Protective effects of polydatin on lipopolysaccharide-induced acute lung injury through TLR4-MyD88-NF-κB pathway

doi:10.1016/j.intimp.2015.10.027

International Immunopharmacology

Volume 29, Issue 2, December 2015, Pages 370-376

https://doi.org/10.1016/j.intimp.2015.10.027 Get rights and content

Highlights

•
We first argued that polydatin could be a potential useful therapeutic agent for LPS-induced acute lung injury in mice.
•
We first explained that polydatin ameliorated LPS-induced acute lung injury through TLR4-MyD88-NF-κB pathway.
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Polydatin exhibited protective effects on LPS-induced ALI and BEAS-2B cells via TLR4-MyD88-NF-κB pathway.

Abstract

The purpose of this study was to investigate the protective effect of PD against lipopolysaccharide (LPS)-induced acute lung injury (ALI) and explore its potential mechanism. In vivo, PD and dexamethasone were intraperitoneally administered 1 h before LPS stimulation. Then, mice were sacrificed at 6 h post-LPS stimulation. Neutrophil number, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in bronchoalveolar lavage fluid (BALF) were determined, as well as lung wet to dry ratio (W/D) and polymorphonuclear (MPO) activity. The protein expressions of Toll like receptor 4 (TLR4), myeloid differentiating factor 88 (MyD88), IL-1R-associated kinases 1 (IRAK1), IRAK4, inhibitor of nuclear factor kappa-B kinase (IKK)α, p-IKKα, IKKβ, p-IKKβ, inhibitor of NF-κB (IκBα), p-IκBα and NF-κB in lung tissues were assessed. Besides, we detected the IL-6, IL-1β, IL-8, TNF-α levels and TLR4, MyD88, NF-κB protein expressions in LPS-induced BEAS-2B cells. Consequently, PD significantly inhibited the levels of W/D, MPO, neutrophils number, TNF-α, IL-6, IL-1β and reversed TLR4-MyD88-NF-κB signaling pathway in lung tissues. In vitro assays, PD effectively negatively mediated the inflammatory cytokines and ameliorated the high expressions of TLR4, MyD88, NF-κB caused by LPS simulation in Human bronchial epithelial BEAS-2B cells. This study indicated that PD played a protective role in LPS-induced ALI and BEAS-2B cells. The results supported further study of PD as potential candidate for acute lung injury.

Introduction

Acute lung injury is characteristic by hypoxemia, alveolar-capillary barrier damage, cells recruitment and a cascade of inflammatory process. Literatures have revealed that inflammatory cytokines such as IL-6, IL-1β and TNF-α are highly associated with ALI [1]. Despite the high morbidity and mortality, there has been few effective drug to treat ALI [2]. Recently, large quantities of investigations have proved that Toll like receptors (TLRs) play an essential role in innate immune system which is responsible for the inflammatory process in ALI. TLR family is composed of 10 members (TLR1-TLR10) [3], among which TLR4 can be engaged by LPS which is recognized as the component of outer membrane of Gram negative bacteria. TLR4 initiates a cascade of response including neutrophils infiltration and the accumulation of cytokines [4]. Classically, intratracheal administration of LPS is widely used as an inducer of lung injury in scientific studies.

It is well established that inflammatory process evoked by LPS is mediated by the transmembrane receptor toll-like receptor 4 (TLR4) [5]. After the recognition of LPS by TLR4, a serious of cascades including myeloid differentiating factor 88 (MyD88) are initiated following the activation of IRAKs. Subsequently, the recruitment of IRAK4 and IRAK1 to the MyD88-signaling complex leads to the activation of IKK-α and IKK-β, followed by the phosphorylated degradation of IκB-α and NF-κB initiation [6]. NF-κB, a regulator of inflammatory disorder, is required for the transcription of sufficient cytokines including IL-6, IL-1β, TNF-α [7]. In particular, the activation of NF-κB is attributed to TLR4 and is pivotal to the pathogenesis in various pulmonary diseases, such as ALI [8].

Traditional Chinese medicine has been used for centuries around the world and is still acknowledged as a major source of medicine [9], [10]. Resveratrol (3, 4′, 5-trihydroxy-trans-stilbene) is a non-flavonoid polyphenol and exerts anti-inflammatory and cardioprotective effects on human trials [11], [12]. Evidence emerged indicating that Resveratrol inhibited monocrotaline-induced pulmonary hypertension in rats [13]. Polydatin (PD), a crystal compound extracted from the root of Polygonum cuspidatum, is a glucoside of resveratrol in which the glucoside group bonded in position C-3 substitutes a hydroxyl group. The substitution contributes to the conformational changes of the molecule, which makes PD a potent molecule endowed with greater bioavailability respect to resveratrol [14]. PD is traditionally used for the treatment of chronic bronchitis, hepatitis, and shock [15]. Indraccolo et al. proved that PD showed anti-inflammatory properties during the inflammatory process of chronic pelvic pain in patients [16]. Whereas the pharmacological effects of PD on LPS-induced ALI in mice have not been elucidated. The purpose of this study was to assure the therapeutic efficacy of PD on ALI in mice with LPS challenge.

Section snippets

Reagents

Polydatin was provided by National Institutes for Food and Drug Control (purity more than 99.5%, Beijing, China) (Fig. 1) and dissolved in DMSO at a concentration of 200 mM. The final concentration of DMSO was less than 0.1% [v/v] in all experiments. Dexamethasone (DEX) was supplied by the National Institutes for Food and Drug Control (Beijing, China). LPS (E. coli 055:B5)) was purchased from Beyotime Institute of Biotechnology (Nanjing, China). Mouse IL-6, IL-1β, TNF-α enzyme-linked

Effects of PD on lung W/D ratio in LPS-induced ALI mice

The lung W/D (wet to dry) ratio was determined to reflect the pulmonary edema. As revealed in Fig. 1A, administration of LPS increased the level of lung W/D ratio. Notably, pretreatment with PD (80 mg/kg) significantly declined the magnitude of pulmonary edema, which was slightly less potent than those of pretreatment with dexamethasone. The experimental data suggested that PD could affect the water content of lung tissues in mice (Fig. 2A).

Effects of PD on MPO activity of the lung tissues in LPS-induced ALI mice

As an important measurement of neutrophils

Discussion

Polydatin (PD), isolated from P. cuspidatum, possesses multiple pharmacological actions. It has been reported to exhibit therapeutic effects in LPS-stimulated ALI in rats [17] and exert anti-inflammatory effects via NF-κB in RAW 264.7 cells [18]. However, few research had focused on the pharmacological effects of PD on ALI in LPS-induced mice and the explanation of its potential mechanism as yet.

LPS, the important constituent of the Gram-negative bacteria, is a major component of pathogens for

Conflict of interest

The authors have no conflict of interest.

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¹: These authors contributed equally and should be considered co-first-authors.

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Protective effects of polydatin on lipopolysaccharide-induced acute lung injury through TLR4-MyD88-NF-κB pathway

Highlights

Abstract

Introduction

Section snippets

Reagents

Effects of PD on lung W/D ratio in LPS-induced ALI mice

Effects of PD on MPO activity of the lung tissues in LPS-induced ALI mice

Discussion

Conflict of interest

J. Ethnopharmacol.

Int. Immunopharmacol.

Chem. Biol. Interact.

Int. Immunopharmacol.

Int. Immunopharmacol.

Pharmacol. Res.

Am. J. Cardiol.

Eur. J. Obstet. Gynecol. Reprod. Biol.

Int. Immunopharmacol.

Toxicol. Appl. Pharmacol.

Int. Immunopharmacol.

Int. Immunopharmacol.

Immunobiology

Behav. Brain Res.

Int. Immunopharmacol.

Eur. J. Pharmacol.

Hypoxia preconditioning increases survival and decreases expression of Toll-like receptor 4 in pulmonary artery endothelial cells exposed to lipopolysaccharide

Pulm Circ

Anti-inflammatory effects of apigenin in lipopolysaccharide-induced inflammatory in acute lung injury by suppressing COX-2 and NF-kB pathway

Inflammation

Esculin inhibits the inflammation of LPS-induced acute lung injury in mice via regulation of TLR/NF-kappaB pathways

Inflammation