基于转录组测序技术探究电针促进新西兰兔骨缺损愈合的作用机制  

Mechanism of electroacupuncture in promoting bone defect healing in New Zealand rabbits based on transcriptome sequencing technology

作　　者：李霞 周立群[2] 徐林[1] 王兴建 田向东 王立 耿少辉 蒋海旭 许恒 赵吉平[1] LI Xia;ZHOU Liqun;XU Lin;WANG Xingjian;TIAN Xiangdong;WANG Li;GENG Shaohui;JIANG Haixu;XU Heng;ZHAO Jiping(Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China;Wudang Medical Research Institute,Beijing University of Chinese Medicine,Beijing 100029,China;The Third Affiliated Hospital of Beijing University of Chinese Medicine,Beijing 100029,China;Anyuan Quanlu(Beijing)Institute of Medical Sciences,Beijing 100055,China;School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102446,China)

机构地区：[1]北京中医药大学东直门医院,北京100700 [2]北京中医药大学武当医学研究院,北京100029 [3]北京中医药大学第三附属医院,北京100029 [4]安元全律(北京)医学研究院,北京100055 [5]北京中医药大学生命科学学院,北京102446

出　　处：《现代中西医结合杂志》2022年第16期2211-2217,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国防科技创新特区H863计划项目(20-163-02-ZT-008-003-01)。

摘　　要：目的基于转录组测序技术观察电针促进骨折愈合过程中肾组织基因表达谱变化,揭示电针促进骨折愈合的分子生物学机制。方法将42只新西兰兔随机分为正常组6只、骨缺损组18只、电针组18只。骨缺损组与电针组使用咬骨钳在新西兰兔右前肢桡骨中段造成骨缺损模型。造模后,电针组在骨缺损肢体侧的合谷、曲池穴处,采用疏密波(频率2/100 Hz,电流2.1 mA)进行电针干预,30 min/次,1次/d,连续干预4周。分别在实验1,2,4周末对骨缺损组与电针组兔前肢进行X射线平扫,采用Lane-Sandhu X射线评分评估骨折愈合情况;在实验2周末每组随机选取3只兔,进行肾脏组织转录组测序以检测基因表达情况。结果实验1,2,4周末,电针组X射线骨形成评分、骨连接评分、骨塑形评分均较同期骨缺损组高,但仅2周末时X射线骨塑形评分与骨缺损组比较差异有统计学意义(P<0.05)。转录组测序显示:正常组与骨缺损组间有454个差异表达基因(上调基因255个,下调基因199个),骨缺损组与电针组间有444个差异表达基因(上调基因179个,下调基因265个)。钙感应受体(CASR)基因在骨缺损组和电针组的肾脏中差异表达,GO分析发现骨缺损组与电针组的差异基因富集在钙离子稳态和细胞钙离子稳态等生物学过程中。结论电针可显著促进新西兰兔骨折愈合,其作用机制可能与调节肾脏CASR基因表达和“钙离子稳态”通路有关。Objective It is to observe the changes of gene expression profile of renal tissue during promoting fracture healing by electroacupuncture based on transcriptome sequencing technology,and to reveal the molecular biological mechanism of electroacupuncture promotion for fracture healing.Methods Forty-two New Zealand rabbits were randomly divided into normal group(n=6),bone defect group(n=18)and electroacupuncture group(n=18).Bone defect models were established in the mid-radius of the right forelimbs of the New Zealand rabbits using a rongeur in the bone defect group and the electroacupuncture group.After modeling,the electroacupuncture group was treated with electroacupuncture intervention at Hegu and Quchi acupoints on the side of the limb with bone defect,using sparse and dense waves(frequency 2/100 Hz,current 2.1 mA),30 min/time,1 time/d,continuously treated for 4 weeks.The forelimbs of the rabbits in the bone defect group and the electroacupuncture group were respectively scanned by X-ray at the end of the 1st,2nd,and 4th weekends of the experiment,and the fracture healing was evaluated by Lane-Sandhu X-ray score.At the end of the second week of the experiment,3 rabbits were randomly selected from each group to perform transcriptome sequencing of kidney tissue to detect gene expression.Results At the end of the 1st,2nd,and 4th week of the experiment,the X-ray bone formation score,bone connection score and bone remodeling score of the electroacupuncture group were all higher than those of the bone defect group at the same period,but the X-ray bone remodeling score was significantly different from the bone defect group only at the 2nd weekend(P<0.05).Transcriptome sequencing showed that there were 454 differentially expressed genes(255 up-regulated genes,199 down-regulated genes)between the normal group and the bone defect group,and 444 differentially expressed genes(179 up-regulated genes,265 down-regulated genes)between the bone defect group and the electroacupuncture group.Calcium sensing receptor(CASR)genes w

关 键 词：骨缺损 电针 转录组测序 钙感应受体 钙离子稳态 

分 类 号：R-332[医药卫生]