PMID- 36178087 OWN - NLM STAT- MEDLINE DCOM- 20221003 LR - 20230126 IS - 2001-1326 (Electronic) IS - 2001-1326 (Linking) VI - 12 IP - 10 DP - 2022 Oct TI - Inhibition of ROCK ameliorates pulmonary fibrosis by suppressing M2 macrophage polarisation through phosphorylation of STAT3. PG - e1036 LID - 10.1002/ctm2.1036 [doi] LID - e1036 AB - BACKGROUND: Emerging evidence provides mechanistic insights into the pathogenesis of pulmonary fibrosis (PF), and rare anti-PF therapeutic method has promising effect in its treatment. Rho-associated coiled-coil kinases (ROCK) inhibition significantly ameliorates bleomycin-induced PF and decreases macrophage infiltration, but the mechanism remains unclear. We established bleomycin and radiation-induced PF to identify the activity of WXWH0265, a newly designed unselective ROCK inhibitor in regulating macrophages. METHODS: Bleomycin-induced PF was induced by intratracheal instillation and radiation-induced PF was induced by bilateral thoracic irradiation. Histopathological techniques (haematoxylin and eosin, Masson's trichrome and immunohistochemistry) and hydroxyproline were used to evaluate PF severity. Western blot, quantitative real-time reverse transcription-polymerase chain reaction and flow cytometry were performed to explore the underlying mechanisms. Bone marrow-derived macrophages (BMDMs) were used to verify their therapeutic effect. Clodronate liposomes were applied to deplete macrophages and to identify the therapeutic effect of WXWH0265. RESULTS: Therapeutic administration of ROCK inhibitor ameliorates bleomycin-induced PF by inhibiting M2 macrophages polarisation. ROCK inhibitor showed no significant anti-fibrotic effect in macrophages-depleted mice. Treatment with WXWH0265 demonstrated superior protection effect in bleomycin-induced PF compared with positive drugs. In radiation-induced PF, ROCK inhibitor effectively ameliorated PF. Fibroblasts co-cultured with supernatant from various M2 macrophages phenotypes revealed that M2 macrophages stimulated by interleukin-4 promoted extracellular matrix production. Polarisation of M2 macrophages was inhibited by ROCK inhibitor treatment in vitro. The p-signal transducer and activator of transcription 3 (STAT3) in lung tissue and BMDMs was significantly decreased in PF in vivo and vitro after treated with ROCK inhibitors. CONCLUSION: Inhibiting ROCK could significantly attenuate bleomycin- and radiation-induced PF by regulating the macrophages polarisation via phosphorylation of STAT3. WXWH0265 is a kind of efficient unselective ROCK inhibitor in ameliorating PF. Furthermore, the results provide empirical evidence that ROCK inhibitor, WXWH0265 is a potential drug to prevent the development of PF. CI - (c) 2022 The Authors. Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics. FAU - Li, Qingfang AU - Li Q AUID- ORCID: 0000-0001-7671-8973 AD - Laboratory of Aging Research and Cancer Drug Target, State Key Laboratory of Biotherapy, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, Sichuan, PR China. FAU - Cheng, Yuan AU - Cheng Y AD - Laboratory of Aging Research and Cancer Drug Target, State Key Laboratory of Biotherapy, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, Sichuan, PR China. FAU - Zhang, Zhe AU - Zhang Z AD - Laboratory of Aging Research and Cancer Drug Target, State Key Laboratory of Biotherapy, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, Sichuan, PR China. FAU - Bi, Zhenfei AU - Bi Z AD - Laboratory of Aging Research and Cancer Drug Target, State Key Laboratory of Biotherapy, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, Sichuan, PR China. FAU - Ma, Xuelei AU - Ma X AD - Laboratory of Aging Research and Cancer Drug Target, State Key Laboratory of Biotherapy, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, Sichuan, PR China. FAU - Wei, Yuquan AU - Wei Y AD - Laboratory of Aging Research and Cancer Drug Target, State Key Laboratory of Biotherapy, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, Sichuan, PR China. FAU - Wei, Xiawei AU - Wei X AUID- ORCID: 0000-0002-6513-6422 AD - Laboratory of Aging Research and Cancer Drug Target, State Key Laboratory of Biotherapy, National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, Sichuan, PR China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - United States TA - Clin Transl Med JT - Clinical and translational medicine JID - 101597971 RN - 0 (Liposomes) RN - 0 (STAT3 Transcription Factor) RN - 0813BZ6866 (Clodronic Acid) RN - 11056-06-7 (Bleomycin) RN - 207137-56-2 (Interleukin-4) RN - EC 2.7.11.1 (rho-Associated Kinases) SB - IM MH - Animals MH - Bleomycin/adverse effects MH - Clodronic Acid/metabolism MH - Interleukin-4/metabolism MH - Liposomes/metabolism MH - Macrophages/metabolism MH - Mice MH - Phosphorylation MH - *Pulmonary Fibrosis/chemically induced/drug therapy/pathology MH - *STAT3 Transcription Factor/metabolism MH - *rho-Associated Kinases/antagonists & inhibitors PMC - PMC9523675 OTO - NOTNLM OT - idiopathic pulmonary fibrosis OT - macrophage OT - polarisation OT - radiation-induced pulmonary fibrosis COIS- The authors declare they have no conflicts of interest. EDAT- 2022/10/01 06:00 MHDA- 2022/10/04 06:00 PMCR- 2022/09/30 CRDT- 2022/09/30 06:44 PHST- 2022/08/09 00:00 [revised] PHST- 2022/01/23 00:00 [received] PHST- 2022/08/15 00:00 [accepted] PHST- 2022/09/30 06:44 [entrez] PHST- 2022/10/01 06:00 [pubmed] PHST- 2022/10/04 06:00 [medline] PHST- 2022/09/30 00:00 [pmc-release] AID - CTM21036 [pii] AID - 10.1002/ctm2.1036 [doi] PST - ppublish SO - Clin Transl Med. 2022 Oct;12(10):e1036. doi: 10.1002/ctm2.1036.