Objective:

2

To examine the anti-inflammatory effects and potential mechanisms of polypeptide from

Moschus

(PPM) in lipopolysaccharide (LPS)-induced THP-1 macrophages and BALB/c mice.

Methods:

2

The polypeptide was extracted from

Moschus

and analyzed by high-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Subsequently

LPS was used to induce inflammation in THP-1 macrophages and BALB/c mice. In LPS-treated or untreated THP-1 macrophages

cell viability was observed by cell counting kit 8 and lactate dehydrogenase release assays; the proinflammatory cytokines and reactive oxygen species (ROS) were measured by enzyme-linked immunosorbent assay and flow cytometry

respectively; and protein and mRNA levels were measured by Western blot and real-time quantitative polymerase chain reaction (qRT-PCR)

respectively. In LPS-induced BALB/c mice

the proinflammatory cytokines were measured

and lung histology and cytokines were observed by hematoxylin and eosin (HE) and immunohistochemical (IHC) staining

respectively.

Results:

2

The SDS-PAGE results suggested that the molecular weight of purified PPM was in the range of 10–26 kD.

In vitro

PPM reduced the production of interleukin 1β (IL-1β)

IL-18

tumor necrosis factor α (TNF-α)

IL-6 and ROS in LPS-induced THP-1 macrophages (

P

<

0.01). Western blot analysis demonstrated that PPM inhibited LPS-induced nuclear factor κB (NF-κB) pathway and thioredoxin interacting protein (TXNIP)/nucleotide-binding oligomerization domain

leucine-rich repeat and pyrin domain containing 3 (NLRP3) inflammasome pathway by reducing protein expression of phospho-NF-κB p65

phospho- inhibitors of NF-κB (IκBs) kinase α/β (IKKα/β)

TXNIP

NLRP3

apoptosis-associated speck- like protein containing a caspase recruitment domain (ASC)

and pro-caspase-1 (

P

<

0.05 or

P

<

0.01). In addition

qRT-PCR revealed the inhibitory effects of PPM on the mRNA levels of TXNIP

NLRP3

ASC

and caspase-1 (

P

<

0.05 or

P

<

0.01). Furthermore

in LPS-induced BALB/c mice

PPM reduced TNF-α and IL-6 levels in serum (

P

<

0.05 or

P

<

0.01)

decreased IL-1β and IL-18 levels in the lungs (

P

<

0.01) and alleviated pathological injury to the lungs.

Conclusion:

2

PPM could attenuate LPS-induced inflammation by inhibiting the NF-κB-ROS/NLRP3 pathway

and may be a novel potential candidate drug for treating inflammation and inflammation-related diseases.