ABSTRACT
ObjectiveTo study the effect of Bushen Huoxuetang on the apoptosis and the expression of B-cell lymphoma （Bcl-2）-associated X protein （Bax）/ Bcl-2 and cleaved cysteine-containing aspartate proteolytic enzyme-3 （cleaved Caspase-3） in the nude mouse model of bone metastasis of breast cancer， and explore the mechanism of Bushen Huoxuetang in inhibiting bone destruction. MethodThirty BALB/c female nude mice were randomly assigned into blank group （n=6） and model group （n=24）. The suspension of 4T1 breast cancer cells was injected into the tibia of mouse right lower limb to establish model of bone metastasis of breast cancer. The successfully modeled nude mice were randomly assigned into model group， Bushen Huoxuetang group， zoledronic acid group， and combined drug group， with 6 mice in each group. Bushen Huoxuetang was administrated at a dose of 36.67 g·kg-1， once a day， and zoledronic acid was administrated by subcutaneous injection at a dose of 100 μg·kg-1， twice a week. The combined drug group was administrated with the same doses of Bushen Huoxuetang group by gavage and zoledronic acid by subcutaneous injection. The mice in the blank group and the model group were administrated with the same volume of distilled water by gavage for 14 days. On the next day at the end of drug administration， the mice were sacrificed by cervical dislocation. The general situation and weight changes of the mice were examined. The right lower limb was collected， and X-ray scanning and hematoxylin-eosin （HE） staining methods were used for observation of pathological changes in the bone. The terminal deoxynucleotidyl transferase dUTP nick-end labeling （TUNEL） was employed to detect the apoptosis of bone tissue in nude mice， and Western blot to determine the expression of Bax/Bcl-2 and cleaved Caspase-3 in the bone tissue. ResultCompared with the blank group， the modeling reduced the body weight （P<0.01） and increased the right lower limb weight of the nude mice （P<0.01）. Compared with the model group， Bushen Huoxuetang， zoledronic acid， and their combination increased the body weight （P<0.01） and decreased the right lower limb weight （P<0.01）. Compared with the blank group， the other groups showed obvious tumor cell atypia， deep nuclear staining， and clear bone metastasis， and the model group showed obvious osteolytic damage in right lower limb and loss of proximal tibia and knee joint. Compared with the model group， Bushen Huoxuetang， zoledronic acid， and their combination reduced the osteolytic lesions in the right lower limb and recovered part of the bone structure， demonstrating an inhibitory effect on bone destruction. The TUNEL assay showed that the model group had lower apoptosis rate of bone metastatic tumor cells than the blank group， Bushen Huoxuetang group， zoledronic acid group， and combined drug group （P<0.01）. Compared with the blank group， the modeling down-regulated the expression of Bax and cleaved Caspase-3 （P<0.01） and up-regulated the expression of Bcl-2 （P<0.01）. Compared with the model group， Bushen Huoxuetang， zoledronic acid， and their combination up-regulated the expression of Bax （P<0.01） and cleaved Caspase-3 （P<0.05， P<0.01） and down-regulated the expression of Bcl-2 （P<0.05， P<0.01）. ConclusionBushen Huoxuetang may inhibit bone destruction in the nude mouse model of bone metastasis of breast cancer by up-regulating the expression of Bax， down-regulating the expression of Bcl-2， activating cleaved Caspase-3， and further inducing apoptosis.
ABSTRACT
OBJECTIVE:To investigate the utilization and trend of bisphosphonates drugs in patients with bone metastasis of breast cancer from Zhejiang province,so as to provide reference for the further promotion of their rational use. METHODS:The utilization of this type of drugs in 11 hospitals of Zhejiang province during 2010-2015 was analyzed statistically,counted and ranked by using the method of cost ranking analysis and DDDs analysis in the respect of amount,DDDs,DDC,etc. RESULTS:The amount of bisphosphonates drugs was increased steadily in 6 years(increasing greatly in 2011),increased by 96.26% in 2015,compared to 2010. Top 3 drugs in the list of amount were Zoledronic acid for injection(imported),Zoledronic acid for injection(domestic)and Pamidronate disodium for injection. Among them,Zoledronic acid for injection(imported)basically showed an upward trend,while Pamidronate disodium for injection showed decline trend. Top 3 drugs in the list of DDDs were Zoledronic acid for injection(domestic),Zoledronic acid for injection(imported)and Pamidronate disodium for injection;DDDs of Zoledronic acid for injection(domestic)and Zoledronic acid for injection(imported)were generally on the rise,while that of Pamidronate disodium for injection showed an overall downward trend. B/A value of bisphosphonates ranged 0.33-2.00,among which B/A value of Zoledronic acid for injection(domestic)was 2.00 and that of Zoledronic acid for injection(imported)was 0.50 during 2012-2015. The first and second drugs in the list of DDC were Clodronate disodium for injection and Zoledronic acid for injection(imported). CONCLUSIONS:The total amount of bisphosphonates drugs in patients with bone metastasis of breast cancer from 11 hospitals of Zhejiang province during 2010-2015 was increased steadily. Some varieties of second and third generation bisphosphonates with definite curative effect,good safety and reasonable price are dominant in clinical use. The selection and use of bisphosphonates is basically reasonable,but price is still the primary factor restricting the further wide application of some varieties.
ABSTRACT
AIM To observe the drug pair Baizhu-Fuzi's protection on the breast cancer nude mice with bony metastasis and to explore the mechanism of bone metastasis.METHODS Nude mouse models of breast cancer with bone metastasis were developed through injection of breast cancer cell MDA-MB-231BO into the left ventricle.Nude mice were randomly divided into group A and group B.Group A were subdivided into model group,zoledronic acid group and the drug pair of Baizhu-Fuzi group;and Group B were subdivided into sham-operation group,model group,zoledronic acid group and Baizhu-Fuzi group were subsequently administered with the intervention accordingly.The nude mice in group A had their surviving time and the weight changes observed;and those in group B had the degree of bone metastasis examined.Tartrate resistant acid phosphatase (TRAP) method for quantitative determination of osteoclast in bone metastasis,and ELISA method were employed to check the content of TGF-β1 and PTHrP in serum.RESULTS Compared to the model group,Baizhu-Fuzi group displayed distinctly longer survival time (P ＜ 0.05),reduced rate of weight loss 6 weeks after modeling (P ＜ 0.05),significantly declined degree of bone metastasis (P ＜ 0.01),and significantly decreased quantity of TRAP (+) cell (P ＜ 0.05) and serum TGF-β1 and PTHrP (P ＜ 0.05).CONCLUSION The drug pair,Baizhu-Fuzi's influence in TGFβ signal path control and PTHrP expression reduction may contribute to the weight loss management,prolonged survival time,osteolytic bone defect rectification in mouse models of breast cancer with bone metastasis.