特洛细胞在动脉粥样硬化小鼠颈动脉血管损伤修复中的变化

Changes of telocytes in the repair of carotid artery vascular injury in atherosclerotic mice

目的观察特洛细胞在动脉粥样硬化小鼠颈动脉血管损伤修复中的形态及数量变化。方法选取Apo E基因敲除小鼠30只作为观察组,给予高脂饮食喂养10周建立动脉粥样硬化模型;选取正常SPF级小鼠30只作为对照组,正常饮食喂养10周。两组均进行颈动脉锐性损伤后缝合,分别于术前及术后8 h、48 h、21 d麻醉后分离颈动脉标本。以CD34、CD117、血小板衍生生长因子受体(PDGFR)作为免疫标记,采用双标记免疫荧光染色法观察特洛细胞形态,并对10μm 2血管图像中的CD34阳性染色(CD34+)特洛细胞进行计数。结果观察组和对照组术前绿色荧光的CD34+和PDGFR+特洛细胞均集中位于血管壁的外膜层中。术后8 h,观察组CD34+特洛细胞聚集在血管壁外膜损伤处,细胞形态改变不明显。术后48 h,观察组CD117+特洛细胞数量明显减少,细胞体扁平,由圆形或椭圆形变为细长的梭形,端足细长如丝;对照组CD34+和CD117+特洛细胞形态较术后8 h无明显差别,端足形态无明显改变。术后21 d,观察组CD117+特洛细胞细胞恢复至术前和术后8 h时的形态,但数量较术后8 h明显减少;对照组CD34+、CD117+和PDGFR+特洛细胞形态较术前无明显差别,数量无明显改变。随着时间的推移,两组术后特洛细胞数量均呈现先升高后降低趋势。两组术前特洛细胞数量比较无明显差异(P>0.05)。观察组术后8 h特洛细胞数量高于术前,术后48 h和术后21 d特洛细胞数量均低于术前(P均<0.05);对照组术后8、48 h特洛细胞数量均高于术前(P均<0.05),术后21 d与术前比较无明显差异(P>0.05)。观察组术后8 h、48 h、21 d特洛细胞数量均低于对照组(P均<0.05)。结论动脉粥样硬化并不会影响小鼠颈动脉血管中的特洛细胞数量,但动脉粥样硬化小鼠在颈动脉血管损伤修复过程中存在特洛细胞数量降低及形态异常。

Objective To observe the morphological and quantitative changes of telocytes in carotid artery injury and repair in atherosclerotic mice.Methods Thirty Apo E knockout mice were selected as the observation group and fed a high-fat diet for 10 weeks to establish the atherosclerosis models,and meanwhile,30 SPF mice were selected as the control group and fed a normal diet for 10 weeks.The mice in both groups were sutured after acute carotid artery injury,and the carotid artery specimens were isolated before and after 8,48 h and 21 d,respectively.CD34,CD117,and platelet-derived growth factor receptor(PDGFR)were used as immunolabeling,and the morphology of telocytes was observed by double-labeled immunofluorescence staining,and CD34-positive staining(CD34+)telocytes were counted in 10μm 2 area of vascular images.Results CD34+and PDGFR+telocytes in both the observation and control groups were concentrated in the outer layer of the vessel wall.At 8 h after surgery,the CD34+and PDGFR+telocytes were concentrated in the outer membrane of the vessel wall,and the morphological changes of telocytes were not obvious.At 48 h,the number of CD117+telocytes decreased significantly,and the cell bodies were flattened,changing from round or oval to elongated shuttle-shaped,with elongated telopodes like filaments.There was no significant difference in the morphology of CD34+,CD117+and PDGFR+telopodes in the control group compared with the preoperative period and no significant change in the number of telopodes.There was no significant difference in the morphology of CD34+,CD117+and PDGFR+trophoblast cells in the control group as compared with those before operation.There was no significant difference in the number of telocytes between the two groups before surgery(P>0.05).In the control group,the number of telocytes at 8 and 48 h after surgery was higher than that before surgery(P>0.05),and the number of telocytes at 21 d after surgery was not significantly different from that before surgery(P>0.05).The number of telocytes was lower in the observation group than in the control group at 8,48 h and 21 d after surgery(all P<0.05).Conclusion Atherosclerosis does not affect the number of telocytes in the carotid arteries of mice,but there is a decrease in the number of telocytes and morphological abnormalities in the process of carotid artery injury and repair in atherosclerotic mice.