Study On The Effect Of Bushen Huoxue Decoction On Breast Cancer Bone Metastasis Model And OPG/RANK/RANKL System In Nude Mice

Objective: To establish a bone metastasis model of breast cancer in nude mice,observe and study the effect of Bushen Huoxue decoction on breast cancer bone metastasis mo del and OPG/RANKL/RANK system protein in nude mice.Methods: Fifty female nude mice were stratified according to physical quality and divided into blank group,model group,zoledronic acid group,Bushenhuoxuetang high,medium and low concentration groups,Huoxue group and Bushen group according to the completely randomized method.There are 8 groups,7mice in each group.All groups except blank group were simultaneously constructed to model breast cancer bone metastasis.The specific construction steps are as follows:(0.1ml,1×107/ml)human breast cancer MDA-MB-231 cell suspension was infused into the left ventricle of nude mice to construct a bone metastasis model of breast cancer.After successful modeling,observe nude mouse life activities closely and record changes in the quality of nude mice.On the second day after successful model replication,except for the blank group,the drugs were treated for 6 weeks for each group.The specific drug intervention methods were as follows: the model group was given 0.2 ml 0.9% NS per day;The zoledronic acid group was given intraperitoneal injection once a week(100 ?g/kg);the low-concentration group of Bushen Huoxue Decoction,the concentration group of Bushen Huoxue Decoction,and the high concentration group of Bushen Huoxue Decoction were administered with crude drug,and the concentration was daily.10.5g/kg,21g/kg,42g/kg;Huoxue group(reduced by Bushen Huoxue Decoction)is given daily with crude drug infusion,the concentration is 3.6g/kg;Bushen group(reduced by Bushen Huoxue Decoction)is given daily for medicine.The stomach was administered at a concentration of 17.4 g/kg.On the 40 th day after modeling,nuclide bone ECT scans were performed on each group of nude mice,and the bone metastasis sites of each group were recorded.On the 42 nd day,the nude mice were subjected to abdominal aorta blood collection and then subjected to cervical dislocation.The bone tissues of each nude mice were removed from the sterilized workbench for pathologically confirmed bone metastasis.HE staining was used to observe the bone tissue of each group.The degree of damage was detected by immunohistochemistry to detect the expression of OPG,RANK and RANKL proteins in each bone tissue.Results:1.After injecting human breast cancer MAD-MB-231 cells into the left ventric le of nude mice,a bone metastasis model of breast cancer can be successfully constructed.2.After the model was constructed,the bone metastases in therapy groups were inhibited in varying degrees.3.The physical quality of the nude mice in the model group was significantly lower th an that in the treatment group,and the difference between the groups was statistically s ignificant(P<0.05).4.The expression of OPG in the bone tissue of nude mice in high concentration of Bushen Huoxue group was higher than that of zoledronic acid group.The difference between the two groups was statistically significant(P<0.05).The expression of RANKL in the high concentration of Bushen Huoxue group was significantly lower than that in the other groups.(P<0.05),the difference between the groups was statistically significant;The expression of RANK has little difference in groups.The expression of OPG in the bone tissue of the nude mice in the model group was significantly lower than that in the treatment groups,the difference between the groups was statistically significant(P<0.05);The expression of RANKL protein in the bone tissue of the nude mice in the model group was significantly higher than that in the treatment groups,the difference between the groups was statistically significant(P<0.05).Conclusions:1.After injecting human breast cancer MAD-MB-231 cells into the left ventricle of nude mice,a bone metastasis model of breast cancer can be successfully constructed.2.Bushen Huoxue Decoction can effectively reduce and inhibit the occurrence of bone metastasis.3.Bushen Huoxue Decoction can enhance the body weight of nude mice.4.Bushen Huoxue Decoction may strengthen the expression of OPG and inhibit the expression of RANKL,in which way to interfere with the OPG/RANK/RANKL system,and inhibit the bone metastasis of breast cancer.