| Lots of epidemiologic data have verified that human cytomegalovirus (HCMV) has an intimate relationship with the initiation and development of atherosclerosis (AS). The results of some experiments suggest that the antigen and DNA sequence of HCMV exist in the AS lesion tissues, especially in the vascular endothelial cells (VECs). From these findings, we can see that HCMV initiates the formation of AS, which may be related with HCMV-caused lesions of VECs. The lesion of structure and function of VECs is regarded as a pivotal initial factor of AS.VECs not only serve as the physiological barrier structure of vascular tunica intima but also possess many secretory functions. Its synthetic substances like prostacyclin, thromboxane and endothelin, and substances like thrombomodulin ADP enzyme and NO in its surface all contribute to the resistance of the accumulation of platelet, thrombus formation, cell adherence and inhibition of srmooth muscle cell proliferation. The lesion of VECs certainly will accelerate thepathological change of vessels, especially the initiation anddevelopment of AS.At present, a lot of investigations of ginkgo flavone glycoside have been conducted, but most of them have focused on the clinical protection and treatment of AS and only a few of them are on the mechanism at cell level.1. Aims(1) To prove at cellular and molecular levels that the HCMV is one of the factors that damages the structure and function of VEC by studying the effects of HCMV infected VEC on VEC proliferation, cell apoptosis, cell cycle and NO secretion.(2) To provide theoretic basis for the clinical application of traditional Chinese medicinal herbs in the protetction of VEC function and in the protection as well as treatment of AS, by investigating the effects of ginkgo flavone glycoside on the proliferation, cell apoptosis, cell cycle and NO secretion of VEC infected by HCMV.2. Methods(1) Cultured cells were divided into four groups: control group; ginkgo flavone glycoside group, with ginkgo flavone glycoside added into cultured human umbilical vascular endothelial cells (HUVEC304); HCMV group, with HCMV added into cultured HUVEC304; and ginkgo flavone glycoside + HCMV group, with ginkgo flavone glycoside added into cultured HUVEC304 infected by HCMV.(2) The morphology of cultured HUVEC304 in vitro and the morphological changes of HUVEC304 infected by HCMV in vitro were observed by inverted microscope and electron microscope, respectively. The effect of ginkgo flavone glycoside on them was also observed.(3) The poliferation of HUVEC304 infected by HCMV in vitro and the cell cycle and apoptosis of HUVEC304 infected by HCMV were examined by MTT method and flow cytometry method,respectively. The effect of ginkgo flavone glycoside on them was also observed.(4) The NO content of HUVEC304 infected by HCMV and the effect of ginkgo flavone glycoside on the level of NO secretion were measured by the Greiss's method to measure. The effect of ginkgo flavone glycoside on it was also observed.3. Results(1) Under the inverted microscope, HUVEC304 was found cobble shaped. Under electron microscope, the characteristic Weibel-palade structure of VECs and differnt shapes of virus in different period were observed.(2) The detection of proliferative level, cell cycle, cell apoptosis and NO secretion level of HUVEC304 infected by HCMV showed that: 1) HCMV accelerated proliferation of HUVEC304. The MTT OD values were 1.09 + 0.08 and 2.08 + 0.09 in control group and HCMV group, respectively, which increased 91 % compared with that of control group (P < 0.05); 2) HCMV reduced the Gl period cells. The cells in GO/G1 period were 74.4 % and 59.3 % in control group and HCMV group, respectively, which reduced 20.3 % compared with that of control group (P < 0.05); 3) HCMV inhibited the apoptosis level of HUVEC304. The apoptosis cells were 8.3 % and 5.8 % in control group and HCMV group, respectively, which decreased 30.1 % compa... |
PDF Full Text Request