Clinical Study And Experiment For The Mechanism Of Promoting Angiogenesis Of Shengmai Chenggu Capsule

ObjectiveTo study the angiogenesis mechanism of Shengmai Chenggu Capsule (SMCGC) and its effects on the function of vasoactive-substance-secreting of endothelial cell, so as to provide experimental evidence for its clinical application to treat avascular necrosis of femoral head and improve the blood supply of femoral head. Further more, to precisely measure the grey value of blood vessels in and around the necrotic femoral heads before and after the treatment of in taking SMCGC and radiological interventional treatment with the technique of digital substraction angiography.To elucidate the basic theory "to promote generating blood vessels by stimulating the blood circulating" of Traditional Chinese Medicine from the viewpoint of modern pharmacology.MethodOn the basis of reviewing the experimental progress and clinical application of therapeutic angiogenesis, combined with the previous results from the same project sponsored by National Scientific Fund Committee (No.30171143) and Scientific Fund of Guangdong Province (No.010251) respectively, this doctoral program was divided into two parts: basic experiment and clinical study. In the first part, MTT method was used to study the influence of SMCGC-containing rabbits' serum on the proliferation of ECV304 cell. After ECV304 cell being stimulated by SMCGC-containing rabbits' serum for 1 hour and 2 hours, ELISA method was applied to measure the VEGF concentration in the cultured liquid;With the same method, the concentration of ET-1, PGI₂ in the culturedliquid was detected;The concentration of NO in the cultured liquid was detected with enzymic method. The micro-structure of ECV304 cell was observed by transmission electronic microscope technique after being treated for 12 hours. In the clinical trial part, patients involved in the study, who suffer from avascular necrosis of femoral head, were divided into two groups, the patients in group one received interventional radiology treatment only, the patients in the other group got interventional radiology treatment as well as SMCGC-intaking, digital subtractive angiography technique was used to observe the vessels in and around the necrotic femoral head before the treatment and 2 months after the treatment, with image-analyzing software the grey value of angiography before and after the treatment was measured, the differences between the relevant were analyzed statistically with t test.ResultIn the experimental part, after ECV304 cell being treated with SMCGC-containing rabbits' serum for 1 hour, the concentration VEGF of the cultured liquid in low-dose-group increased significantly VEGF(P<0.01), two hours later, the VEGF concentration of the cultured liquid in low-dose-group and medium-dose-group increased significantly (P<0.01). After being stimulated with different doses of SMCGC-containing rabbits' serum for 12 hours and 36 hours, the proliferation of ECV304 cell in all groups was not significantly increased(P<0.05), further more, the proliferation of ECV304 cell in medium-dose-rabbit-serum group and high-dose-rabbit-serum group was inhibited significantly after being treated 36 hours(P<0.01). After ECV304 cell being treated with SMCGC-containing rabbits' serum for 1 hour, the concentration of ET-1 in cultured liquid of high-dose-group significantly decreased(P<0.05) ? the concentration of PGI2 in cultured liquid of low-dose-group, medium-dose-group and high-dose-group was significantly decreased(P<0.05), the concentration of NO in all three groups increased (P>0.05);2 hours later, the concentration of ET-1 in cultured liquid of high-dose-group significantly increased (P<0.05), compared with the control group, the concentration of PGI2 in all three groups was still at low level (P<0.05), but the concentration of PGI2 in thecultured liquid of low-dose-group increased significantly compared with that of medium-dose-group and high-dose-group (P<0.05), the concentration of NO in the cultured liquid of all three groups increased (P>0.05). Compared with blank group, medium-dose-group and high-dose-group, the number of chondriosomes in low-dose-group and Ligustrazine group increased , and the size of chondriosomes in Ligustrazine group became larger, there were more and larger transparent bubbles in the ECV304 cells of high-dose-group than in those of other groups.The clinical study results were as follows: two months after the interventional radiological therapy combined with SMCGC-intaking, the grey value of blood vessels in and around the femoral head increased significantly (P<0.05), the hip pain was relieved and the Harris scores increased significantlyoConclusionThe results of the basic experiment indicated that low dose SMCGC-containing rabbits' serum could stimulate ECV304 cell to secret VEGF significantly after being stimulated 1 hour, both low dose and medium dose SMCGC-containing rabbits' serum could stimulate ECV304 cell to secret VEGF significantly after being stimulated hours, however the SMCGC-containing rabbits' serum of the high-dose-group had no such effect, which elucidates stimulating the endothelial cell to secret VEGF may be the mechanism of promoting angiogenesis of SMCGC, but the effect did not increase with the dose. SMCGC-containing rabbits' serum of all the three groups inhibited the PGh-secreting function of ECV304 cell could explained the mechanism of its anti-inflammation and pain-killing effect;SMCGC-containing rabbits' serum of all the three groups promote ECV304 cell to secret NO, as to the ET-1-secreting function, high dose SMCGC-containing rabbits' serum had a two-direction effect, inhibiting first and promoting later. SMCGC-containing rabbits' serum had no proliferation-promoting effect for the ECV304 cell cultured in vitro and had no obvious influence on the microstructure of the ECV304 cell, while Ligustrazine might do some damage to the chondriosomes of ECV304 cells. The results of clinical study showed that interventional radiological therapy combined with SMCGC-intaking had a better therapeuticangiogenesis effect than merely interventional radiological therapy, further more, interventional radiological therapy combined with SMCGC-intaking had a better therapeutic outcoming when it came to symptoms-relieving and function-improving. All in all, the conclusion can be drawn that SMCGC stimulates the angiogenesis by promoting the endothelial cell to secret VEGF, it also had a profound influence on the secreting function of other vascular substances such as ET-1, NO and PGh, these results provided a reliable experimental and clinical background to use SMCGC as one effective method to treat avascular necrosis of femoral head.