Effect of NLRP3-CAMKⅡ-IRE-1α pathway induced oxidative stress on ventricular remodeling in diabetic rats

doi:10.11958/20230012

Abstract

Abstract:

Objective To explore the role and mechanism of nucleotide-binding domain-like receptor protein 3 (NLRP3) inflammasome in promoting oxidative stress enhancement through triggering calmodulin-dependent protein kinase Ⅱ (CaMKⅡ) to activate inositol demand kinase-1α (IRE-1α) in ventricular remodeling of diabetic rats. Methods Thirty-six healthy male Sprague-Dawley rats were randomly divided into the control group (CTL group), the diabetic group (DM group) and the diabetic + glibenclamide group (GLB group), with 12 rats in each group. Rats were injected 55 mg/kg streptozotocin intraperitoneally to prepare diabetic model. The GLB group was given 1.25 mg/kg GLB, an NLRP3 inhibitor, by gavage for 8 weeks since the successful modeling day. The CTL group received no intervention. After 8 weeks, blood glucose, blood pressure, body mass and ventricular body mass ratio of rats were recorded, and hemodynamic indexes were also measured. Pulmonary artery blood flow acceleration time, mean pulmonary artery pressure, systolic and diastolic ventricular septum, anterior and posterior ventricular wall thickness were evaluated by echocardiography. Epicardial activation mapping was used to measure epicardial conduction velocity, absolute heterogeneity and heterogeneity index. Western blot assay was used to detect NLRP3, caspase-1, CaMK Ⅱ, IRE-1α, niacinamide adenine dinucleotide phosphate (NOX) 2 and NOX4 protein levels. The content of reactive oxygen species (ROS) was determined by fluorescence staining. The morphology and fibrosis of ventricular tissue were observed by HE and Masson staining. Results Compared with the CTL group, blood glucose, ventricular body mass ratio, systolic and diastolic septal thickness, left ventricular anterior wall thickness increased, body mass decreased, left and right ventricular epicardial conduction velocity slowed down, and right ventricular inhomogeneity index increased in the DM group. NLRP3, caspase-1, CaMK Ⅱ, IRE-1α, NOX2 and NOX4 protein expression levels were increased, and ROS production in ventricular muscle and CVF were increased (P<0.05). Myocardial cell arrangement was disordered, and fibrosis was more obvious in the DM group. Compared with the DM group, the thickness of ventricular septum and anterior wall of left ventricle in systolic and diastolic periods were reduced in the GLB group. Compared with the DM group, epicardial conduction velocity of left and right ventricles was increased, and absolute inhomogeneity and inhomogeneity index of left ventricle were decreased in the GLB group. The protein expression levels of NLRP3, caspase-1, CaMKⅡ, IRE-1α, NOX2 and NOX4 were lower than those of the DM group. GLB reduced the production of ROS and CVF, and the ventricular myocardial fibrosis was ameliorated (P<0.05). Conclusion The NLRP3-CAMK Ⅱ-IRE-1α pathway is activated in diabetic rat ventricular myocytes to promote oxidative stress and participates in ventricular remodeling, and GLB can improve this change.

Key words: diabetes mellitus, NLR family, pyrin domain-containing 3 protein, ventricular remodeling, oxidative stress, endoplasmic reticulum stress, IRE-1α

CLC Number:

R541.75，R587.1

Figures/Tables 7

References 29

[1]	ZHANG M, SUI W, XING Y, et al. Angiotensin Ⅳ attenuates diabetic cardiomyopathy via suppressing FoxO1-induced excessive autophagy,apoptosis and fibrosis[J]. Theranostics, 2021, 11(18):8624-8639. doi:10.7150/thno.48561.
[2]	MURTAZA G, VIRK H U H, KHALID M, et al. Diabetic cardiomyopathy - A comprehensive updated review[J]. Prog Cardiovasc Dis, 2019, 62(4):315-326. doi:10.1016/j.pcad.2019.03.003.
[3]	ZHANG X, DONG S, JIA Q, et al. The microRNA in ventricular remodeling: the miR-30 family[J]. Biosci Rep, 2019, 39(8):BSR20190788. doi:10.1042/BSR20190788.
[4]	RITCHIE R H, ABEL E D. Basic mechanisms of diabetic heart disease[J]. Circ Res, 2020, 126(11):1501-1525. doi:10.1161/CIRCRESAHA.120.315913.
[5]	HE Y, CHANG Y, PENG Y, et al. Glibenclamide directly prevents neuroinflammation by targeting SUR1-TRPM4-mediated NLRP3 inflammasome activation in microglia[J]. Mol Neurobiol, 2022, 59(10):6590-6607. doi:10.1007/s12035-022-02998-x.
[6]	YANG J, YANG J, HUANG X, et al. Glibenclamide alleviates LPS-induced acute lung injury through NLRP3 inflammasome signaling pathway[J]. Mediators Inflamm, 2022, 2022:8457010. doi:10.1155/2022/8457010.
[7]	ZHAN X, CHENG L, HUO N, et al. Sodium-glucose cotransporter-2 inhibitor alleviated atrial remodeling in STZ-induced diabetic rats by targeting TLR4 pathway[J]. Front Cardiovasc Med, 2022, 9:908037. doi:10.3389/fcvm.2022.908037.
[8]	VAYKSHNORAYTE M A, OVECHKIN A O, AZAROV J E. The effect of diabetes mellitus on the ventricular epicardial activation and repolarization in mice[J]. Physiol Res, 2012, 61(4):363-370. doi:10.33549/physiolres.932245.
[9]	TIAN J H, WU Q, HE Y X, et al. Zonisamide,an antiepileptic drug,alleviates diabetic cardiomyopathy by inhibiting endoplasmic reticulum stress[J]. Acta Pharmacol Sin, 2021, 42(3):393-403. doi:10.1038/s41401-020-0461-z.
[10]	PASSARELLI M, MACHADO U F F. AGEs-induced and endoplasmic reticulum stress/inflammation-mediated regulation of GLUT4 expression and atherogenesis in diabetesmellitus[J]. Cells, 2021, 11(1):104. doi:10.3390/cells11010104.
[11]	MANGAN M S J, OLHAVA E J, ROUSH W R, et al. Targeting the NLRP3 inflammasome in inflammatory diseases[J]. Nat Rev Drug Discov, 2018, 17(9):688. doi:10.1038/nrd.2018.149.
[12]	HUANG Y, XU W, ZHOU R. NLRP3 inflammasome activation and cell death[J]. Cell Mol Immunol, 2021, 18(9):2114-2127. doi:10.1038/s41423-021-00740-6.
[13]	WU C, LU W, ZHANG Y, et al. Inflammasome activation triggers blood clotting and host death through pyroptosis[J]. Immunity, 2019, 50(6):1401-1411.e4. doi:10.1016/j.immuni.2019.04.003.
[14]	WU X, LIU Y, TU D, et al. Role of NLRP3-inflammasome/caspase-1/galectin-3 pathway on atrial remodeling in diabetic rabbits[J]. J Cardiovasc Transl Res, 2020, 13(5):731-740. doi:10.1007/s12265-020-09965-8.
[15]	QIU X, WANG Q, HOU L, et al. Inhibition of NLRP3 inflammasome by glibenclamide attenuated dopaminergic neurodegeneration and motor deficits in paraquat and maneb-induced mouse Parkinson's disease model[J]. Toxicol Lett, 2021, 349:1-11. doi:10.1016/j.toxlet.2021.05.008.
[16]	SINGH S, SHAIMA A, AHMAD S, et al. Convergence of fructose-induced NLRP3 activation with oxidative stress and ER stress leading to hepatic steatosis[J]. Inflammation, 2023, 46(1):217-233. doi:10.1007/s10753-022-01727-9.
[17]	DWIVEDI D K, JENA G B. NLRP3 inhibitor glibenclamide attenuates high-fat diet and streptozotocin-induced non-alcoholic fatty liver disease in rat: studies on oxidative stress,inflammation,DNA damage and insulin signalling pathway[J]. Naunyn Schmiedebergs Arch Pharmacol, 2020, 393(4):705-716. doi:10.1007/s00210-019-01773-5.
[18]	HEIJMAN J, MUNA A P, VELEVA T, et al. Atrial myocyte NLRP3/CaMKII nexus forms a substrate for postoperative atrial fibrillation[J]. Circ Res, 2020, 127(8):1036-1055. doi:10.1161/CIRCRESAHA.120.316710.
[19]	NIE J, TA N, LIU L, et al. Activation of CaMKII via ER-stress mediates coxsackievirus B3-induced cardiomyocyte apoptosis[J]. Cell Biol Int, 2020, 44(2):488-498. doi:10.1002/cbin.11249.
[20]	MARCINIAK S J, CHAMBERS J E, RON D. Pharmacological targeting of endoplasmic reticulum stress in disease[J]. Nat Rev Drug Discov, 2022, 21(2):115-140. doi:10.1038/s41573-021-00320-3.
[21]	REN J, BI Y, SOWERS J R, et al. Endoplasmic reticulum stress and unfolded protein response in cardiovascular diseases[J]. Nat Rev Cardiol, 2021, 18(7):499-521. doi:10.1038/s41569-021-00511-w.
[22]	HETZ C, ZHANG K, KAUFMAN R J. Mechanisms,regulation and functions of the unfolded protein response[J]. Nat Rev Mol Cell Biol, 2020, 21(8):421-438. doi:10.1038/s41580-020-0250-z.
[23]	WADGAONKAR P, CHEN F. Connections between endoplasmic reticulum stress-associated unfolded protein response,mitochondria,and autophagy in arsenic-induced carcinogenesis[J]. Semin Cancer Biol, 2021, 76:258-266. doi:10.1016/j.semcancer.2021.04.004.
[24]	YARIBEYGI H, LHAF F, SATHYAPALAN T, et al. Effects of novel antidiabetes agents on apoptotic processes in diabetes and malignancy:Implications for lowering tissue damage[J]. Life Sci, 2019, 231:116538. doi:10.1016/j.lfs.2019.06.013.
[25]	YARIBEYGI H, SATHYAPALAN T, ATKIN S L, et al. Molecular mechanisms linking oxidative stress and diabetes mellitus[J]. Oxid Med Cell Longev, 2020, 2020:8609213. doi:10.1155/2020/8609213.
[26]	IGHODARO O M. Molecular pathways associated with oxidative stress in diabetes mellitus[J]. Biomed Pharmacother, 2018, 108:656-662. doi:10.1016/j.biopha.2018.09.058.
[27]	LIU X, HUSSAIN R, MEHMOOD K, et al. Mitochondrial-endoplasmic reticulum communication-mediated oxidative stress and autophagy[J]. Biomed Res Int, 2022, 2022:6459585. doi:10.1155/2022/6459585.
[28]	POZNYAK A, GRECHKO A V, POGGIO P, et al. The diabetes mellitus-atherosclerosis connection:The role of lipid and glucose metabolism and chronic inflammation[J]. Int J Mol Sci, 2020, 21(5):1835. doi:10.3390/ijms21051835.
[29]	ZEESHAN H M, LEE G H, KIM H R, et al. Endoplasmic reticulum stress and associated ROS[J]. Int J Mol Sci, 2016, 17(3):327. doi:10.3390/ijms17030327.

组别		血糖（mmol/L）	体质量（g）		心室体质量比（‰）		收缩压（mmHg）		舒张压（mmHg）		心率（次/min）		肺动脉血流加速时间（ms）
CTL组		7.67±1.32	424.20±33.97		2.93±0.17		119.30±9.37		85.67±7.17		346.50±30.35		32.22±3.16
DM组		27.72±5.73^a	233.20±65.85^a		3.35±0.25^a		126.20±7.94		82.33±9.42		295.20±43.80		29.90±6.87
GLB组		28.33±2.24^a	267.50±68.76^a		3.08±0.12		120.50±10.77		88.17±8.35		346.80±29.71		32.58±4.72
F		62.870^＊＊	18.260^＊＊		7.830^＊＊		0.902		0.735		4.275^＊		0.476
组别	平均肺动脉压（mmHg）			收缩期室间隔厚度（mm）		舒张期室间隔厚度（mm）		收缩期左心室前壁厚度（mm）		舒张期左心室前壁厚度（mm）		收缩期左心室后壁厚度（mm）		舒张期左心室后壁厚度（mm）
CTL组	70.03±1.96			2.69±0.25		1.63±0.24		2.89±0.19		1.76±0.06		2.73±0.42		1.96±0.25
DM组	71.46±4.26			3.41±0.33^a		2.06±0.26^a		3.48±0.42^a		2.39±0.36^a		3.00±0.52		2.11±0.45
GLB组	69.80±2.92			2.62±0.26^b		1.54±0.22^b		2.24±0.32^ab		1.47±0.17^b		2.64±0.41		1.71±0.39
F	0.476			14.350^＊＊		8.134^＊＊		22.140^＊＊		24.910^＊＊		1.039		1.785

组别		血糖（mmol/L）	体质量（g）		心室体质量比（‰）		收缩压（mmHg）		舒张压（mmHg）		心率（次/min）		肺动脉血流加速时间（ms）
CTL组		7.67±1.32	424.20±33.97		2.93±0.17		119.30±9.37		85.67±7.17		346.50±30.35		32.22±3.16
DM组		27.72±5.73^a	233.20±65.85^a		3.35±0.25^a		126.20±7.94		82.33±9.42		295.20±43.80		29.90±6.87
GLB组		28.33±2.24^a	267.50±68.76^a		3.08±0.12		120.50±10.77		88.17±8.35		346.80±29.71		32.58±4.72
F		62.870^＊＊	18.260^＊＊		7.830^＊＊		0.902		0.735		4.275^＊		0.476
组别	平均肺动脉压（mmHg）			收缩期室间隔厚度（mm）		舒张期室间隔厚度（mm）		收缩期左心室前壁厚度（mm）		舒张期左心室前壁厚度（mm）		收缩期左心室后壁厚度（mm）		舒张期左心室后壁厚度（mm）
CTL组	70.03±1.96			2.69±0.25		1.63±0.24		2.89±0.19		1.76±0.06		2.73±0.42		1.96±0.25
DM组	71.46±4.26			3.41±0.33^a		2.06±0.26^a		3.48±0.42^a		2.39±0.36^a		3.00±0.52		2.11±0.45
GLB组	69.80±2.92			2.62±0.26^b		1.54±0.22^b		2.24±0.32^ab		1.47±0.17^b		2.64±0.41		1.71±0.39
F	0.476			14.350^＊＊		8.134^＊＊		22.140^＊＊		24.910^＊＊		1.039		1.785

组别	左心室心外膜传导速度（m/s）			左心室绝对不均匀性		左心室不均匀指数
CTL组	1.05±0.27			2.25±0.23		1.66±0.19
DM组	0.52±0.24^a			4.40±2.21		2.56±0.89
GLB组	1.08±0.29^b			1.68±0.20^b		1.49±0.20^b
F	7.066^＊＊			6.230^＊		5.635^＊
组别		右心室心外膜传导速度（m/s）	右心室绝对不均匀性		右心室不均匀指数
CTL组		0.87±0.20	2.32±0.47		1.38±0.13
DM组		0.58±0.15^a	4.68±3.09		2.01±0.43^a
GLB组		1.09±0.16^b	2.39±0.17		1.65±0.35
F		11.620^＊＊	2.759		4.620^＊

组别	左心室心外膜传导速度（m/s）			左心室绝对不均匀性		左心室不均匀指数
CTL组	1.05±0.27			2.25±0.23		1.66±0.19
DM组	0.52±0.24^a			4.40±2.21		2.56±0.89
GLB组	1.08±0.29^b			1.68±0.20^b		1.49±0.20^b
F	7.066^＊＊			6.230^＊		5.635^＊
组别		右心室心外膜传导速度（m/s）	右心室绝对不均匀性		右心室不均匀指数
CTL组		0.87±0.20	2.32±0.47		1.38±0.13
DM组		0.58±0.15^a	4.68±3.09		2.01±0.43^a
GLB组		1.09±0.16^b	2.39±0.17		1.65±0.35
F		11.620^＊＊	2.759		4.620^＊

组别	NLRP3	caspase-1	CaMKⅡ
CTL组	0.80±0.27	0.41±0.19	1.17±0.16
DM组	1.26±0.33^a	0.81±0.10^a	1.50±0.28^a
GLB组	0.81±0.18^b	0.36±0.20^b	1.12±0.07^b
F	5.814^＊	12.320^＊＊	7.092^＊＊
组别	IRE-1α	NOX2	NOX4
CTL组	0.28±0.09	1.15±0.34	0.92±0.22
DM组	0.56±0.13^a	1.93±0.45^a	1.60±0.21^a
GLB组	0.34±0.10^b	1.25±0.25^b	0.92±0.21^b
F	10.960^＊＊	8.459^＊＊	20.000^＊＊