加减知柏地黄丸对特发性中枢性性早熟小鼠骨细胞骨形成蛋白-Smads信号通路的影响  

Study on the effect of modified Zhibai Dihuang pill on bone cells in idiopathic central precocious puberty mice based on BMP-Smads signaling pathway

作　　者：何敏 黄桢 He Min;Huang Zhen(Department of Anesthesiology,Southern Theater General Hospital of the Chinese People's Liberation Army,Guangzhou Guangdong 510010,China;Laboratory of Affiliated Hospital of Traditional Chinese Medicine,Guangzhou Medical University,Guangzhou Guangdong 510130,China)

机构地区：[1]中国人民解放军南部战区总医院麻醉科,广东广州510010 [2]广州医科大学附属中医医院检验科,广东广州510130

出　　处：《中华临床实验室管理电子杂志》2023年第4期214-220,共7页Chinese Journal of Clinical Laboratory Management(Electronic Edition）

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

摘　　要：目的探讨加减知柏地黄丸对特发性中枢性性早熟(ICPP)小鼠骨细胞骨形成蛋白(BMP)-Smads信号通路的影响。方法将36只SPF级雌性C57小鼠随机分为6组:正常对照组、ICPP模型组、知柏地黄丸高剂量组、知柏地黄丸低剂量组、甲地孕酮阳性对照组、促性腺激素释放激素类似物(GnRH-A)组,每组6只;正常对照组给予生理盐水腹腔注射,同时给予蒸馏水灌胃,上午给药,每天1次,连续7 d。ICPP模型组和用药组把腹腔注射更换为瘦素,注射药量为2 mg/kg,ICPP模型组用蒸馏水灌胃,知柏地黄丸高剂量组、知柏地黄丸低剂量组、甲地孕酮阳性对照组用对应的药物灌胃,连续7 d,每天1次;GnRH-A组不灌胃而是以抑那通于右后肢处肌肉注射,隔日1次,共3次。所有组别于小鼠日龄26 d时结束实验。取下腔静脉血,采用酶联免疫吸附法(ELISA)检测血清中促卵泡素(FSH)、促黄体激素(LH)、N端中段骨钙素(N-MID)、碱性磷酸酶(ALP)、胰岛素样生长因子1(IGF-1);取右侧股骨免疫印迹检测BMP-2、Smad1、Smad5、Smad8与Smad4表达量。结果与正常对照组比较,ICPP模型组血清中FSH、LH、N-MID、ALP、IGF-1含量明显升高(P<0.05),BMP-2、Smad1、Smad5、Smad8与Smad4表达量明显上调(P<0.05)。与ICPP模型组比较,知柏地黄丸高剂量组、知柏地黄丸低剂量组、甲地孕酮阳性对照组中FSH、LH、N-MID、ALP、IGF-1含量明显降低(P<0.05),BMP-2、Smad1、Smad5、Smad8与Smad4表达量明显降低(P<0.05),知柏地黄丸高剂量组比低剂量组下调明显。与ICPP模型组比较,GnRH-A组FSH、LH含量偏高(P<0.05),ALP含量偏低(P<0.05),N-MID、IGF-1差异无统计学意义,BMP-2、Smad1、Smad5与Smad4表达量差异有统计学意义(P<0.05),Smad8差异无统计学意义。结论ICPP小鼠骨细胞BMP-Smads信号通路表达增强,加减知柏地黄丸可以下调BMP-Smads信号通路的表达水平,且与知柏地黄丸具有剂量相关性。Objective To investigate the effect of modified Zhibai Dihuang pill on the bone morphogenetic protein(BMP)-Smads signaling pathway in bone cells from idiopathic central precocious puberty(ICPP)mice.Methods 36 SPF-grade female C57 mice were randomly divided into six groups:normal control group,ICPP model group,high-dose Zhibai Dihuang pill group,low-dose Zhibai Dihuang pill group,megestrol acetate positive control group,and gonadotropin releasing hormone analogue(GnRH-A)group.The normal control group was administered with physiological saline by intraperitoneal injection and distilled water was administered by gavage in the morning,once a day for 7 consecutive days.The ICPP model group and the drug group were administered with leptin by intraperitoneal injection at a dose of 2 mg/kg.The Zhibai Dihuang pill groups,the medroxyprogesterone acetate positive control group,and the GnRH-A group were administered with the corresponding drugs by gavage,also once a day for 7 days.The GnRH-A group received intramuscular injection of Yinatong at the right hind limb without gavage,once every other day,for a total of three times.The experiment was ended at 26 days.Serum follicle-stimulating hormone(FSH),luteinizing hormone(LH),N-mid osteocalcin(N-MID),alkaline phosphatase(ALP),and insulin-like growth factor-1(IGF-1)were measured using enzyme linked immunosorbent assay(ELISA)in inferior vena cava blood.Western blot was detected the expression of BMP-2,Smad1,Smad5,Smad8,and Smad4 in the right femur.Results Compared with the normal control group,the serum levels of FSH,LH,N-MID,ALP,and IGF-1 in the ICPP model group were significantly increased(P<0.05),and the expression levels of BMP-2,Smad1,Smad5,Smad8,and Smad4 were significantly increased(P<0.05);Compared with the ICPP model group,the levels of FSH,LH,N-MID,ALP,and IGF-1 in the high-dose group of Zhibai Dihuang pill,the low-dose group of Zhibai Dihuang pill,and the megestrol positive control group were significantly decreased(P<0.05),the expression of BMP-2,Smad1,Smad5,Smad8 an

关 键 词：加减知柏地黄丸 特发性中枢性性早熟 骨形成蛋白 SMADS 信号通路 

分 类 号：R285.5[医药卫生—中药学]