Elsevier

Life Sciences

Volume 314, 1 February 2023, 121275
Life Sciences

Piperine mitigates aortic vasculopathy in streptozotocin-diabetic rats via targeting TXNIP-NLRP3 signaling

https://doi.org/10.1016/j.lfs.2022.121275Get rights and content

Abstract

Several in vivo and in vitro studies reported a favorable effect of piperine (PIP) on vascular function. However, the potential impacts of PIP on macrovasculopathy in streptozotocin (STZ)-diabetic rats have not yet been studied.

Thirty-two Sprague Dawley rats were used (n= 8/group). STZ-administered rats (50 mg/kg once, i.p) received PIP (30 mg/kg/day, orally) or its vehicle starting from day 15 till the end of the study (10 weeks). Control groups consisted of age-matched normal rats with or without PIP treatment. Metabolic and oxidative stress parameters were biochemically determined. Aortas were histologically examined. Ex vivo aortic reactivity to phenylephrine and acetylcholine was studied. Components of the TXNIP-NLRP3 pathway were assessed using real-time PCR, ELISA, and immunohistochemistry. Two-way ANOVA was used to compare groups. Statistical significance was set at P < 0.05.

PIP treatment of diabetic rats significantly reduced levels of fasting glycemia, HbA1c, and serum AGEs, TGs, TC, and LDL-C compared to control diabetic group. PIP diminished aortic endothelial denudation and fibrous tissue proliferation compared to control STZ aortas. PIP lessened aortic contractility to phenylephrine and improved aortic relaxation to acetylcholine relative to untreated STZ group. PIP administration to diabetic rats elicited significant enhancements in GSH and SOD levels, eNOS expression, and total nitrate/nitrite bioavailability compared to untreated STZ rats. Moreover, PIP attenuated aortic contents of ROS, MDA, TXNIP protein and mRNA, NF-κB p65 mRNA, NLRP3 mRNA, IL-1β protein, and caspase-3 and TNF-α expressions compared to untreated STZ levels.

In conclusion, PIP might ameliorate diabetes-associated functional and structural aortic remodeling by targeting TXNIP-NLRP3 signaling.

Introduction

Diabetes is a metabolic disease characterized by sustained levels of hyperglycemia. Diabetes is expected to affect 592 million patients by 2035 [61]. Vascular complications are a primary cause of disabilities and high mortality rates in diabetic patients [66].

Diabetic hyperglycemia is a key contributor to vascular complications, as it enhances the upregulation of advanced glycation end products (AGEs) in the circulating blood and various tissues. AGEs interact with their receptors (RAGE) in endothelial cells, leading to the production of reactive oxygen species (ROS) and the activation of nuclear factor-κB (NF-κB), which induces a chronic inflammatory response via stimulating the expression of many cytokines and growth factors [53].

ROS also enhances the activation of nucleotide-binding oligomerization domain like receptor pyrin domain containing 3 (NLRP3) inflammasome, a novel regulator of inflammation [37]. NLRP3 activation triggers the release of the proinflammatory cytokine, interleukin-1β (IL-1 β) [22], [36].

Several studies have suggested a central role of thioredoxin interacting protein (TXNIP) in the pathogenesis of diabetic vascular complications [22], [32], [36]. TXNIP elicits excessive ROS production and induces early apoptotic signals in diabetic aortic endothelium [32] and glucose-incubated endothelial cells [29], [31], [32].

Piperine (PIP) is a major alkaloid component of black pepper. Chemically, its IUPAC name is (2E, 4E)-5-(benzo[d] [1,3] dioxol-5-yl)1-(piperidin-1-yl) penta-2,4-dien-1-one. PIP is a yellow crystalline solid with molecular formula C17H19NO3, a molecular weight of 285.33 g/mol, and a melting point of 130 °C [21], [62]. PIP has been extensively evaluated for its anti-inflammatory, antioxidant, anti-mutagenic, and hepatoprotective effects [59]. PIP showed antihyperglycemic activity and enhanced the glucose-lowering effect of metformin in diabetic mice [7], [8]. Moreover, PIP attenuated the proliferation of vascular smooth muscle cells (VSMCs) by suppressing the production of ROS [30]. PIP also ameliorated glucose tolerance and reactivity of aortic rings by reducing oxidative stress and inflammation in rats with metabolic syndrome [16]. However, the potential impacts of PIP on diabetes-associated macrovasculopathy via interfering with TXNIP-NLRP3 signaling have not yet been studied.

Therefore, this study was conducted to investigate the potential effects of PIP on STZ diabetes-associated aortopathy and explore possible mechanisms involved.

Section snippets

Chemicals and reagents

PIP and STZ were from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and Sigma Aldrich Chemical Co. (St. Louis, MO, USA), respectively. All other chemicals were of the finest grade available

Animal model and grouping

Male Sprague Dawley rats, aged 6–8 weeks (250 ± 20 g), were purchased from VACSERA (Cairo, Egypt). Animals were housed under standard laboratory conditions (23 ± 2 °C; 12 h light-dark cycle; free access to standard food and water). The experimental design was accepted by the Committee of Research Ethics,

Body weight, glycemic parameters, and serum AGEs in STZ-diabetic rats

Both STZ and STZ-PIP groups showed a significant decrease in body weight relative to control rats (P < 0.0001, Fig. 1A). STZ-administered rats also showed significant elevations in fasting blood glucose (Fig. 1B), HbA1C (Fig. 1C) and serum AGE (Fig. 1D) levels by 5.7-, 1.8- and 1.3-fold, respectively, compared to those of control group. PIP treatment significantly diminished fasting blood glucose (by 27.9 %, P < 0.05), HbA1C (by 25.3 %, P < 0.001), and serum AGEs (by 19.7 %, P < 0.001) in

Discussion

In the present study, the potential impacts of PIP on type 1 diabetic vascular complications in rats were investigated. Chronic treatment with PIP successfully delayed the development of vascular complications in STZ diabetic rats via exerting antioxidant, anti-inflammatory, hypolipidemic, and hypoglycemic effects. In addition, PIP treatment restored aortic reactivity, possibly by upregulating eNOS synthase expression and attenuating endothelial apoptosis. The beneficial vascular influences of

CRediT authorship contribution statement

Manar A. Nader: Supervision, conceptualization, conceived, designed research, methodology, software, writing- reviewing, editing and approved the manuscript. Fatma M. Amin: Conducted experiments, data curation, writing- original draft preparation, software and validation. George S. G. Shehatou and Rania R. Abdelaziz: Conducted experiments, software, validation, visualization, investigation.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgment

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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