甘草素对乳腺癌内分泌治疗后继发性骨质疏松大鼠Wnt/β-catenin信号通路作用的研究  

Effect of Liquiritigenin on Wnt/β-catenin Signaling Pathway in Rats with Osteoporosis Induced by Endocrine Therapy for Breast Cancer

作　　者：邢如温 李双进 XING Ru-wen;LI Shuang-jin(General Surgery Department,The Sixth Hospital of Shijiazhuang(Shijiazhuang Maternity&Child HealthcareHospital),Shijiazhuang Hebei 050011;Department of Orthopaedics,Shijiazhuang Yanzhao Hospital,Shijiazhuang Hebei 050035)

机构地区：[1]石家庄市第六医院(石家庄市妇幼保健院)普外科,河北石家庄050011 [2]石家庄燕赵医院骨外科,河北石家庄050035

出　　处：《世界中西医结合杂志》2023年第8期1566-1574,共9页World Journal of Integrated Traditional and Western Medicine

基　　金：河北省2021年度医学科学研究课题计划(20210949);国家自然科学基金(81802329)。

摘　　要：目的 探讨甘草素(Liquiritigenin,LG)对乳腺癌内分泌治疗后继发性骨质疏松症(Osteoporosis,OP)大鼠Wntβ-catenin信号通路的影响。方法 将84只SPF级4月龄雌性未孕Sprague Dawley(SD)大鼠,按随机数字表法分为假手术组、去势组、模型组、ALN组、LG-H组、LG-M组、LG-L组,每组各12只。建立继发性OP大鼠模型,除假手术组外,其余组均进行双侧卵巢切除术,假手术组开腹剪去适量脂肪组织,逐层缝合,术后1周给予蒸馏水;去势组、模型组、ALN组及LG各组均行双侧卵巢切除术,术后1周模型组大鼠给予来曲唑,ALN组大鼠给予来曲唑+阿仑膦酸钠,LG各组均给予来曲唑+甘草素,其中LG-H组给予甘草素高剂量300 mg/(kg·d),LG-M组给予甘草素中剂量200 mg/(kg·d),LG-L组给予甘草素低剂量100 mg/(kg·d),假手术组和去势组大鼠给予蒸馏水。各组大鼠灌胃4周后腹主动脉取血,使用酶联免疫吸附测定(Enzyme linked immunosorbent assay,ELISA)检测血清雌二醇(Estradiol,E2)、骨碱性磷酸酶(Bone-alkaline phosphatase,BALP)和I型前胶原氨基端原肽(Eukaryotic Procollagen I N-Terminal Propeptide,PINP);处死大鼠后取右侧股骨,使用显微断层扫描(micro-CT)检测骨密度及骨小梁微结构、HE染色观察骨组织形态;取第四腰椎骨(L4),使用MTS Acumen3型生物力学测试系统进行腰椎压缩试验,分析各组大鼠骨生物力学性能;蛋白免疫印迹法(western blot)检测大鼠胫骨组织Wnt/β-catenin信号通路相关蛋白表达。结果 与假手术组比较,去势组、模型组大鼠最大载荷和刚度、BMD、BV/TV、Tb.Th、Tb.N、E2含量明显降低,Tb.Sp、BALP、PINP明显增加,骨小梁纤细且分布稀疏、结构不完整,Wnt、β-catenin、LRP5、p-GSK3β ser9蛋白表达水平均明显降低;与模型组比较,LG-L组、LG-M组、LG-H组、ALN组大鼠最大载荷和刚度、BMD、BV/TV、Tb.Th、Tb.N、E2含量明显增加,Tb.Sp、BALP、PINP明显降低,骨组织形态均有明显改善,Wnt、β-Objective To investigate the effect of liquiritigenin(LG)on the Wnt/β-catenin signaling pathway in rats with secondary osteoporosis(OP)induced by endocrine therapy for breast cancer.Methods Eighty-four 4-month-old female non-pregnant Sprague Dawley(SD)rats were randomly divided into a sham operation group,an ovariectomy(OVX)group,a model group,an alendronate(ALN)group,an LG-H group,an LG-M group,and an LG-L group,with 12 rats in each group.A secondary OP rat model was established.Except for the sham operation group,bilateral ovariectomy was performed on the remaining groups.In the sham operation group,after the appropriate amount of adipose tissue was cut off,the incision was sutured layer by layer,and distilled water was provided one week after surgery.Bilateral oophorectomy was performed in the OVX group,the model group,the ALN group,and LG groups.One week after surgery,the model group received letrozole,the ALN group received letrozole and alendronate sodium,and the LG groups received letrozole and different doses of LG[LG-H:300 mg/(kg·d),LG-M:200 mg/(kg·d),and LG-L:100 mg/(kg·d)].The sham operation and OVX groups received distilled water.After 4 weeks of gavage,blood was collected from the abdominal aorta for enzyme-linked immunosorbent assay(ELISA)to measure serum estradiol(E_2),bone alkaline phosphatase(BALP),and eukaryotic procollagen I N-terminal propeptide(PINP).The rats were sacrificed and the right femurs were collected for micro-computed tomography(micro-CT)analysis of bone density and microstructure,followed by hematoxylin and eosin(HE)staining for bone tissue morphology.The fourth lumbar vertebra(L4)was used for the lumbar spine compression test with the MTS Acumen3 biomechanical testing system to analyze bone biomechanical properties.Western blot was used to detect the expression of Wnt/β-catenin signaling pathway-related proteins in rat tibial tissues.Results Compared with the sham operation group,the OVX and model groups showed significantly decreased maximum load and stiffness,BMD,BV/TV,Tb.

关 键 词：甘草素 乳腺癌 内分泌治疗 继发性骨质疏松症 WNT/Β-CATENIN信号通路 

分 类 号：R285.5[医药卫生—中药学]