Du Yang1, Yuan Shuai1, Zhou Zhifei1, Wu Lizheng2, Wang Lulu1, Wu Xing'an3, Wang Xiaojing1. 1. State Key Laboratory of Military Stomatology, National Clinical Research Center for Oral Diseases, Shanxi Clinical Research Center for Oral Diseases, Dept. of Pediatric Dentistry, School of Stomatology, The Fourth Military Medical University, Xi'an 710032, China. 2. Dept. of Stomatology, Affiliated Hospital of Logistics University of People's Armed Police Force, Tianjin 300300, China. 3. Dept. of Microbiology, The Fourth Military Medical University, Xi'an 710032, China.
Abstract
OBJECTIVE: To explore the effect of nicotine on the autophagy level of human periodontal ligament cells (hPDLCs). METHODS: Periodontal tissues collected from premolars for orthodontic treatment reasons were used to culture hPDLCs. Western blot analysis was performed to test the most optimal time and concentration of nicotine on the autophagy level of the hPDLCs. Transmission electron microscope and immunofluorescence observation were carried out to detect the form of autophagosomes and expression of autophagy related protein LC3 in hPDLCs under this optimal condition. RESULTS: Protein expression of LC3Ⅱ was up regulated with the 12 h nicotine stimulating. Besides that, the up regulation of the protein expression of LC3Ⅱ was concentration dependent and nicotine with a concentration of 1×10⁻⁵ mol·L⁻¹ was the most optimal condition. Transmission electron microscope and immunofluorescence observations indicated that nicotine would activate the autophagy level of hPDLCs by increasing the number of autophagosomes and up regulating the expression of autophagy related protein LC3. CONCLUSIONS: Nicotine could increase autophagy level of hPDLCs, thus affecting the occurrence and development of smoking related periodontitis.
OBJECTIVE: To explore the effect of nicotine on the autophagy level of human periodontal ligament cells (hPDLCs). METHODS: Periodontal tissues collected from premolars for orthodontic treatment reasons were used to culture hPDLCs. Western blot analysis was performed to test the most optimal time and concentration of nicotine on the autophagy level of the hPDLCs. Transmission electron microscope and immunofluorescence observation were carried out to detect the form of autophagosomes and expression of autophagy related protein LC3 in hPDLCs under this optimal condition. RESULTS: Protein expression of LC3Ⅱ was up regulated with the 12 h nicotine stimulating. Besides that, the up regulation of the protein expression of LC3Ⅱ was concentration dependent and nicotine with a concentration of 1×10⁻⁵ mol·L⁻¹ was the most optimal condition. Transmission electron microscope and immunofluorescence observations indicated that nicotine would activate the autophagy level of hPDLCs by increasing the number of autophagosomes and up regulating the expression of autophagy related protein LC3. CONCLUSIONS:Nicotine could increase autophagy level of hPDLCs, thus affecting the occurrence and development of smoking related periodontitis.
Entities:
Keywords:
autophagy; human periodontal ligament cells; nicotine
Authors: Pedro Bullon; Mario David Cordero; José Luis Quiles; Maria Del Carmen Ramirez-Tortosa; Adrian Gonzalez-Alonso; Simona Alfonsi; Rocio García-Marín; Manuel de Miguel; Maurizio Battino Journal: BMC Med Date: 2012-10-17 Impact factor: 8.775