鹿仲健骨汤治疗股骨头坏死模型大鼠:肠道菌群及血清激素变化  

Luzhongjiangu decoction for the treatment of femoral head necrosis in rats:changes in intestinal flora and serum hormones

作　　者：景田园 王平[2] 王仪 胡亚楠 刘善新[2] 孙国栋 杜海涛 Jing Tianyuan;Wang Ping;Wang Yi;Hu Yanan;Liu Shanxin;Sun Guodong;Du Haitao(Shandong Xiandai University,Jinan 250104,Shandong Province,China;Shandong Academy of Chinese Medicine,Jinan 250014,Shandong Province,China;Jinan Biopharmaceutical Industry Research Institute,Jinan 250000,Shandong Province,China;The Third Affiliated Hospital of Shandong first Medical University,Jinan 250031,Shandong Province,China)

机构地区：[1]山东现代学院药学院,山东省济南市250104 [2]山东省中医药研究院,山东省济南市250014 [3]济南生物医药产业研究院,山东省济南市250000 [4]山东第一医科大学第三附属医院,山东省济南市250031

出　　处：《中国组织工程研究》2024年第35期5598-5605,共8页Chinese Journal of Tissue Engineering Research

基　　金：山东省重点研发计划(科技示范工程)(2021SFGC1205),项目负责人:王平;山东省自然科学基金(ZR2020MH386),项目负责人:王平;中央引导地方科技发展专项基金(YDZX2021117),项目负责人:王平;济南市“新高校20条”资助项目(202228121),项目负责人:王平;山东省中央引导地方科技发展资金项目(YDZX20203700002055),项目负责人:王平。

摘　　要：背景:股骨头坏死是一种常见的致残性疾病,主要表现为微循环障碍、骨细胞代谢紊乱。鹿仲健骨汤由山东省中医药研究院研制而成,在临床以汤剂形式使用,对股骨头坏死具有良好疗效,然而其作用机制尚未明确。目的:基于16S rDNA技术研究鹿仲健骨汤对股骨头坏死模型大鼠肠道菌群的影响,探究其治疗股骨头坏死的作用机制。方法:采用维甲酸灌胃的方式建立Wistar大鼠股骨头坏死模型,通过股骨头部位骨形态计量学、骨组织病理学和血清激素水平评价鹿仲健骨汤的治疗作用,采用16S rDNA测序技术对空白对照组、模型组及鹿仲健骨汤中剂量组大鼠肠道菌群进行分析,构建相应文库并进行OTU聚类以及微生物群落多样性、丰度等分析,明确肠道菌群的组成及各组间的菌群种类、多样性变化。结果与结论:(1)鹿仲健骨汤可以显著提高骨钙素、骨桥蛋白等成骨相关因子的表达,缓解骨小梁的破坏,增加骨密度,对股骨头坏死具有显著的治疗作用,鹿仲健骨汤中剂量组效果最为显著;(2)肠道菌群测序结果显示,鹿仲健骨汤在一定程度上改善了股骨头坏死大鼠菌群紊乱,共筛选出杆菌门、脱硫弧菌门、脱硫孤菌属、异杆菌属、双歧杆菌属等差异菌落;能上调双歧杆菌等有益菌丰度,下调脱硫弧菌等有害菌丰度,改善肠道菌群结构;(3)通过功能预测分析发现鹿仲健骨汤治疗股骨头坏死可能主要影响氨基酸代谢、能量代谢等;(4)相关性分析显示,鹿仲健骨汤中剂量组的差异菌双歧杆菌属(Bifidobacterium)、肠单胞球菌(Intestinimonas)与维生素D3、雌二醇、降钙素呈正相关,与前列腺素E2呈负相关;模型组的差异菌大肠杆菌-志贺氏菌(Escherichia-Shigella)、脱硫孤菌属(Desulfovibrio)、格鲁比卡氏菌属(Globicatella)、链球菌属(Streptococcus)与前列腺素E2呈正相关,与维生素D3、雌二醇、降钙素呈负相关。结果表�BACKGROUND:Osteonecrosis of the femoral head is a common and disabling disease,which is mainly characterized by microcirculation disorders and bone cell metabolism disorders.Luzhongjiangu decoction was developed by Shandong Academy of Chinese Medicine and used in the form of soup in the clinic,which has good efficacy in the treatment of osteonecrosis of the femoral head.However,its mechanism of action has not been clarified.OBJECTIVE:To study the effect mechanism of Luzhongjiangu decoction on intestinal flora in rats with osteonecrosis of the femoral head based on 16S rDNA sequencing technique.METHODS:The model of osteonecrosis of the femoral head was established in Wistar rats by intragastric administration of retinoic acid.The therapeutic effect of Luzhongjiangu decoction was evaluated by serum hormone,bone histopathology and serum hormone levels.16s rDNA sequencing technique was used to detect the intestinal flora of rats in the blank control group,model group and middle-dose Luzhongjiangu decoction group.The corresponding library was constructed and OTU clustering and microbial community diversity and abundance analysis were carried out to determine the composition of intestinal flora and the changes of species and diversity among groups.RESULTS AND CONCLUSION:Luzhongjiangu decoction could significantly increase the expression of osteocalcin,osteopontin and other osteogenic related factors,alleviate the destruction of bone trabeculae,increase bone mineral density,and had a significant therapeutic effect on osteonecrosis of the femoral head,of which the middle dose group showed the most significant effect.The results of intestinal flora sequencing showed that Luzhongjiangu decoction improved the flora disorder of rats with osteonecrosis of the femoral head to some extent,and screened out different colonies such as Bacillus,Desulfurizans,Desulfurization,Isobacteria,Bifidobacterium and so on;it could up-regulate the abundance of beneficial bacteria such as Bifidobacterium,down-regulate the abundance of harmful b

关 键 词：鹿仲健骨汤 股骨头坏死 16S rDNA 肠道菌群 激素 维生素D3 雌二醇 降钙素 

分 类 号：R496[医药卫生—康复医学] R318[医药卫生—临床医学] R446