Experiment Study Of The Inhibition Effect And Mechanism Of Danshen Injection Combined With Radiotherapy In Rat's Lewis Pulmonary Tumor Model

Experiment Study of the inhibition effect and mechanism of Danshen injection combined with radiotherapy in Rat's Lewis Pulmonary Tumor ModelObjective:To detective the expression of hypoxia-inducible factor-1α(HIF-1α) and the concentration of vascular endothelial growth factor(VEGF)in blood, observe whether the Danshen injection has the role of reduce the number of hypoxia cells to tumor, improve the state of hypoxia, and enhance the sensitization and raise the efficacy if radiotherapy.Methods:60 healthy female C57BL/6 mice were made into Lewis pulmonary tumor model mice. These mice were randomly divided into 6 groups:model group, Danshen low dose group, Danshen high dose group, radiotherapy group, Danshen low dose+ radiotherapy group, Danshen high dose+ radiotherapy group. Each group's mice accept daily high intraperitoneal injection from the second day,model group's mice received normal saline 0.2ml/one,mice of Danshen low dose group's and Danshen low dose+radiotherapy group's received Danshen injection 20ml/kg/d, mice of Danshen high dose group's and Danshen high dose+ radiotherapy group's received Danshen injection 60ml/kg/d, for 10 days. Mice of radiotherapy group's, Danshen low dose+ radiotherapy group's and Danshen high dose+ radiotherapy group's received 60Coγ2Gy/d×5 days. The routine blood and VEGF expression levels were detected by eye blood.; the quality of the tumors were measured to calculate the inhibition rate; the expression of HIF-1αmRNA in tumor cells were detected by RT-PCR; the microvessel density of tumor,the characteristics of apoptotic cells and the number of organ metastasis were observed on electron microscope.Results:1. Compared with model group, the tumor weight of Danshen low-dose and Danshen high-dose group reduce (P< 0.05), and the inhibition ratio were 21.11%,30.65%. Compared with model group, the tumor weight of radiotherapy group,Danshen low dose+ radiotherapy group and Danshen high dose+ radiotherapy group reduced and the inhibition ratio were 1.01%,36.18%,21.61%. Danshen low dose+ radiotherapy group Compared with radiotherapy group, the inhibition ratio were 35.38%,17.1%. The tumor weight significantly reduced (P<0.01)2. Compared with radiotherapy group, the white blood cell count,red blood cell count of Danshen low dose+ radiotherapy group and Danshen high dose+ radiotherapy group increased (P<0.05),the Platelet count decreased.3. Compareed with model group, the expression of HIF-lamRNA of Danshen low dose group has the trend of decrease, but the expression of Danshen high dose group and radiotherapy group increae. Compareed with radiotherapy group, the expression of HIF-1αmRNA of Danshen low dose+ radiotherapy group and Danshen high dose+ radiotherapy group obviously decreased(P<0.05).This showed that Danshen injection combined with radiotherapy has the role of decrease the expression of HIF-1αmRNA.4. Compared with model group, the concentration of VEGF in tumor of Danshen low dose group,Danshen high dose group,radiotherapy group,Danshen low dose+ radiotherapy group and Danshen high dose+ radiotherapy group significantly reduced (P＜0.05). Compared with radiotherapy group, the concentration of VEGF in tumor and the index of microvessel density of Danshen low dose+ radiotherapy group and Danshen high dose+ radiotherapy group has the trend of decrease.5. Compared with model group, the index of microvessel density of Danshen low dose group,Danshen high dose group,radiotherapy group,Danshen low dose+ radiotherapy group and Danshen high dose+ radiotherapy group significantly reduced (P<0.05).Compared with radiotherapy group, the index of microvessel density of Danshen low dose+ radiotherapy group and Danshen high dose+ radiotherapy group has the trend of decrease (P＜0.01).6. Compared with model group, the tumor cell apoptosis index of Danshen high dose group,radiotherapy group,Danshen low dose+ radiotherapy group and Danshen high dose+ radiotherapy group increased (P<0.05). Compared with radiotherapy group, the tumor cell apoptosis index of Danshen low dose+ radiotherapy group and Danshen high dose+ radiotherapy group increased (P<0.05).This point out Danshen injection has the effection of Induced apoptosis. 7. After the end of treatment, tow lung metastases were found Under light microscopy in Danshen high-dose group,Danshen low dose+radiotherapy group and Danshen high dose+radiotherapy group. While five lung metastases were found in radiotherapy group.Conclusions:Danshen injection combined with radiotherapy could reduce the expression of HIF-1αmRNA and the concentration of VEGF, to increase oxygen pressur of hypoxia cells and inhibit tumor angiogenesis, increase the sensitivity of tumors to radiation, increasing the inhibition ratio and the tumor cell apoptosis index.