetes Care.2022 01 01 ;45(Suppl 1): S17-S38.
Aronow WS, Fleg JL, Pepine CJ, et al. ACCF/AHA 2011 expert consensus document on hypertension in the elderly: a report of the American College of Cardiology Foundation Task Force on Clinical Expert Consensus documents developed in collaboration with the American Academy of Neurology, American Geriatrics Society, American Society for Preventive Cardiology, American Society of Hypertension, American Society of Nephrology, Association of Black Cardiologists, and European Society of Hypertension. J Am Coll Cardiol. 2011;57(20):2037–114.
Malo J, Parajuli A, Walker SW. PCSK9: from molecular biology to clinical applications. Ann Clin Biochem. 2020;57(1):7–25.
Li HH, Li J, Zhang XJ, et al. 23,24-Dihydrocucurbitacin B promotes lipid clearance by dual transcriptional regulation of LDLR and PCSK9. Acta Pharmacol Sin. 2020;41(3):327–35.
Urban D, Pöss J, Böhm M, et al. Targeting the proprotein convertase subtilisin/kexin type 9 for the treatment of dyslipidemia and atherosclerosis. J Am Coll Cardiol. 2013;62(16):1401–8.
Norata GD, Tavori H, Pirillo A, et al. Biology of proprotein convertase subtilisin kexin 9: beyond low-density lipoprotein cholesterol lowering. Cardiovasc Res. 2016;112(1):429–429.
Tromp TR, Stroes ESG, Hovingh GK, et al. Gene-based therapy in lipid management: the winding road from promise to practice. Expert Opin Investig Drugs. 2020;29(5):483–93.
Pott J, Schlegel V, Teren A, et al. Genetic regulation of PCSK9 (proprotein convertase subtilisin/kexin type 9) plasma levels and its impact on atherosclerotic vascular disease phenotypes. Circ Genom Precis Med. 2018;11(5): e001992.
Schuster S, Rubil S, Endres M, et al. Anti-PCSK9 antibodies inhibit pro-atherogenic mechanisms in APOE*3Leiden.CETP mice. Sci Rep.2019 07 31 ;9(1):11079.
Shi J, Zhang W, Niu Y, et al. Association of circulating proprotein convertase subtilisin/kexin type 9 levels and the risk of incident type 2 diabetes in subjects with prediabetes: a population-based cohort study. Cardiovasc Diabetol. 2020;19(1):209.
Costet P, Cariou B, Lambert G, et al. Hepatic PCSK9 expression is regulated by nutritional status via insulin and sterol regulatory element-binding protein 1c. J Biol Chem. 2006;281(10):6211–8.
Miao J, Manthena PV, Haas ME, et al. Role of insulin in the regulation of proprotein convertase subtilisin/kexin type 9. Arterioscler Thromb Vasc Biol. 2015;35(7):1589–96.
Momtazi AA, Banach M, Pirro M, et al. PCSK9 and diabetes: is there a link? Drug Discov Today. 2017;22(6):883–95.
Awan Z, Dubuc G, Faraj M, et al. The effect of insulin on circulating PCSK9 in postmenopausal obese women. Clin Biochem. 2014;47(12):1033–9.
Pradhan AD, Manson JE, Rifai N, et al. C-reactive protein, interleukin 6, and risk of develo** type 2 diabetes mellitus. JAMA. 2001;286(3):327–34.
Donath MY. Targeting inflammation in the treatment of type 2 diabetes: time to start. Nat Rev Drug Discov. 2014;13(6):465–76.
Goldfine AB, Shoelson SE. Therapeutic approaches targeting inflammation for diabetes and associated cardiovascular risk. J Clin Invest. 2017;127(1):83–93.
Liu S, Deng X, Zhang P, et al. Blood flow patterns regulate PCSK9 secretion via MyD88-mediated pro-inflammatory cytokines. Cardiovasc Res. 2020;116(10):1721–32.
Ding Z, Pothineni NVK, Goel A, et al. PCSK9 and inflammation: role of shear stress, pro-inflammatory cytokines, and LOX-1. Cardiovasc Res. 2020;116(5):908–15.
Schmidt AF, Swerdlow DI, Holmes MV, et al. PCSK9 genetic variants and risk of type 2 diabetes: a mendelian randomisation study. Lancet Diabetes Endocrinol. 2017;5(2):97–105.
Da Dalt L, Ruscica M, Bonacina F, et al. PCSK9 deficiency reduces insulin secretion and promotes glucose intolerance: the role of the low-density lipoprotein receptor. Eur Heart J. 2019;40(4):357–68.
Pott J, Gådin JR, Theusch E, et al. Meta-GWAS of PCSK9 levels detects two novel loci at APOB and TM6SF2. Hum Mol Genet. 2022;31(6):999–1011.
Gai MT, Adi D, Chen XC, et al. Polymorphisms of rs2483205 and rs562556 in the PCSK9 gene are associated with coronary artery disease and cardiovascular risk factors. Sci Rep. 2021;11(1):11450.
Acknowledgements
We are grateful to the patients involved in the study.
Funding
This work was supported financially by the National Natural Science Foundation of China (No. 81960097).
Author information
Authors and Affiliations
Contributions
MMW and CFL conceived the study, designed, and performed the experiments; YLT and GY participated in the collection of data; SQY provided direction and guidance on the whole experiment; MMW and CFL wrote the first draft of the paper, XM, YTM, and BZW critically revised the manuscript. All authors read and approved the final manuscript.
Corresponding authors
Ethics declarations
Ethical approval and consent to participants
We conducted the study in accordance with the Declaration of Helsinki. Each participant gave written informed consent and explicit permission for pertinent clinical data collection and DNA analyses. The study was approved by the Ethics Committee of the First Affiliated Hospital of **njiang Medical University in **njiang, China.
Consent for publication
Not applicable.
Competing interests
The authors declare that they have no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Additional file 1: Figure S1.
The distribution of four SNPs genotypes in the case group and control group.
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
About this article
Cite this article
Wang, MM., Lu, CF., Yan, Sq. et al. Association of genetic polymorphisms of PCSK9 with type 2 diabetes in Uygur Chinese population. BMC Cardiovasc Disord 22, 284 (2022). https://doi.org/10.1186/s12872-022-02710-w
Received:
Accepted:
Published:
DOI: https://doi.org/10.1186/s12872-022-02710-w