Hydroxy Safflower Yellow A Study Of The Regulation Mechanism Of Vascular Endothelial Cells

The growth of tumor needs blood vessels supplying nutrition and removing metabolite. How to intercept the form of tumor blood vessels has becoming an important topic in the study of tumor. Studies indicated that there are at least 30 growth factors having relation with the growth of tumor blood vessels, among which the most direct vasotropic factors are VEGF and bFGF.The bolting of medicines that inhibit the growth of tumor vessels depends on inter-experiments and extra-experiments, in inter-experiments the CAM is used most, which is an excellent mold to bolt a large of blood inhibited medicines; MTT is generally chosen in extra-molds, which is used to detect the medicines that inhibit the growth of VEC. In the course of the forming of new vessels, the generation of VEC is the most elemental and important component, so the forming of new vessels can be inhibited by inhibiting the growth of VEC. Because the depending of TC(tumor cell) and VEC each other, which mediates the forming of tumor vessels, the study of the form of extro-tumor vessels needs that VEC should be growth in the circumstance of cultivating TC and VEC together. The theory of Chinese Medicine considers that 'stagnancy of qi and blood stasis'is one of the most important courses lead tumor forming, 'blood stasis' is the basic symptom of tumor, for this reason, the method of promoting blood circulation by removing blood stasis has an important position in the study of resisting tumor. Honghua is a traditional herb to promote blood circulation and remove blood stasis, which is used to treat many kinds of tumor such as CACX, but it's mechanism of action is seldom reported. HSYA is the essential component of Honghua, at present, there is few reports about HSYA. Now, there is not report about the action of inhibiting new vessels forming of HSYA.By preliminary test, we have bolted HSYA from 10 herbs or active components of promoting blood circulation and removing blood stasis, which can inhibit the form of CAM new vessels and the generation of the VEC of human umbilical vein. To approach the action and mechanism to TC, we add the supernatant of TC to VEC and construct an abnormal mold of VEC generation, by the experiments of MTT, PCR and ELISA we observed the action and mechanism of HSYA to normal and abnormal generated VEC, approached the potential action mechanism of inhibiting new vessels in the layer of gene expression and proteinum expression.This study consists 3 partsPart I: the research on the mechanism of affecting the growth of CAM vesselsBecause neoformative vessels is most related with bFGF and VEGF, this study designed bFGF, VEGF and the primer of it's receptor, by the experiment of the generation of CAM vessels, draw the materials from CAM tissues consist account of vessels, by the experiment of RT-PCR, reserve the influence of HSYA on the expression of bFGF, VEGF, VEGF-R(flt-1) in CAM tissues. The consequence is: HSYA can depress the expression of bFGF, VEGF, VEGF-R(fit-l) in CAM vessels remarkably, especially the expression on bFGF mRNA, there is a very remarkable difference compared with the normal group.Part II, the action on ECV304 cultivated in the circumstance of physiology and pathologyThe form of tumor vessels is begin with the abnormal generation of CAM, so the form of the new vessels can be inhibited by inhibiting the generation of CAM. By the experiment of MTT, we observed the generated action on ECV304 of the different concentrate of HSYA, the result is: low concentrate of HSYA(5.35mg/L-0.16mg/L) can inhibit the growth of ECV304, the inhibit action is increased by the decrease of concentrate, the action is most powerful when the concentrate is between0.66mg/L and 0.16mg/L, which is remarkable(p<0.001).To approach the action of HSYA on the tumor VEC, we added TC supernatant into VEC and erect a mold of abnormal generated blood VEC, by the experiments of MTT observe the effect of HSYA. The consequence is: the growth of VEC is rapid in the group without medicine, compared with the contrast group the effect of promotin...