Vascular endothelial cell lesion model of blood-stasis syndrome of type 2 diabetes mellitus
10.3321/j.issn:1006-2157.2009.11.006
- VernacularTitle:2型糖尿病血瘀证血管内皮细胞损伤模型的研究
- Author:
Yong-Hong ZHOU
1
;
Li-Guo CHEN
;
Yuan QU
;
Liang YAN
;
Hai-Lan TANG
;
Shao-Bing CHENG
Author Information
1. 暨南大学医学院
- Keywords:
type 2 diabetes mellitus;
blood-stasis syndrome;
vascular endothelial cell lesion;
cell model
- From:
Journal of Beijing University of Traditional Chinese Medicine
2009;32(11):742-746,754,封3
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish the vascular endothelial cell lesion model of blood-stasis syndrome of type 2 diabetes mellitus.Methods ECV-304 cells of logarithmic phase were intervened and divided into the blank control group (group 1), health group (group 2), blood-stasis syndrome of diabetes mellitus group (group 3) and diabetes mellitus group (group 4). The activities of ECV-340 cells were observed by using MTT test. The morphological changes of the cells were observed by inverted phase contrast microscope, scanning electron microscope and transmission electron microscope. The levels of ET and NO were detected by using radioimmunoassay and nitrate reductase method respectively. The levels of endothelial protein C receptor (EPCR), von Willebrand factor (vWF) and thrombonodulin (sTM) were detected by applying double-antibodies-sandwich-ELISA method. The changes of free calcium in the cells were observed by laser scanning confocal microscope (LSCM) taken Fluo-3/AM as fluorescence indicator. The distribution of actin microfilament was observed by using rhodamine phalloidin staining.Results ①The value of optical density (OD) decreased significantly in group 3 than that in group 2 (P<0.001). ②In group 3 ET level was higher than that in group 1 and that in group 4 (P<0.001), NO level was lower than that in group 1 (P<0.001) and had no statistical difference compared with that in group 4 (P>0.05). ③In group 3 EPCR level was higher than that in group 1 and lower than that in group 4(P<0.001), the level of vWF was higher than that in group 1 (P<0.001) and had no statistical difference compared with that in group 4 (P>0.05), and sTM level was higher than that in group 1 (P<0.001) and had no statistical difference compared with that in group 4 (P>0.05). ④In group 3 the distribution of fluorescence showed uneven and fluorescence intensity was higher than that in group 1 and group 4 (P<0.001). ⑤In group 1 and 2 the distribution of microfilament cytoskeleton was regular and clear with normal cell shape, while in group 3 it was broken and irregular, and in group 4 it was broken but regular.Conclusion ECV-304 cells showed various signs of vascular endothelial cell lesion after the interference by the serum from the patient with type 2 diabetes mellitus. The vascular endothelial cell lesion model of blood-stasis syndrome combined disease and syndrome was established initially.
