补肾化痰方对去卵巢大鼠骨形成的影响及其作用机制研究  

Effect of Bushen Huatan Fang( 补肾化痰方) on bone formation of ovariectomized rats and its mechanism of action

作　　者：张麟 向楠[2] 周广文[2] 周亚娜[3] 胡娅[4] 张妍 李章青 ZHANG Lin;XIANG Nan;ZHOU Guangwen;ZHOU Yana;HU Ya;ZHANG Yan;LI Zhangqing(The First Hospital of Wuhan,Wuhan 430022,Hubei,China;Hubei University of Traditional Chinese Medicine,Wuhan 430061,Hubei,China;Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan 430061,Hubei,China;Changjiang University,Jingzhou 434023,Hubei,China)

机构地区：[1]湖北省武汉市第一医院,湖北武汉430022 [2]湖北中医药大学,湖北武汉430061 [3]湖北省中医院,湖北武汉430061 [4]长江大学,湖北荆州434023

出　　处：《中医正骨》2018年第10期12-18,共7页The Journal of Traditional Chinese Orthopedics and Traumatology

基　　金：湖北省卫生厅中医药中西医结合科研项目(2012Z-Y31);湖北中医药大学"青苗计划"资助项目(2016ZZX016)

摘　　要：目的:观察补肾化痰方对去卵巢大鼠骨形成的影响及其作用机制。方法:将30只6月龄SPF级雌性SD大鼠随机分为假手术组、模型组和补肾化痰组,每组10只。模型组和补肾化痰组大鼠手术摘除双侧卵巢,假手术组不摘除卵巢,只切除卵巢周围与双侧卵巢质量相等的脂肪组织。造模术后5周开始,补肾化痰组以0. 94 g·m L^(-1)补肾化痰方药液灌胃,假手术组和模型组以等体积生理盐水灌胃,均每天1次,连续干预8周。药物干预结束后,将所有大鼠处死,摘除双侧眼球取血检测血清骨形态发生蛋白(bone morphogenetic protein,BMP)-2、Runt相关转录因子(Runt related transcription factor,RUNX)-2含量,取左侧股骨行Micro CT扫描测定骨密度、骨小梁数量、骨小梁平均厚度、骨小梁分离度,取右侧股骨上端部分骨片以蛋白免疫印迹法测定骨组织中BMP-2、RUNX-2含量,取右侧股骨下端部分骨片以免疫组织化学法检测BMP-2、RUNX-2的定位。结果:(1)一般情况。所有大鼠均在造模手术麻醉3~4 h后完全清醒,正常进食,1周后切口完全愈合。药物干预过程中,模型组1只大鼠死亡、补肾化痰组2只大鼠死亡。(2)血清BMP-2、RUNX-2含量检测结果。3组大鼠的血清BMP-2含量比较,差异有统计学意义[(245. 40±10. 10) pg·m L^(-1),(111. 03±29. 75) pg·mL^(-1),(190. 35±14. 00) pg·mL^(-1),F=109. 998,P=0. 000];模型组、补肾化痰组的血清BMP-2含量均低于假手术组(P=0. 000,P=0. 000),补肾化痰组的血清BMP-2含量高于模型组(P=0. 000)。3组大鼠的血清RUNX-2含量比较,差异有统计学意义[(4 131. 04±277. 39) pg·mL^(-1),(1 601. 09±266. 10) pg·mL^(-1),(3 134. 90±202. 33) pg·mL^(-1),F=237. 192,P=0. 000];模型组、补肾化痰组的血清RUNX-2含量均低于假手术组(P=0. 000,P=0. 000),补肾化痰组的血清RUNX-2含量高于模型组(P=0. 000)。(3)股骨Micro CT扫描结果。3组大鼠骨密度比较,差异有统计学意义[(556. 90±8. 86) mg HA·cmObjective:To observe the effects of Bushen Huatan Fang(补肾化痰方,BSHTF)on bone formation of ovariectomized rats and its mechanism of action.Methods:Thirty 6-month-old SPF-grade female SD rats were randomly divided into sham-operated group,model group and BSHTF group,10 cases in each group.The bilateral ovariectomy were performed on rats in model group and BSHTF group,and the resection of same mass of fat around bilateral ovaries were performed on rats in sham-operated group.Since the 5th week after the modeling operation,the rats in BSHTF group were intragastric administrated with BSHTF solution in dosages of 0.94 g/mL,while the rats in sham-operated group and model group were intragastric administrated with the same dose of normal saline,once a day for consecutive 8 weeks.After the end of drug intervention,all rats were executed and their bilateral eyeballs were removed and their blood were fetched out for detecting the serum contents of bone morphogenetic protein(BMP)-2 and Runt related transcription factor(RUNX)-2.Their left femurs were fetched out for determining bone mineral density(BMD),trabecular number(Tb.N),mean trabecular thickness(Tb.Th)and trabecular separation(Tb.Sp)through Micro-CT scanning.Meanwhile,the sclerites were fetched out from the upper end and the lower end of right femurs respectively for measuring the contents of BMP-2 and RUNX-2 in bone tissues by using Western blot assay and detecting the location of BMP-2 and RUNX-2 by using immunohistochemical method.Results:All rats were fully waked up from 3-4-hour anesthesia for the modeling surgery and fed normally,and their incisions healed completely 1 week later.One rat in model group and 2 rats in BSHTF group died during the drug intervention.The detection results of serum contents of BMP-2 and RUNX-2 showed that there was statistical difference in serum contents of BMP-2 between the 3 groups(245.40+/-10.10,111.03+/-29.75,190.35+/-14.00 pg/mL,F=109.998,P=0.000).The serum contents of BMP-2 were lower in model group and BSHTF group compar

关 键 词：骨质疏松 补肾化痰方 信号传导 骨形态发生蛋白质2 Runt相关转录因子2 大鼠 Sprague-Dawley 动物实验 

分 类 号：R285.5[医药卫生—中药学]