title: Label-free LC-MS/MS shotgun proteomics to investigate the anti-inflammatory effect of rCC16.
abstract: Clara cell protein (CC16) is an anti-inflammatory protein, which is expressed in the airway epithelium. It is involved in the development of airway inflammatory diseases, including chronic obstructive pulmonary disease and asthma. However, the exact molecular mechanism underlying its anti‑inflammatory action remains to be fully elucidated. The aim of the present study was to define the protein profiles of the anti‑inflammatory effect of CC16 in lipopolysaccharide (LPS)‑treated rat tracheal epithelial (RTE) cells using shotgun proteomics. Protein extracts were obtained from control RTE cells, RTE cells treated with LPS and RTE cells treated with LPS and recombinant CC16 (rCC16). Subsequent label‑free quantification and bioinformatics analyses identified 12 proteins that were differentially expressed in the three treatment groups as a cluster of five distinct groups according to their molecular functions. Five of the twelve proteins were revealed to be associated with the cytoskeleton: Matrix metalloproteinase‑9, myosin heavy chain 10, actin‑related protein‑3 homolog, elongation factor 1‑α‑1 (EF‑1‑α‑1), and acidic ribosomal phosphoprotein P0. Five of the twelve proteins were associated with cellular proliferation: DNA‑dependent protein kinase catalytic subunit, EF‑1‑α‑1, tyrosine 3‑monooxygenase, caspase recruitment domain (CARD) protein 12 and adenosylhomocysteinase (SAHH) 3. Three proteins were associated with gene regulation: EF‑1‑α‑1, SAHH 3 and acidic ribosomal phosphoprotein P0. Three proteins were associated with inflammation: Tyrosine 3‑monooxygenase, CARD protein 12 and statin‑related protein. ATPase (H+‑transporting, V1 subunit A, isoform 1) was revealed to be associated with energy metabolism, and uridine diphosphate glycosyltransferase 1 family polypeptide A8 with drug metabolism and detoxification. The identified proteins were further validated using reverse transcription‑quantitative polymerase chain reaction. These protein profiles, and their interacting protein network, may facilitate the elucidation of the molecular mechanisms underlying the anti‑inflammatory effects of CC16.
authors.affiliation: Respiratory Department, The First Affiliated Hospital, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China.
authors.forename: Min
authors.initials: M
authors.lastname: Pang
authors.affiliation: Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China.
authors.forename: Xin-Yan
authors.initials: XY
authors.lastname: Bai
authors.affiliation: Fan‑Xing Biological Technology Co., Ltd., Beijing 010000, P.R. China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Physiology, Faculty of Medical and Health Sciences, University of Auckland, Auckland 1142, New Zealand.
authors.forename: Ji-Zhong
authors.initials: JZ
authors.lastname: Bai
authors.affiliation: Respiratory Department, The First Affiliated Hospital, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China.
authors.forename: Li-Rong
authors.initials: LR
authors.lastname: Yuan
authors.affiliation: Respiratory Department, The First Affiliated Hospital, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China.
authors.forename: Shou-An
authors.initials: SA
authors.lastname: Ren
authors.affiliation: Respiratory Department, The First Affiliated Hospital, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China.
authors.forename: Xiao-Yun
authors.initials: XY
authors.lastname: Hu
authors.affiliation: Respiratory Department, The First Affiliated Hospital, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China.
authors.forename: Xin-Ri
authors.initials: XR
authors.lastname: Zhang
authors.affiliation: Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China.
authors.forename: Bao-Feng
authors.initials: BF
authors.lastname: Yu
authors.affiliation: Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China.
authors.forename: Rui
authors.initials: R
authors.lastname: Guo
authors.affiliation: Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China.
authors.forename: Hai-Long
authors.initials: HL
authors.lastname: Wang
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007249:Inflammation / Q000139:chemically induced / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D008070:Lipopolysaccharides / Q000633:toxicity; D014176:Protein Biosynthesis / Q000235:genetics; D040901:Proteomics; D029424:Pulmonary Disease, Chronic Obstructive; D051381:Rats; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics* / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D014132:Trachea / Q000378:metabolism* / Q000473:pathology; D014598:Uteroglobin / Q000008:administration & dosage / Q000235:genetics* / Q000378:metabolism
chemical_list: D008070:Lipopolysaccharides; D011994:Recombinant Proteins; C556391:Scgb1a1 protein, rat; D014598:Uteroglobin