Carvedilol suppresses cartilage matrix destruction

https://doi.org/10.1016/j.bbrc.2016.10.032Get rights and content

Highlights

  • Carvedilol abolishes IL-1β induced degradation of col Ⅱ and aggrecan.

  • Carvedilol inhibits the expression of MMP-1 and MMP-3 induced by IL-1β.

  • Carvedilol attenuates phosphorylation of IKK-α/β and degradation of IκB-α.

  • Carvedilol inhibits nuclear translocation of p65 and NF-κB luciferase activity.

Abstract

Collagen type Ⅱ (col Ⅱ) and aggrecan, the main components of the extracellular matrix (ECM) in human joint cartilage, have been reported to be reduced by chronic production of inflammatory cytokine interleukin (IL)-1β in arthritic joints. Carvedilol, a licensed medicine, has been used for treatment of hypertension, congestive heart failure and coronary disease in clinics. In this study, we investigated the effects of Carvedilol on the expression of col Ⅱ and aggrecan. Our results demonstrate that treatment with Carvedilol didn't change the expression of aggrecan or col Ⅱ at mRNA levels in SW1353 chondrocytes. However, the expression of aggrecan and Col II at protein levels were significantly reduced by IL-1β treatment, which were reversed by Carvedilol in a dose dependent manner, suggesting the inhibitory effects of Carvedilol on the expression of aggrecan and Col II are at post-translational modification levels. In addition, it was shown that IL-1β treatment highly induced MMP-1 and MMP-13 expression in SW1353 chondrocytes at both gene and protein expression levels, which were restored by Carvedilol in a dose dependent manner. Mechanistically, exposure to IL-1β increased phosphorylation of IKK-α/β and degradation of IκB-α in SW1353 chondrocytes, which were suppressed by pretreatment with Carvedilol. Administration of Carvedilol inhibited IL-1β-induced translocation of NF-κB p65 from cytosol to nucleus manner. Notably, a luciferase reporter assay showed that IL-1β severely increased NF-κB luciferase activity, which was markedly suppressed by Carvedilol treatment. Our results suggest that Carvedilol might be a potential therapeutic agent for chondro-protective therapy.

Introduction

The extracellular matrix (ECM) in human joint cartilage consists of collagens, elastin, proteoglycans and their large aggregates [1]. Among them, collagen type Ⅱ (col Ⅱ) and aggrecan are the main components of ECM materials, which play a pivotal role in providing support and tensile strength for the joint [2]. Increasing evidence has shown that the abundant extracellular matrix of articular cartilage has to be maintained by a limited number of chondrocytes. The denaturation and degradation of ECM materials, especially col Ⅱ in the cartilage and synovial fluid leads to progressive loss of integrity and strength of the cartilage, which is one of the most important pathological characteristics in osteoarthritis (OA). Chronic production of inflammatory cytokine interleukin (IL)-1β in arthritic joints by activated synovial cells and infiltrating macrophages, is considered one of the most potent catabolic factors in arthritis [3]. Overproduction of IL-1β activates chondrocytes, which in turn produce catabolic factors such as matrix metalloproteinases (MMPs), nitric oxide (NO), and other proinflammatory cytokines [4]. MMPs are a large group of enzymes that degrade a wide variety of ECM components. At the basal levels, MMPs take responsible for the normal turn-over of ECM materials. However, under special pathological conditions, chondrocytes highly induce MMPs, which lead to the degradation of ECM components [5]. Among these, MMP-1 and -13 are of particular importance because they are elevated in joint disorders and degrades cartilage collagens [6]. Inhibiting the activity of MMPs has become a potential strategy to prevent and/or treat osteoarthritic symptoms and several other ECM-degradation diseases to some extent.

Carvedilol, a non-specific β-adrenergic blocker, has been used for treatment of hypertension, congestive heart failure and coronary disease in clinics [7]. Interestingly, several in vivo and in vitro studies demonstrated the cardio-protective [8], nephron-protective [9] and hepato-protective effects of carvedilol against various toxicants induced preclinical models. And these effects are independent of its vasodilator and β-adrenergic blocking properties. Notably, increasing evidence has shown that Carvedilol possesses distinctive antioxidant property with both ROS-scavenging and ROS-suppressive effects [10], [11]. In addition, Carvedilol also displays its anti-inflammatory properties through reducing pro-inflammatory cytokine production combined with increased anti-inflammatory cytokine (e.g., IL-10) production [12]. However, few studies have examined the therapeutic potential of Carvedilol for cartilage degradation. In this study, we aimed to investigate whether Carvedilol has a protective effect against IL-1β induced expression of MMPs and degradation of col Ⅱand aggrecan.

Section snippets

Cell culture

The human chondrosarcoma cell line SW1353 cells were obtained from American type culture collection (ATCC HTB-94, USA). Cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 60 U/mL penicillin, 60 μg/mL streptomycin, and 2 mM glutamine at 37 °C. SW1353 cells were pretreated with the vehicle (DMSO, 0.01%, v/v) or Carvedilol (10 and 50 μM) for 24 h and then stimulated with IL-1β (10 ng/mL) for 24 h.

RNA isolation and real-time PCR

Total intracellular RNA from cultured

Results

Firstly, we detected the effects of Carvedilol on the expression of aggrecan and Col II in IL-1β-stimulated SW1353 chondrocytes. SW1353 chondrocytes were pretreated with the vehicle (DMSO, 0.01%, v/v) or Carvedilol (10 and 50 μM) for 24 h and then stimulated with IL-1β (10 ng/mL) for another 24 h. Results in Fig. 1A demonstrate that neither the expression of aggrecan nor the expression of Col II at mRNA levels was significantly changed. However, the expression of aggrecan and Col II at protein

Discussion

Carvedilol, a non-selective β-adrenergic receptor blocker, has been reported to possess multi-functions [13]. As an adrenergic antagonist, Carvedilol is known by its antioxidant and anti-inflammatory properties [14]. Increasing evidence has shown that Carvedilol is safely applied in the treatment of congestive heart failure, hypertension and coronary disease [15]. However, there is no report on the chondroprotective role of carvedilol before. Hence, this study addressed for the first time the

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