血管紧张素Ⅱ对人牙周膜成纤维细胞活化作用的研究
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摘要
目的
通过AngⅡ作用于人牙周膜成纤维细胞后牙周膜成纤维细胞的一系列活化作用,探讨AngⅡ对人牙周膜成纤维细胞生物学特性的影响,为临床应用提供理论依据。
方法
(1)通过组织块培养法对人牙周膜成纤维细胞进行体外培养、传代及鉴定。
(2)通过细胞计数及MTT法测定AngⅡ对牙周膜成纤维细胞增殖的影响。
(3)建立牙周创伤模型,测定AngⅡ对牙周膜成纤维细胞迁移的影响,并通过photoshop软件分析。
(4)通过细胞免疫荧光技术测定AngⅡ对细胞合成Ⅰ型胶原的影响,并通过IPP图像分析软件进行半定量分析。
结果
(1)与对照组相比,AngⅡ能够明显促进牙周膜成纤维细胞的增殖(P＜0.05),该作用能被AT1受体拮抗剂Losartan阻断。
(2)与对照组相比,AngⅡ能够明显促进牙周膜成纤维细胞的迁移(P＜0.05)。
(3)AngⅡ能够明显促进牙周膜成纤维细胞合成Ⅰ型胶原(P＜0.05)。
结论
(1)牙周膜成纤维细胞是AngⅡ作用的靶细胞之一。
(2)AngⅡ能够诱导牙周膜成纤维细胞的增殖,迁移及Ⅰ型胶原的合成。AT1受体拮抗剂能够阻断该作用。
Objective
The identification and the development of potential growth factors for the cells of the periodontal ligament is an important and novel stratagem for the regeneration of tissue destroyed by periodontitis and periapical inflammation. The problem is that growth factors are difficult to synthesize, very immunogenic and expensive. The purpose of our study was to determine whether angiotensin II, a naturally occurring octo-peptide with growth factor like properties in cardiac muscle, could also stimulate cultured human periodontal ligament cells.
Methods
(1) Periodontal ligament cells were cultured and identified in vitro by explant method.
(2)Cell counting and MTT assay were used to study the effects of angiotensin II on the proliferation and activity of cultured cells.
(3)A wounding periodontal ligament cell model in vitro was established. The distance of cells migration was observed and measured by cultured with angiotensin II and losartan and analysed by photoshop software.
(4)Collagen I synthesis of periodontal ligament cells was determined by immunofluorescence technique. The semi-quantitative analysis was made to collagen by image pattern analysisor.
Results
(1)Angiotensin II can promote the proliferation and activation of periodontal ligament cells through AT1 receptor. The effect can be blocked by losartan.
(2)The cells cultured with angiotensin II migrated faster than those in common cultured or cultured with both losartan and angiotensin II.
(3) Angiotensin II can promote the collagen I synthesis. Losartan can block this effect.
Conclusions
(1) Periodontal ligament cell is the target cell of angiotensin II.
(2)Angiotensin II can promote the proliferation, migration and collagen I synthesis of periodontal ligament cells. The effects can be blocked by losartan.
通过AngⅡ作用于人牙周膜成纤维细胞后牙周膜成纤维细胞的一系列活化作用,探讨AngⅡ对人牙周膜成纤维细胞生物学特性的影响,为临床应用提供理论依据。
方法
(1)通过组织块培养法对人牙周膜成纤维细胞进行体外培养、传代及鉴定。
(2)通过细胞计数及MTT法测定AngⅡ对牙周膜成纤维细胞增殖的影响。
(3)建立牙周创伤模型,测定AngⅡ对牙周膜成纤维细胞迁移的影响,并通过photoshop软件分析。
(4)通过细胞免疫荧光技术测定AngⅡ对细胞合成Ⅰ型胶原的影响,并通过IPP图像分析软件进行半定量分析。
结果
(1)与对照组相比,AngⅡ能够明显促进牙周膜成纤维细胞的增殖(P＜0.05),该作用能被AT1受体拮抗剂Losartan阻断。
(2)与对照组相比,AngⅡ能够明显促进牙周膜成纤维细胞的迁移(P＜0.05)。
(3)AngⅡ能够明显促进牙周膜成纤维细胞合成Ⅰ型胶原(P＜0.05)。
结论
(1)牙周膜成纤维细胞是AngⅡ作用的靶细胞之一。
(2)AngⅡ能够诱导牙周膜成纤维细胞的增殖,迁移及Ⅰ型胶原的合成。AT1受体拮抗剂能够阻断该作用。
Objective
The identification and the development of potential growth factors for the cells of the periodontal ligament is an important and novel stratagem for the regeneration of tissue destroyed by periodontitis and periapical inflammation. The problem is that growth factors are difficult to synthesize, very immunogenic and expensive. The purpose of our study was to determine whether angiotensin II, a naturally occurring octo-peptide with growth factor like properties in cardiac muscle, could also stimulate cultured human periodontal ligament cells.
Methods
(1) Periodontal ligament cells were cultured and identified in vitro by explant method.
(2)Cell counting and MTT assay were used to study the effects of angiotensin II on the proliferation and activity of cultured cells.
(3)A wounding periodontal ligament cell model in vitro was established. The distance of cells migration was observed and measured by cultured with angiotensin II and losartan and analysed by photoshop software.
(4)Collagen I synthesis of periodontal ligament cells was determined by immunofluorescence technique. The semi-quantitative analysis was made to collagen by image pattern analysisor.
Results
(1)Angiotensin II can promote the proliferation and activation of periodontal ligament cells through AT1 receptor. The effect can be blocked by losartan.
(2)The cells cultured with angiotensin II migrated faster than those in common cultured or cultured with both losartan and angiotensin II.
(3) Angiotensin II can promote the collagen I synthesis. Losartan can block this effect.
Conclusions
(1) Periodontal ligament cell is the target cell of angiotensin II.
(2)Angiotensin II can promote the proliferation, migration and collagen I synthesis of periodontal ligament cells. The effects can be blocked by losartan.
引文
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[4]张艺军,杨希山,吴平生等.血管紧张素Ⅱ和洛沙坦对肝星状细胞合成胶原的影响.中国病理生理杂志,2004,20(10):1842-1845.
[5]龙翔,熊盛道,熊维宁等.氯沙坦对肺成纤维细胞转分化和胶原合成的抑制作用.中华结核和呼吸杂志,2007,30(4):273-277.
[6]陈艳清,李世荣,曹川等.血管紧张素Ⅱ对增生性瘢痕成纤维细胞生长增殖的影响.中国美容医学,2007,16(3):298-301.
[7]Antoniu SA.Targeting the angiotensin pathway in idiopathic pulmonary fibrosis.Expert Opin Ther Targets,2008,12(12):1587-1590.
[8]Lundergan WP,Ferry D,Kobayashi H,et al.Angiotensin-Ⅱ increases cytoplasmic calcium,cell number and total DNA for human periodontal ligamental cells in vitro.J Periodont Res,1999,34:223-228.
[9]司徒镇强,吴军正.细胞培养.北京:世界出版社,2007.69-70.
[10]王健平,隋晓梅.组织块法和酶消化法培养人牙周膜细胞.黑龙江医药科学,2002,25(6):4-6.
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