中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2015年
11期
770-773
,共4页
吴凡%张瑞丽%张津萍%王千秋
吳凡%張瑞麗%張津萍%王韆鞦
오범%장서려%장진평%왕천추
苍白密螺旋体%内皮细胞%微血管%人脑微血管内皮细胞
蒼白密螺鏇體%內皮細胞%微血管%人腦微血管內皮細胞
창백밀라선체%내피세포%미혈관%인뇌미혈관내피세포
Treponema pallidum%Endothelial cells%Microvessels%Human brain microvascular endothelial cells
目的 观察梅毒螺旋体对人脑微血管内皮细胞(HBMEC)的粘附.方法 将HBMEC接种于24孔板玻片中,加入1.6×107条/ml的梅毒螺旋体悬液混合培养,分别于培养0.5、2、4h,运用扫描电镜检查梅毒螺旋体与HBMEC的粘附方式;加不同密度(4×106条/ml、8×106条/ml、1.6×107/条ml)梅毒螺旋体悬液混合培养,于不同的时间点(2、4、6、16h)运用暗视野显微镜检查计数单个HBMEC上粘附的梅毒螺旋体数量.采用重复测量资料的方差分析对实验数据进行分析.结果 扫描电镜结果显示,梅毒螺旋体与HBMEC粘附时表现为集中粘附于HBMEC膜表面的某一区域,且在粘附的部位两者发生部分融合.加入不同浓度的梅毒螺旋体悬液与HBMEC混合培养后,不同时间点细胞上粘附的梅毒螺旋体数目差异有统计学意义(F=387.72,P<0.001),单个细胞上的梅毒螺旋体数量随着混合培养时间的延长而逐渐增多,6h时达到高峰,然后呈下降趋势,在16h时为最低.在各观察时间点,不同密度组细胞上粘附的梅毒螺旋体数目差异也有统计学意义(F=593.23,P<0.001),时间与密度存在交互作用(F=98.74,P< 0.001).结论 梅毒螺旋体可以粘附于体外培养的HBMEC,部分梅毒螺旋体可能通过末端与HBMEC膜溶解粘附于细胞表面.
目的 觀察梅毒螺鏇體對人腦微血管內皮細胞(HBMEC)的粘附.方法 將HBMEC接種于24孔闆玻片中,加入1.6×107條/ml的梅毒螺鏇體懸液混閤培養,分彆于培養0.5、2、4h,運用掃描電鏡檢查梅毒螺鏇體與HBMEC的粘附方式;加不同密度(4×106條/ml、8×106條/ml、1.6×107/條ml)梅毒螺鏇體懸液混閤培養,于不同的時間點(2、4、6、16h)運用暗視野顯微鏡檢查計數單箇HBMEC上粘附的梅毒螺鏇體數量.採用重複測量資料的方差分析對實驗數據進行分析.結果 掃描電鏡結果顯示,梅毒螺鏇體與HBMEC粘附時錶現為集中粘附于HBMEC膜錶麵的某一區域,且在粘附的部位兩者髮生部分融閤.加入不同濃度的梅毒螺鏇體懸液與HBMEC混閤培養後,不同時間點細胞上粘附的梅毒螺鏇體數目差異有統計學意義(F=387.72,P<0.001),單箇細胞上的梅毒螺鏇體數量隨著混閤培養時間的延長而逐漸增多,6h時達到高峰,然後呈下降趨勢,在16h時為最低.在各觀察時間點,不同密度組細胞上粘附的梅毒螺鏇體數目差異也有統計學意義(F=593.23,P<0.001),時間與密度存在交互作用(F=98.74,P< 0.001).結論 梅毒螺鏇體可以粘附于體外培養的HBMEC,部分梅毒螺鏇體可能通過末耑與HBMEC膜溶解粘附于細胞錶麵.
목적 관찰매독라선체대인뇌미혈관내피세포(HBMEC)적점부.방법 장HBMEC접충우24공판파편중,가입1.6×107조/ml적매독라선체현액혼합배양,분별우배양0.5、2、4h,운용소묘전경검사매독라선체여HBMEC적점부방식;가불동밀도(4×106조/ml、8×106조/ml、1.6×107/조ml)매독라선체현액혼합배양,우불동적시간점(2、4、6、16h)운용암시야현미경검사계수단개HBMEC상점부적매독라선체수량.채용중복측량자료적방차분석대실험수거진행분석.결과 소묘전경결과현시,매독라선체여HBMEC점부시표현위집중점부우HBMEC막표면적모일구역,차재점부적부위량자발생부분융합.가입불동농도적매독라선체현액여HBMEC혼합배양후,불동시간점세포상점부적매독라선체수목차이유통계학의의(F=387.72,P<0.001),단개세포상적매독라선체수량수착혼합배양시간적연장이축점증다,6h시체도고봉,연후정하강추세,재16h시위최저.재각관찰시간점,불동밀도조세포상점부적매독라선체수목차이야유통계학의의(F=593.23,P<0.001),시간여밀도존재교호작용(F=98.74,P< 0.001).결론 매독라선체가이점부우체외배양적HBMEC,부분매독라선체가능통과말단여HBMEC막용해점부우세포표면.
Objective To observe the attachment of Treponema pallidum to human brain microvascular endothelial cells (HBMECs) in vitro.Methods Some primary cultured HBMECs were inoculated into in 24-well plates to be cocultured with the suspension of T.pallidum at a concentration of 1.6 × 107 treponemes/ml.After 0.5,2 and 4 hours of co-culture,scanning electron microscopy was conducted to observe the attachment of T.pallidum to HBMECs.Some HBMECs were cocultured with the presence of T.pallidum suspensions at different concentrations (4 × 106,8 × 106,1.6 × 107 treponemes/ml) for 2,4,6 and 16 hours,then,dark-field microscopy was performed to count the number of treponemes that attached to single HBMECs.Statistical analysis was carried out by using repeated-measures analysis of variance.Results As scanning electron microscopy showed,treponemes gathered at some regions on the surface of HBMECs when they attached to HBMECs.In addition,T.pallidum partly merged with the membrane of HBMECs at the site of attachment.After co-culture with T.pallidum suspensions,the number of treponemes that attached to single HBMECs was significantly different among different time points (F =387.72,P < 0.001) and among different concentrations of T.pallidum suspensions (F =593.23,P < 0.001),with an interaction effect between the concentration of T.pallidum suspensions and incubation period (F =98.74,P < 0.001).Concretely speaking,the number of treponemes that attached to single HBMECs increased over time until 6 hours after the start of coculture,then showed a decreasing trend,and reached the nadir value at 16 hours.Conclusion T.pallidum can adhere to cultured HBMECs in vitro,likely by the merger of its end with the membrane of HBMECs at some regions.