title: [Hydroxyfasudi attenuates lipopolysaccharide induced endothelial dysfunction via suppressing the expression of Rho-associated coiled-coil protein kinase 1, connexin 43 and caveolin 1].
abstract: Objective:To investigate the effect and mechanism of hydroxyfasudi (HF), a specific Rho kinase inhibitor, on lipopolysaccharide(LPS)induced endothelial dysfunction.Methods:A total of 24 male Sprague Dawley rats were randomly divided into control group(n=6), HF group(n=6), LPS group(n=6) and LPS + HF group(n=6) with random number table method. There was no special treatment in control group. HF (30 mg/kg) was injected intraperitoneally in HF group. LPS (1 mg/kg) were injected intravenously in LPS group. In LPS+ HF group, HF (30 mg/kg) was injected intraperitoneally, followed by intravenous LPS injection (1 mg/kg) 30 minutes later. All rats were sacrificed after 8 hours, and aortic tissue was extracted. RT-PCR was performed to detect mRNA levels of Rho-associated coiled-coil protein kinase (ROCK)1, connexin (Cx)43 and caveolin (Cav)1. The protein levers of ROCK1, Cx43 and Cav-1 were assessed by Western blot and immunohistochemical staining respectively.Results:(1) RT-PCR experiments showed that mRNA levels of ROCK1(2.67±0.03 vs. 1.00±0.04), Cx43(1.73±0.03 vs. 1.00±0.08), and Cav1(1.85±0.04 vs. 1.0±0.03) in LPS group were significantly higher than in control group(allP<0.05). mRNA levels of ROCK1(0.38±0.02), Cx43(0.58±0.02), and Cav1(0.27±0.01) in LPS + HF group were significantly lower than in LPS group(allP<0.05). (2)Western blot analysis showed that protein levels of ROCK1(3.46±0.82 vs. 2.19±0.56), Cx43(0.33±0.09 vs.0.11±0.06), and Cav1(3.45±0.74 vs. 2.25±0.91) in LPS group were significantly higher than in control group(allP<0.05). Protein levels of ROCK1(1.09±0.52), Cx43(0.01±0.06), and Cav1(2.06±0.40) in LPS + HF group were significantly lower than in LPS group(allP<0.05). (3) Immunohistochemical staining showed that protein levels of ROCK1(84.1±0.953.7±2.9), Cx43(99.1±2.1 vs. 46.2±0.8), and Cav1(167.0±6.4 vs. 84.9±1.0) in LPS group were significantly higher than in control group(allP<0.05). Protein levels of ROCK1(30.4±0.6), Cx43(21.4±1.3), and Cav1(55.8±2.8) in LPS + HF group were significantly lower than in LPS group(allP<0.05).Conclusion:HF attenuates LPS induced endothelial dysfunction probably via suppressing the expression of ROCK1, Cx43 and Cav1.
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Guangxi Medical University, Nanning 530021, China.
authors.forename: P F
authors.initials: PF
authors.lastname: Zheng
authors.forename: X H
authors.initials: XH
authors.lastname: Wu
authors.forename: W
authors.initials: W
authors.lastname: Huang
authors.forename: G T
authors.initials: GT
authors.lastname: Ma
authors.forename: Q
authors.initials: Q
authors.lastname: Bi
mesh_terms: D000818:Animals; D051242:Caveolin 1; D018031:Connexin 43; D004727:Endothelium; D008070:Lipopolysaccharides; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D054460:rho-Associated Kinases
chemical_list: D051242:Caveolin 1; D018031:Connexin 43; C580592:Gja1 protein, rat; D008070:Lipopolysaccharides; C578418:ROCK1 protein, rat; D054460:rho-Associated Kinases
keywords: Connexins; Endothelium, vascular; Hydroxyfasudi