Effect of Bushen Huoxuetang on Expression of Bax/Bcl-2 and Cleaved Caspase-3 in Nude Mice with Bone Metastasis of Breast Cancer

您当前的位置：

首页 >

文章列表页 >

Effect of Bushen Huoxuetang on Expression of Bax/Bcl-2 and Cleaved Caspase-3 in Nude Mice with Bone Metastasis of Breast Cancer

更新时间：2023-05-17

|

- Effect of Bushen Huoxuetang on Expression of Bax/Bcl-2 and Cleaved Caspase-3 in Nude Mice with Bone Metastasis of Breast Cancer
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 8, Pages: 133-141(2023)
- 作者机构：
  
  1.益阳市第一中医医院，湖南益阳 413000
  2.湖南中医药大学，长沙 410208
  3.湖南中医药大学第一附属医院，长沙 410021
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20222426
  CLC： R22;R242;R2-031;R285.5
- Published：20 April 2023，
  
  Published Online：25 October 2022，
  
  Received：04 May 2022，
- 稿件说明：
扫描看全文
朱蓉,郭雪斌,杨满英等.补肾活血汤对乳腺癌骨转移模型裸鼠Bax/Bcl-2及cleaved Caspase-3蛋白表达的影响[J].中国实验方剂学杂志,2023,29(08):133-141.

ZHU Rong,GUO Xuebin,YANG Manying,et al.Effect of Bushen Huoxuetang on Expression of Bax/Bcl-2 and Cleaved Caspase-3 in Nude Mice with Bone Metastasis of Breast Cancer[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(08):133-141.
朱蓉,郭雪斌,杨满英等.补肾活血汤对乳腺癌骨转移模型裸鼠Bax/Bcl-2及cleaved Caspase-3蛋白表达的影响[J].中国实验方剂学杂志,2023,29(08):133-141. DOI： 10.13422/j.cnki.syfjx.20222426.

ZHU Rong,GUO Xuebin,YANG Manying,et al.Effect of Bushen Huoxuetang on Expression of Bax/Bcl-2 and Cleaved Caspase-3 in Nude Mice with Bone Metastasis of Breast Cancer[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(08):133-141. DOI： 10.13422/j.cnki.syfjx.20222426.

摘要

目的

2

观察补肾活血汤对乳腺癌骨转移模型裸鼠细胞凋亡及B细胞淋巴瘤-2（Bcl-2）相关X蛋白（Bax）/Bcl-2及切割型胱天蛋白酶-3（cleaved Caspase-3）蛋白表达的影响，探讨其抑制骨质破坏的作用机制。

方法

2

将30只BALB/c雌性裸鼠随机分成空白组6只、造模组24只。将乳腺癌细胞株4T1经裸鼠右下肢胫骨注射构建乳腺癌骨转移模型，将成膜裸鼠随机分为模型组、补肾活血汤组、唑来膦酸组、联合用药组，每组6只。补肾活血汤组予以补肾活血汤中药灌胃36.67 g·kg

-1

，每日1次，唑来膦酸组予以皮下注射100 μg·kg

-1

，每周2次，联合用药组给予相同剂量补肾活血汤组灌胃联合唑来膦酸皮下注射，正常组和模型组予以等体积蒸馏水灌胃，连续14 d，次日脊柱脱臼法处死，观察裸鼠一般情况及体质量变化，截取每组裸鼠右下肢，称重，X射线扫描及苏木素-伊红（HE）染色观察裸鼠骨组织形态学变化、原位末端标记法（TUNEL）检测裸鼠骨组织凋亡情况，蛋白免疫印迹法（Western blot）检测裸鼠骨组织中Bax/Bcl-2及cleaved Caspase-3蛋白表达情况。

结果

2

与空白组比较，模型组第5~15天裸鼠体质量显著减轻（

P

<

0.01），右下肢组织质量显著增加（

P

<

0.01），与模型组比较，补肾活血汤组、唑来膦酸组、联合用药组第10~15天裸鼠体质量显著增加（

P

<

0.01），右下肢组织质量显著降低（

P

<

0.01）。HE染色显示与空白组比较，模型组、补肾活血汤组、唑来膦酸组、联合用药组裸鼠肿瘤细胞异型性明显，核染色深，骨转移明确。X射线扫描示与空白组比较，模型组裸鼠右下肢出现明显溶骨性破坏，胫骨近段、膝关节完整结构基本丧失，与模型组比较，补肾活血汤组、唑来膦酸组、联合用药组右下肢溶骨性病变明显减少，部分结构完整，对骨质破坏具有抑制作用。TUNEL染色显示与空白组比较，模型组骨转移瘤细胞凋亡率降低，与模型组比较，补肾活血汤组、唑来膦酸组、联合用药组裸鼠骨转移瘤中细胞凋亡率显著升高（

P

<

0.01）。Western blot显示，与空白组比较，模型组骨转移瘤中Bax、cleaved Caspase-3蛋白表达显著降低（

P

<

0.01），Bcl-2蛋白表达显著升高（

P

<

0.01），与模型组比较，补肾活血汤组、唑来膦酸组、联合用药组骨转移瘤Bax蛋白含量明显升高（

P

<

0.01），cleaved Caspase-3蛋白表达明显升高（

P

<

0.05，

P

<

0.01），Bcl-2蛋白表达明显下调（

P

<

0.05，

P

<

0.01）。

结论

2

补肾活血汤组对乳腺癌骨转移模型裸鼠具有抑制骨质破坏作用，其机制可能和上调Bax，下调Bcl-2，激活cleaved Caspase-3诱导细胞凋亡相关。

Abstract

Objective

2

To 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.

Method

2

Thirty 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.

Result

2

Compared 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）.

Conclusion

2

Bushen 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.

关键词

补肾活血汤乳腺癌骨转移凋亡B细胞淋巴瘤-2（Bcl-2）Bcl-2相关X蛋白（Bax）切割型胱天蛋白酶-3（cleaved Caspase-3）

Keywords

Bushen Huoxuetangbone metastasis of breast cancerapoptosisB-cell lymphoma （Bcl-2）Bcl-2-associated X protein （Bax）cleaved cysteine-containing aspartate proteolytic enzyme-3 （cleaved Caspase-3）

references

SUNG H，FERLAY J，SIEGEL R L，et al.Global cancer statistics 2020：GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries［J］.CA Cancer J Clin，2021，71（3）：209-249.

SOERJOMATARAM I，BRAY F.Planning for tomorrow：Global cancer incidence and the role of prevention 2020-2070［J］.Nat Rev Clin Oncol， 2021，18（10）：663-672.

张宇，解新鹏，李峰，等.唑来膦酸对乳腺癌治疗相关骨流失保护作用的研究进展［J］.中国骨质疏松杂志，2021，27（12）：1853-1858.

毛昀，陈峥，褚雪镭，等.国医大师朱良春治疗骨转移临证经验［J］.湖南中医药大学学报，2020，40（9）：1101-1105.

范洪桥，周亮，刘丽芳，等.谭新华治疗乳腺癌骨转移经验撷菁［J］. 中国中医基础医学杂志，2020，26（8）：1175-1176，1182.

王磊，沙湖，王翠平，等.补肾活血汤治疗膝骨关节炎合并骨质疏松症疗效及对骨代谢标志物水平的影响［J］.中华中医药学刊，2021，39（6）：225-228.

方颖，胡金辉，杨争，等.补肾活血汤联合唑来膦酸治疗乳腺癌骨转移的临床研究［J］.湖南中医药大学学报，2015，35（9）：55-57.

袁博，胡金辉，周亮，等.补肾活血汤对乳腺癌患者芳香化酶抑制剂治疗后骨量减少的影响［J］.中医药导报，2016，22（21）：42-44.

王贤彬，胡金辉，袁博，等.补肾活血汤联合化疗对三阴性乳腺癌骨转移患者免疫功能及生活质量的影响［J］.中医药导报，2018，24（9）：55-57.

王贤彬.补肾活血汤对裸鼠乳腺癌骨转移模型和Jagged_1/Notch及TGF-β通路蛋白的干预研究［D］.长沙：湖南中医药大学，2018.

刘德果.补肾活血汤对裸鼠乳腺癌骨转移瘤及VEGF、bFGF、MVD影响的研究［D］.长沙：湖南中医药大学，2017.

杨争，胡金辉，王希，等.补肾活血汤对SDF-1α诱导的人乳腺癌MDA-MB-231细胞增殖及迁移的影响［J］.时珍国医国药，2016，27（8）：1868-1870.

孟树.补肾活血汤对裸鼠乳腺癌骨转移模型及OPG/RANK/RANKL系统影响的研究［D］.长沙：湖南中医药大学，2019.

SHOJI R，TSUCHIE H，NAGASAWA H，et al. Development of new mouse breast cancer model of local bone metastasis and verification using bisphosphonates［J］.In Vivo，2022，36（2）：667-671.

RUCCI N，MILLIMAGGI D，MARI M，et al.Receptor activator of NF-kappaB ligand enhances breast cancer-induced osteolytic lesions through upregulation of extracellular matrix metalloproteinase inducer/CD147［J］.Cancer Res，2010，70（15）：6150-6160.

乔涵.白花丹素和唑来膦酸联合使用抑制乳腺癌骨转移的诊疗体系研究［D］.上海：上海交通大学，2018.

毛昀.补肾类中药调节乳腺癌骨转移生态位的作用机制研究［D］.北京：北京中医药大学，2021.

WANG W，GORDON J L，PHILBRICK K A，et al.Low calcium diet increases 4T1 mammary tumor carcinoma cell burden and bone pathology in mice［J］.PLoS One，2017，12（7）：e0180886.

闵淑慧，胡依，郭芮绮，等.1990—2019年中国女性乳腺癌疾病负担及变化趋势分析［J］.现代预防医学，2021，48（16）：2941-2945，2956.

PALUCH-SHIMON S，PAGANI O，PARTRIDGE A H，et al.ESO-ESMO 3rd international consensus guidelines for breast cancer in young women （BCY3）［J］.Breast，2017，35：203-217.

WESTPHAL T，GAMPENRIEDER S P，RINNERTHALER G，et al.Cure in metastatic breast cancer［J］.Memo，2018，11（3）：172-179.

郑亚男，李梅，赵基源.乳腺癌骨转移分子机制及靶向药物的研究进展［J］.基础医学与临床，2021，41（3）：433-437.

HIRANO F，OKUMA K F，ZENKE Y，et al.Disturbance of osteonal bone remodeling and high tensile stresses on the lateral cortex in atypical femoral fracture after long-term treatment with Risedronate and Alfacalcidol for osteoporosis［J］.Bone Rep，2021，14：101091.

裴晓华，彭艳梅.中医药治疗恶性肿瘤70年［J］.中国肿瘤外科杂志，2019，11（5）：305-308.

毛昀，李林潞，褚雪镭，等.论乳腺癌骨转移中医病机治法［J］.辽宁中医药大学学报，2021，23（10）：44-47.

MA Y V，LAM C，DALMIA S，et al.Mechanical regulation of breast cancer migration and apoptosis via direct and indirect osteocyte signaling［J］.J Cell Biochem，2018，119（7）：5665-5675.

刘铭，王俊，李丽，等.杜仲绿原酸通过PI3K/Akt/Nrf-2通路增强视网膜母细胞瘤细胞系HXO-Rb44放射敏感性［J］.中国临床解剖学杂志，2019，37（6）：644-649，655.

严斐霞，谢永艳，陈畅，等.熟地黄炮制过程中的化学成分变化和药理作用研究进展［J］.时珍国医国药，2021，32（10）：2493-2495.

郑烨韵，周凤华.红花黄色素抗肿瘤机制新进展［J］.中药药理与临床，2022，doi：10.13412/j.cnki.zyyl.20220608.001http://dx.doi.org/10.13412/j.cnki.zyyl.20220608.001.

GOLDVASER H，AMIR E.Role of Bisphosphonates in breast cancer therapy［J］.Curr Treat Options Oncol， 2019，20（4）：26.

KAYA ÇAKIR H，EROGLU O. In vitro anti-proliferative effect of capecitabine（Xeloda） combined with mocetinostat （MGCD0103） in 4T1 breast cancer cell line by immunoblotting［J］. Iran J Basic Med Sci， 2021，24（11）：1515-1522.

HOU Q，LIN X，LU X，et al.Discovery of novel steroidal-chalcone hybrids with potent and selective activity against triple-negative breast cancer［J］.Bioorg Med Chem， 2020，28（23）：115763.

王坤，钱金权，年建泽，等.放疗加量对T4期乳腺癌改良根治术后患者血清Ki-67、Bax及Bcl-2水平的影响［J］.医学综述，2021，27（11）：2279-2283，2288.

刘斌.Bcl-2、Caspase-3在乳腺癌骨转移患者组织中的表达及相关性研究初探［D］.南充：川北医学院，2017.

肖龙，朱杰，朱丽科，等.基于STAT3通路探讨补肾活血方抑制骨细胞凋亡［J］.中华中医药学刊，2022，40（1）：204-208，后插32-32后插36.

刘永明，张圆，衣鑫，等.高负荷运动抑制鼠软骨细胞自噬并促进其凋亡［J］.细胞与分子免疫学杂志，2021，37（5）：427-434.

0

Views

25

下载量

1

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Related Articles

Effect of Stemona tuberosa Alkaloids on Apoptosis of SMMC-7721 Cells and Expression of Bcl-2， Bax， and Cleaved Caspase-3

Computer-aided Drug Design and Experimental Validation Reveal Molecular Mechanism of Saikosaponin D-induced Apoptosis of Bladder Cancer Cells

Mechanism of Culuan Heji in Regulating AMPK/mTOR/HIF-1/VEGF Pathway to Improve Oxidative Stress and Apoptosis in Rats with Follicular Maldevelopment

Effect of Haizao Yuhutang Combined with Modified Sargassum and Glycyrrhizae Radix et Rhizoma on Expression of p53/p21/Caspase-3 Pathway in Rats with Goiter

Effect of Xiaoyaosan on Apoptosis in Mouse Model of Lipopolysaccharide-induced Depressive-like Behavior in Mice

Related Author

Si LIN

Hua ZHU

Hui-zhen QIN

Xiao-xun WANG

Li-ba XU

Ling-yu DENG

ZUO Ling

LI Yutong

Related Institution

Guangxi University of Chinese Medicine

Guangxi Key Laboratory of Zhuang and Yao Ethnic Medicine， Guangxi University of Chinese Medicine

The Second Affiliated Hospital of Guangdong Medical University

Affiliated Hospital of Guangdong Medical University

Fujian University of Traditional Chinese Medicine（TCM）

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰