Based On TCIPA Pathway,the Intervention Effect And Mechanism Of Tongqiaohuoxue Decoction And Its Removal Formula On Breast Cancer Metastasis Under High Altitude Hypoxia Environment Were Studied

Objective: To study the intervention effect and mechanism of Tongqiaohuoxue Decoction and its removal prescription on tumor metastasis in breast cancer bearing mice under high altitude hypoxia environment.Methods:Seventy-four SPF female Balb/c mice were randomly divided into normal group and model group.After modeling,they were randomly divided into normal group,model group,Tongqiaohuoxuetang group,huoxuetang group and musk group.All the mice were fed in low-pressure oxygen chamber at simulated altitude of5000 m.Drinking water,drinking water,Tongqiao Huoxue Decoction,Huoxue group liquid and musk suspension were given intragastrally at 9:00 a.m.every morning.After 4 weeks of intragastric administration,tumor volume,tumor photon number,tumor weight and tumor inhibition rate were measured.The histopathological changes of tumor in situ were observed by transmission electron microscopy.HE staining was used to observe the number of lung metastases and their proportion in lung tissue area.Platelet count and coagulation function(APTT,TT)were detected.The expression levels of platelet-related factors(CD62P,PDGFR)and angiogenic factors(VEGF,b FGF and HIF-1α)in breast carcinoma in situ were detected by immunohistochemistry.Results:(1)General conditions of mice in each group: compared with the model group,the growth of in situ tumor volume in Tongqiao Huoxue Decoction group and Huoxue Decoction group was slower,the symptom of blood stasis in mice was lighter,the necrosis area in tumor center was smaller,and the feeding condition of mice was better than that in the model group.(2)Tumor volume,tumor photon number,tumor weight and tumor inhibition rate in tumor-bearing mice: the tumor volume in situ was measured and calculated every 3 days after 4 weeks of continuous administration,and the growth curve was plotted according to the tumor volume.The results showed that the tumor volume in the Tongqiao Huoxue Decoction group and the Huoxue Group was smaller than that in the model group.The results showed that compared with the model group,the tumor volume of Tongqiao Huoxue Decoction group,Huoxue group and Shexiang group was smaller,and the differences were statistically significant(P<0.05).It is consistent with the tumor tumor photon number results.Lavage after 4 weeks,executed based in mice,according to take in situ tumor weight,the results showed that: huoxue soup and activating blood group has better effect on tumor suppression,compared with model group,huoxue decoction group and invigorate the tumor weight is small,the difference was statistically significant(P< 0.05),and the inhibitory rate of party: huoxue soup and activating blood group were more than 30%.(3)The histopathological changes of tumor in situ were observed under transmission electron microscope.The tumor cells and microvascular endothelial cells in the tumor in situ tissues of each group had different degrees of pathological changes.Most tumor cells in model group were necrotic,with incomplete structure,swollen mitochondria and enlarged rough endoplasmic reticulum.Vascular endothelial nucleus pyknosis,rough endoplasmic reticulum expansion and autophagy;Most tumor cells in Tongqiao Huoxue Decoction group,Huoxue group and Shexiang group had complete morphological structure,slight mitochondrial swelling and slight expansion of endoplasmic reticulum.Swollen mitochondria,mild dilation of rough endoplasmic reticulum and a small amount of autophagy can be seen in vascular endothelial cells.The pathological changes were as follows: model group > Musk group > Huoxue group ≈ Tongqiao Huoxue Decoction group.(4)The number of lung metastasis in tumor-bearing mice was as follows:model group > Shexiang group > Tongqiao Huoxue Decoction group > Huoxue group;Compared with model group,the number of lung metastasis in Tongqiao Huoxue Decoction group and Huoxue group was significantly different(P<0.05);The ratio of the average area of lung metastasis to lung tissue was as follows: model group >Shexiang group > Tongqiao Huoxue Decoction > Huoxue group.Compared with the model group,the proportion of lung metastasis area to lung tissue in Tongqiao Huoxue Decoction group and Huoxue Group was significantly different(P<0.05).There was no significant difference in lung weight among all groups(P>0.05).(5)Coagulation function of experimental mice(APTT and TT): Compared with normal group,APTT of model group and invigorating blood group was shortened,and the difference was statistically significant(P<0.05).Compared with model group,The APTT of Tongqiao Huoxue Decoction group and Shexiang group was prolonged,and the difference was statistically significant(P<0.05).Compared with normal group,TT of model group,Tongqiao Huoxue Decoction group,Huoxue group and Musk group was shortened,and the difference was statistically significant(P<0.05).Compared with model group,TT of huoxue group was prolonged,and the difference was statistically significant(P<0.05).(6)Platelet count of experimental mice in each group: compared with normal group and model group,platelet count of all drug intervention groups had no significant difference(P>0.05);(7)The positive rates of platelet-related factors(PDGFR and CD62P)in tumor in situ tissues:Compared with the model group,the positive rates of PDGFR and CD62 P in tumor in situ tissues of tumor-bearing mice in Tongqiao Huoxue Decoction group and Huoxue group were decreased,and the difference was statistically significant(P<0.05).(8)The positive rates of angiogenesis related factors(VEGF,b FGF,HIF-1α)in tumor tissue in situ in tumor-bearing mice: there was no statistical significance in the positive rates of VEGF and b FGF in tumor tissue in situ in tumor-bearing mice among all groups(P>0.05).However,the positive rate of HIF-1α in the Tongqiao Huoxue Decoction group was lower than that in the model group,and the difference was statistically significant(P<0.05).Compared with the model group,the positive rate of HIF-1α in the Tongqiao Huoxue Decoction group was decreased,and the difference was statistically significant(P<0.05),while the positive rate of HIF-1α in the Huoxue group and the Shexiang group was increased,and the difference of Huoxue group was statistically significant compared with the model group(P<0.05).Conclusion:Tongqiao Huoxue Decoction and its decoction can effectively inhibit the growth and metastasis of breast carcinoma in situ in tumor-bearing mice under high altitude hypoxic environment,among which Tongqiao Huoxue Decoction and Huoxue group have the best inhibitory effect.It was observed by transmission electron microscopy that Tongqiao Huoxue Decoction and its removal formula could improve the pathological changes of tumor cells and microvascular endothelial cells in vivo in tumor-bearing mice,and make tumor cells more complete in structure,less necrosis,less swelling of mitochondria of vascular endothelial cells,and less expansion of rough endoplasmic reticulum.Further study on the tumor suppressive mechanism of Tongqiao Huoxue Decoction and its removal suggested that the mechanism of inhibiting tumor growth and metastasis might be related to improving blood hypercoagulability and down-regulating the expression of platelet-related factors(PDGFR,CD62P)and regulate the expression of hypoxia-inducible factor HIF-1α.