摘要
目的复制大鼠骨质疏松症模型,测定骨组织Wnt3a、β-catenin基因、蛋白表达,探讨杜仲健骨方对大鼠骨质疏松症作用机制。方法将大鼠采用去卵巢方式制备骨质疏松症动物模型,随机分为模型组、阿仑膦酸钠组(阳性药组)、假手术组、杜仲健骨方高、中、低剂量共6组,每组10只。灌胃70 d,每天1次,假手术组干预同模型组。采用酶联免疫测试方法(ELISA)检测血清总碱性磷酸酶(ALP)、抗酒石酸酸性磷酸酶(TRAP)、骨钙素(OC)和I型原胶原N-端前肽(PINP),实时荧光定量聚合酶链式反应(RT-PCR)检测骨组织Wnt3a、β-catenin mRNA表达,蛋白免疫印迹法(Western blot)检测骨组织Wnt3a、β-catenin蛋白表达。结果与假手术组比较,模型组骨密度显著降低(P<0.01);与模型组比较,杜仲健骨方剂量组、阳性药组骨密度明显增加(P<0.01,P<0.05);与假手术组比较,模型组血清骨形成指标ALP、TRAP、OC、PINP含量差异有统计学意义(P<0.01);杜仲健骨方各剂量组、阳性药组大鼠血清中ALP、TRAP、OC、PINP含量与模型组比较差异有统计学意义(P<0.05,P<0.01)。与假手术组比较,模型组骨组织中Wnt3a、β-catenin mRNA及蛋白表达水平差异有统计学意义(P<0.01);杜仲健骨方剂量组、阳性药组Wnt3a、β-cateninm RNA及蛋白表达水平与模型组比较差异有统计学意义(P<0.05,P<0.01)。结论杜仲健骨方作用于Wnt/β-catenin信号通路,发挥治疗骨质疏松症的作用。
Objective To explore the mechanism of Duzhong Jiangu Fang on osteoporosis in rats,the osteoporosis model of rats was established,and Wnt3a,β-Catenin gene and protein expression in bone tissue were determined.Methods The osteoporosis rat models were established by ovariectomization and randomly divided into the model group,alendronate sodium group(positive drug group),sham operation group,and high,medium and low doses of Duzhong Jiangu Fang groups.Intragastric administration was performed once a day for 70 days.ELISA was used to detect serum total alkaline phosphatase(ALP),tartrate-resistant acid phosphatase(TRAP),osteocalcin(OC),and type I procollagen N-terminal propeptide(PINP).The real-time quantitative PCR(RT-PCR)and western blot were used to detect the expression of Wnt3a andβ-catenin mRNA and protein in bone tissue,respectively.Results Compared with the sham group,the bone density was significantly reduced in the model group(P<0.01).Compared with the model group,the bone density was significantly increased in the Duzhong Jiangu Fang and positive drug groups(P<0.01,P<0.05).There was significantly different about the serum bone formation indicators ALP,TRAP,OC and PINP between the sham group and the model group(P<0.01).There was significantly different about the serum ALP,TRAP,OC and PINP between the model group and the Duzhong Jiangu Fang and positive drug groups(P<0.05,P<0.01).There was significantly different about the expression of Wnt3a andβ-catenin between the model group and the Duzhong Jiangu Fang and positive drug groups(P<0.05,P<0.01).Conclusion Du Zhongjian Gu Fang acts on the Wnt/β-catenin signaling pathway and plays a role in the treatment of osteoporosis.
作者
邢威
陈雁雁
王树人
王艳丽
王文姌
张洪财
孙秋
XING Wei;CHENG Yanyan;WANG Shuren;WANG Yanli;WANG Wenran;ZHANG Hongcai;SUN Qiu(The First Affliated Hospital,Heilongjiang University of Chinese Medicine,Harbin 150040,China;Heilongjiang Nursing College,Harbin 150301,China;Research Institute of Traditional Chinese Medicine,Heilongjiang University of Chinese Medicine,Harbin 150040,China)
出处
《广东药科大学学报》
CAS
2020年第2期254-258,共5页
Journal of Guangdong Pharmaceutical University
基金
黑龙江省中医药管理局科研项目(2018-53)。
