加减知柏地黄丸对特发性中枢性性早熟小鼠Runx2、OSX表达的影响  

Effects of Modified Zhibai Dihuang Pill on the expression of Runx2 and OSX in idiopathic central precocious puberty mice

作　　者：黄桢 王修银 叶玉莲 曾博煌 张泳 HUANG zhen;WANG Xiuyin;YE Yulian(Laboratory of Affiliated Hospital of Traditional Chinese Medicine,Guangzhou Medical University,Guangdong Guangzhou 510130)

机构地区：[1]广州医科大学附属中医医院检验科,广东广州510130

出　　处：《医学检验与临床》2023年第1期5-9,44,共6页Medical Laboratory Science and Clinics

基　　金：广东省中医药局科研项目,项目编号:20201258;广州市中医药和中西医结合科技项目,项目编号:20212A011006。

摘　　要：目的:观察加减知柏地黄丸对特发性中枢性性早熟(ICPP)小鼠核心结合蛋白因子2(Runx2)、成骨细胞特异性转录因子(OSX)的影响,探讨加减知柏地黄丸治疗ICPP缓解骨骺提前融合的作用机制。方法:本实验采用腹腔注射瘦素(Leptin)(2 mg/kg)SPF级雌性C57小鼠复制特发性中枢性性早熟(ICPP)模型,将小鼠随机分成6组:正常对照组、ICPP模型对照组、加减知柏地黄丸高剂量组、加减知柏地黄丸低剂量组、甲地孕酮对照组、GnRH-A组。正常雌性小鼠阴道开口时间一般在日龄26~28 d,以27 d较为集中,因此设计为小鼠26 d日龄时结束实验;小鼠实验结束前1 d,ELISA检测骨生化指标碱性磷酸酶(ALP)、胰岛素样生长因子1(IGF-1)、N-MID骨钙素片段(N-MID)和激素黄体生成激素(LH)、卵泡刺激素(FSH),Real-Time PCR、Western blot检测Runx2、OSX在ICPP骨组织中的表达水平。结果:与正常组比较,ICPP模型组骨生化指标ALP、IGF-1、N-MID以及激素LH和FSH浓度明显升高(P<0.01),Runx2、OSX表达水平明显增高(P<0.01)。与模型组比较,加减知柏地黄丸和甲地孕酮对照组可以明显下调ALP、IGF-1、N-MID和激素LH和FSH浓度,以及RUNX2、OSX表达水平(P<0.05,P<0.01),并与加减知柏地黄丸剂量相关。结论:加减知柏地黄丸是通过作用于Runx2、OSX骨信号通路,改变血清中骨相关指标和激素的水平,来减慢骨龄的进展,缓解骨骺提前融合。Objective:To observe the influence of Modified Zhibai Dihuang Pill on core binding protein factor 2(Runx2)and osteoblast specific transcription factor(OSX)in mice with idiopathic central precocious puberty(ICPP)and to explore the mechanism of Modified Zhibai Dihuang Pill in the treatment of ICPP in relieving the premature fusion of the epiphysis.Methods:ICPP model was replicated in SPF female C57 mice by intraperitoneal injection of leptin(2 mg/kg).The mice were randomly divided into 6 groups:normal control group,ICPP model control group,Modified Zhibai Dihuang Pill high-dose group,Modified Zhibai Dihuang Pill low-dose group,megestrol control group and GnRH-A group.The vaginal opening time of normal female mice was generally 26~28 days old.Therefore,it was designed to end the experiment when the mice were 26 days old.One day before the end,alkaline phosphatase(ALP),insulin like growth factor-1(IGF-1),N-MID osteocalcin fragment(N-MID),luteinizing hormone(LH),follicle stimulating hormone(FSH)were measured by ELISA.The expression levels of Runx2 and OSX in ICPP bone tissue were measured by real time PCR and Western blot.Results:Comparing with the normal group,the concentrations of ALP,IGF-1,N-MID,LH and FSH in ICPP model group were significantly higher(P<0.01),and the expression levels of Runx2 and OSX were significantly higher(P<0.01).Comparing with the model group,the levels of ALP,IGF-1,N-MID,LH and FSH increased significantly in Modified Zhibai Dihuang pill groups and megestrol control group,as well as the expression levels of Runx2 and OSX(P<0.05,P<0.01).In addition,the drop is related to the dosage of Modified Zhibai Dihuang pill.Conclusion:Modified Zhibai Dihuang Pill acts on the Runx2 and OSX bone signaling pathways and changes the levels of bone-related indexes and hormones in serum to slow down the progression of bone age and relieve the premature fusion of the epiphysis.

关 键 词：加减知柏地黄丸 RUNX2 OSX 

分 类 号：R285.5[医药卫生—中药学]