Effect of Huoxue Yiqi recipe on proliferation,migration and tube formation of miR-484 overexpressed umbilical vein endothelial cells

Objective:To explore the possible mechanism of Huoxue Yiqi recipe promoting angiogenesis by targeting VEGF with miR-484.Methods:The model of miR-484 overexpressed in HUVEC was prepared and intervened with Huoxue Yiqi Recipe Freeze-dried powder.The effects of Huoxue Yiqi Recipe on proliferation,migration and tubular forming ability of HUVEC were separatly detected by CCK-8 method,cell scratch test and Matrigel three-dimensional forming test in vitro.The effect of Huoxue Yiqi Recipe on VEGFA mRNA expression was detected by real-time fluorescence quantitative PCR.Results:Compared with the control group,the proliferation and migration area of cells in miR-484 overexpression model group decreased,the length,number of branches and nodes involved in tube formation decreased,the tube formation ability decreased significantly,and the expression level of VEGFA mRNA decreased,with statistical significance.Compared with the model group,the cell proliferation ability,migration area,length,number of branches and nodes involved in tube formation,tube formation ability and expression level of VEGFA mRNA in the Huoxue Yiqi Prescription group were up-regulated,and the difference was statistically significant(P<0.05).Conclusion:MiR-484 inhibits the expression of VEGFA and the proliferation,migration and formation of HUVEC,and Huoxue Yiqi recipe has a certain negative regulation effect on this process,which may be a new target of Huoxue Yiqi recipe to promote angiogenesis.

Effect of Bushen Yiqi Huoxue Recipe on Placental Vasculature in Pregnant Rats with Fetal Growth Restriction Induced by Passive Smoking

Interactions of vascular endothelial growth factor （VEGF） with receptors VEGFR1/Fltl and VEGFR2/Flk1, and those of angiopoietins （Ang-1, Ang-2） with receptor Tie2 play important roles in placental angiogenesis. This study investigated vascular morphology and expression of these angiogenic factors in rat placenta on the day 15, 18, 21 of gestation （D 15, D 18 and D21）. The rats were randomly assigned into 3 groups： normal group, model group [fetal growth restriction （FGR） model], and Bushen Tqi Huoxue （BYHR） recipe treatment group （BYHR group, the pregnant rats with FGR were treated with BYHR recipe）. Morphological analysis indicated that during initial villous formation, fetal nucle- ated erythrocytes （FNEs） appeared in maternal blood sinus （MBS）. Subsequently, FNEs were sur- rounded by endothelial cells to form fetal capillary （FC） and then by trophoblast cells to form villi. As pregnancy proceeded, FC density increased progressively with increasing endothelial identification staining （EIS） in normal and BYHR groups. Whereas, villous formation was suppressed, normal in- crease in FC density was impaired and EIS was weakened in model group. Quantitative PCR analysis showed that VEGF and Flkl mRNA increased over gestation in all groups, indicating that VEGF might play a pivotal role in FC growth during late gestation. VEGF mRNA was increased on D15, while de- creased on D21 in model group as compared with normal group and BYHR group. Immunohistochemi- cally, Ang-2 protein was highly expressed in FNEs, gradually disappeared as villi matured, and decreased over gestation in all groups, indicating that Ang-2 might play a pivotal role in villous formation, which was further supported by decreased Ang-2 mRNA and protein expression in model group on D 15. Ang-1 mRNA, Tie2 mRNA and Ang-1/Ang-2 ratio increased from D15 to D18 in all groups as placenta matured. Ang-1 mRNA, Tie2 mRNA and Ang-1/Ang-2 ratio were decreased on D18 in model group as compared with normal and BYHR groups, indicating delayed maturity of FGR placenta. Alterations in angiogenic factors may result in altered placental vasculature and cause placental insufficiency. BYHR recipe could balance the angiogenic factors to promote the formation and maturation of FGR placental vasculature.

Effects of Yiqi Huoxue Recipe （益气活血方） and Coxsackie Virus B Type 3 on the Expression of Ribosomal Protein S20 in Rat Cardiac Myocytes

Objective： To study the effects of Yiqi Huoxue Recipe （益气活血方） and Coxsackie B virus type 3 （CVB3） on the expression of ribosomal protein S20 in rat cardiac myocytes, to explore the pathogenesis of myocarditis induced by CVB3 and the mechanism of Yiqi Huoxue Recipe on gene level, and to further investigate whether Yiqi Huoxue Recipe is an effective prescription for CVB3 myocarditis. Methods： A modified suppression subtractive hybridization （SSH） was used to isolate differentially expressed genes between the CVB3 infection group and the treatment group with Yiqi Huoxue Recipe. The results were further verified by fluorescence RT-PCR. Results： The results of SSH showed that the gene expression of ribosomal protein S20 in the treatment group was higher than that in the CVB3 infection group （P〈0.05）, which agreed with the results of fluorescent RT-PCR. Conclusion： Down-regulation of ribosomal protein S20 mRNA expression might be one of the mechanisms in CVB3 myocarditis, and Yiqi Huoxue Recipe could achieve the treatment of viral myocarditis by regulating the expression of ribosomal protein S20.

Clinical Observation on Effect of Yiqi Huoxue Recipe (益气活血方) in Comprehensive Treatment on Advanced Stage of Esophageal Cancer

Objective: To observe the effect of Yiqi Huoxue (replenishing Qi and activating blood circulation, YQHX) recipe in patients with advanced stage of esophageal cancer treated with radiotherapy and intervention chemotherapy. Methods:In comparing 31 patients who were treated with radiochemotherapy (RCT, Group A), and 31 patients treated with RCT plus YQHX recipe(Group B). Results:Immediate effective rates were 48.4% in Group A and 64.5% in Group B (P>0.05). The function of bone marrow in Group B was obviously better than that in Group A. The 1 year metastasis rate in Group A was higher than that in Group B, but the longterm survival rate of Group B was obviously higher than that in Group A (P<0.05). Conclusion: YQHX recipe could reduce the bone marrow suppression caused by RCT, lower the metastasis rate, prolong the longterm survival and improve the quality of life.

Clinical and Experimental Study on Prevention and Treatment of Acute Radiation Injury in Nasopharyngeal Carcinoma Patients by Yiqi Huoxue Yangyin Recipe (益气活血养阴方)

Objective: To observe the effect of Chinese herbal medicine in alleviating acute toxic adverse effect of radiotherapy and on growth of stromal cells in bone marrow. Methods: Seventy two patients with nasopharyngeal carcinoma (NPC) were randomly divided into two groups. Radical radiotherapy was applied to both groups. In the radiotherapeutic period, to the 36 patients in the treated group, Yiqi Huoxue Yangyin Recipe (YHYR) was given additionally by oral taking and compressing on radiation area, and to the 36 patients in the control group, vitamin B 12 solution was given for gargling or compressing. Experimental study was carried out in rats of two groups, irradiated with 5.0 Gy X ray and treated with YHYR or normal saline intraperitoneally, to observe the colony forming unit fibroblastoid (CFU F) in cultured bone marrow stromal cells (BMSC), taken from the rats at different time.Results: The oral mucomembranous radiation damage occurred in 47.2% of the treated group when the dose of radiation reached to 41.4±9.4 Gy, while in the control group, it was 91.7% when the dose reached to 30.9±8.9 Gy. The skin radiation damage occurrence rate in the two groups was 13.9% and 33.3% respectively when the dose of radiation reached to 50.2±5.6 Gy and 43.2±6.3 Gy respectively. Comparison of the two groups showed significant difference ( P <0.01), the radiation damage was significantly slighter in the treated group. Experimental study showed that the rats in the two groups were significantly different in occurrence and degree of bone marrow function inhibition, P <0.01. The numbers of CFU F in cultured BMSC of radiation damaged rats taken at respective different culture time were also significantly different between the two groups, P < 0.01 .Conclusion: YHYR could significantly reduce acute radiation damage of mucomembrane and skin, alleviate the inhibition on bone marrow function, and eliminate the injury of radiation on BMSC.