Tcm Injections, The Regulation Of Tumor Angiogenesis, The Mechanism Of Signal Transduction Pathways

Background: Traditional Chinese medicine in prevention and treatment of tumor have a long history. The TCM's treatment is an important part of combined therapy of tumor. Anti-angiogenic provide new ideas for the study of the mechanism of Chinese medicine. In the 70s of the 20~th century, Folkman first discovered that the growth of tumor depends on the genesis of the newly-born blood vessels. The tumor can't grow beyond 2-3 mm3 for lack of oxygen and nutrient without angiogenesis. With angiogenesis the tumor can obtain nutrition and the capillaries can provide a chance for tumor metastasis. In last years, research on angiogenic inhibition of tumor is gaining increasing attentioa Angiogensis is a complex, involves the following steps: dissolve the vascular endothelial basal membranl, endothelium cell migrate to tumor tissue, endothelium cell proliferate, form new capillary, form new basal membrane. The proliferation of the vascular endothelial cell play an important role in angiogenesis. The proliferation of the vascular endothelial cell is regulated by VEGF/VEGFR, PKC-MAPK pathway. In last years, research on angiogenic inhibition of tumor is gaining increasing attentioaObject: To investigate the effect to KLT and HCS on angiogenesis, and the mechanism of which is regulation in VEGF/VEGFR, PKC-MAPK pathway.Content: this study consists of four parts. The main content is as follows:1. Anti-angiogenic effects of KLT and HCS in vivo studyIn this study, we aim to investigate the effect to KLT and HCS on angiogenesis, and the ralationship of which with the growth and metastasis of tumor.Mice were randomly divided into 8 groups, KLT group, HCS group, and Rg3 group (positive control), normal saline group(negative control). All were transplanted with Lewis lung cells. The tumor inhibition rate, the number of tumor nodule in the lung and the quantitation of microvessel density were observed 2 week later.The results showed that KLT and HCS effectived reduced the quantitation of microvessel density in tumor. Compare with control group, the quantitation of microvessel density were statistically decreased (P<0.05).KLT and HCS effective reduced the number of tumor nodule in the lung. The inhibited rates of KLT and HCS were 53-82% and 45-22%,respectively. Compared with the control group, the tumor inhitioiy rate in KLT group and HCS group were statistically decreased (P<0.05).The growth of tumors were inhibited in mice with KLT and HCS inhibited the growth of tumors ,to certain extent .The tumor inhibited rates of KLT and HCS were 19-31% and 2140%, respectively. Compared with the control group, the tumor inhitiory rate in KLT group and HCS group were statistically decreased (P<0.05).This result hints that KLT and HCS could inhibit angiogenesis dramatically. The growth and metastasis of tumor, through direct or indirect route, were decreased by inhibiting angiogenesis. 2.Anti-angiogenic effects of KLT and HCS in vitro studyIn this study, we aim to observe the effect of KLT on HUVECs proliferation in various conditionsHUVECs was cultured and induced by VEGF. The cell proliferation was tested by methyl thiazolyl tetrazolium (MTT) methodThe results showed that KLT and HCS could obviously inhibit the proliferation of HUVECs induced by VEGF with IC50 values of 1 Oul/ml and 0.1 ug/ml, respectively. Compared with the control group, the difference was significant (PO.05). But KLT and HCS showed no direct effect on growth of HUVECs (P>0.05).This result hints that the mechanism of anti-angiogenic of KLT and HCS might be the inhibition on proliferation of HUVECs. KLT and HCS could inhibit the VEGF induced HUVECs proliferation, but them had not effect on directly inhibiting proliferation of HUVECs in normal conditioa The results showed that a bi-directional regulation on inhibiting proliferation of HUVECs was shown in KLT and HCS. 3.To study the mechanism of KLT and HCS in regulating VEGF/VEGFR, PKC-MAPK signal transductictionIn this study, we aim to explore the mechanism of KLT and HCS in inhibiting the proliferation of HUVECs through regulating VEGF/VEGFR, PKC-MAPK pathway.The expression of VEGF of implanted tumor of mice was tested by immunohisochemistry. The cultured HUVECs were randomly divided into 5 groups, the normal group, the control group, KLT group, HCS group, Rg3 group. Normal saline was given to the normal group and the control group,10 ul/ml KLT, O.lug/ml HCS and lOOug/ml Rg3 was given to KLT group, HCS group, Rg3 group respectively. Except for the normal group, the other 4 groups were induced by 5u]/ml VEGF for 48 hours. The expressions of KDR, PKC and ERK1/2 were observed from the cultured HUVECs by western blotting methodThe results showed that KLT and HCS lowered the expression of VEGF of implanted tumor of mice. Compare with control group, the expression of VEGF were statistically decreased/The expressions of KDR, PKC and ERK1/2 in KLT group and HCS group were all lower than those in the...