内毒素和内皮细胞的结合能力及对a-Catenin的影响

Combinative Ability of Endotoxin With Cultured Endothelial Cells and Effects of Endotoxin on Endothelial Intracellularα-Catenin

寻找内毒素和培养的内皮细胞直接结合的证据及对胞内连接蛋白α－Ｃａｔｅｎｉｎ的影响，观察内毒素对内皮细胞的损伤特性。方法：（１）荧光标记的内毒素（２μｍ／ｍｌ）和培养的内皮细胞在无血清下共同孵育后用荧光分光光度计进行检测；（２）内毒素（２μｇ／ｍｌ）刺激无血清培养的内皮细胞后用ＥＬＩＳＡ测定内皮细胞胞内连接蛋白α－Ｃａｔｅｎｉｎ表达的变化。结果：内皮细胞和内毒素孵育１０、２０、４０和６０分后荧光强度分别为对照组的１．８７、３．９７、３．４１（Ｐ＜０．０１）和２．０２（Ｐ＜０．０５）倍；胞内连接蛋白的表达各时相点和对照组比均明显降低（ Ｐ＜０． ０５和＜０． ０１）。结论：内毒素在无血清情况下可以和内皮细胞结合，同时可以影响胞内连接蛋白α－Ｃａｔｅｎｉｎ的表达状态，并不依赖是否有血清的存在。

To explore the evidence of the direct combination of endothelial with cultured endothelial cells and the ef- fects of endotoxin on intracellularα-Catenin,α-Catenin so as to study the features of endotoxic damage to endothelial cells. Methods: (1)After the FITC labeled endotoxin(2μg/ml)was co-incubated with cultured endothelial cells in the absence of serum,the changes of F values on endothelial cells was detected by fluorescent spectrophotometer. (2)Cultured endothelial cells without serum were stimulated by cndotoxin (2μg/ml)and the changes of intracellular α-Catenin expression were measured with ELISA. Results: The F values at 10.20.40 and 60 min after that endothellal cells were incubated with endotoxin were 1. 87 (P <0. 05)、 3. 97、 3. 41 (P < 0. 01)and 2. 02(P <0. 01)times of that of control group respecfively. The expression of intracellular α-Catenin was much lower than that of control group (P <0. 05 or P <0. 01)at each designed time points. Conclusions: Endotoxin can combine with endothelial cells in the absence of serum,and interfere with the expressive state of intracellular α-Catenin is independent of serum.