title: Gingipains from Porphyromonas gingivalis promote the transformation and proliferation of vascular smooth muscle cell phenotypes.
abstract: The aim of the present study was to ascertain the effect of Porphyromonas gingivalis cysteine protease gingipain on the proliferation of rat aortic smooth muscle cells (RASMCs). Gingipains were isolated and purified from the supernatant of P. gingivalis W83, which was cultured under standard anaerobic conditions; primary RASMCs were also cultured. RASMCs were exposed to 200, 100, 50, 25, 12, 6, 3, 1, and 0 μg/mL activated gingipains and the proliferation was evaluated using a cell counting kit-8 (CCK-8) assay after 48 h. α-Smooth muscle actin (α-SMA) and osteopontin (OPN) expression were measured by immunohistochemical staining. In addition, RASMCs were stimulated with 5, 10, 20, and 40 μM KYT-1 (arg-gingipain inhibitor) and KYT-36 (lys-gingipain inhibitor) in combination with the gingipain extracts. Different concentrations of gingipains significantly promoted the proliferation of RASMCs, except those treated with 1 μg/mL, compared to the untreated controls. The proliferation was sustained at a concentration above 12 μg/mL. Immunohistochemical staining showed OPN expression after gingipain stimulation. The proliferative effects of gingipains on RASMCs were blocked after treatment with 10 μM KYT-1 or 10 μM KYT-36 (P < 0.0001); however, the difference between KYT-1 and KYT-36 groups was not statistically significant. These results demonstrated that gingipains can promote phenotypic transformation and proliferation of RASMCs and their effects were blocked by KYT-1 and KYT-36, which help us to ascertain whether Rgp or Kgp contributes to the development of atherosclerosis.
authors.affiliation: Department of Periodontology, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, China.
authors.forename: Chong
authors.initials: C
authors.lastname: Cao
authors.affiliation: Department of Prosthodontics, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, China.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Ji
authors.affiliation: Department of Pharmacology, The Basic Medical Sciences College of Xinjiang Medical University Urumqi 830054, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Luo
authors.affiliation: Department of Periodontology, The First Affiliated Hospital of Xinjiang Medical UniversityUrumqi 830054, China; Medical College of Hangzhou Normal UniversityHangzhou 311121, China; Department of Stomatology, The Affiliated Hospital of Hangzhou Normal UniversityHangzhou 310015, China.
authors.forename: Liangjun
authors.initials: L
authors.lastname: Zhong
keywords: Gingipains; phenotype transformation; proliferation; smooth muscle cell