TAN Sihang, SHI Jixiang, ZHOU Qiang, ZHOU Junjie, HAO Shengkun,WANG Wenyan, ZHUANG Weikang
（Department of Orthopadics, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200062, China）
Abstract： When gene expression profile is used for gene detection, the probe on the chip can emit fluorescence with different wavelengths. Under the action of confocal laser scanner, a clear gene change image can be obtained, by which the gene changes of the sample to be tested can be observed directly. First, the knee osteoarthritis (KOA) models of mice are established by the method of collateral ligament and meniscus resection (MLI-OA). Then, Bushen Huoxue formula is given by gavage, and ribonucleic acid (RNA) is routinely extracted and purified. Finally, the gene expression changes of KOA tissues of mice are detected by Agilent SurePrint G3 Mouse GE V2.0 gene expression profile. The results show that Bushen Huoxue formula has significant regulation effect on gene expression of KOA tissue. Among the genes with significant up-regulation effect of Bushen Huoxue formula, there are 56 genes of traditional Chinese medicine (TCM) groups up-regulated more than twice compared with model groups. Among the genes with significant down-regulation effect, there are 119 genes of TCM groups down-regulated more than twice compared with model groups. The experimental results indicate that Bushen Huoxue formula may promote the metabolism of arthritic factors and delay cartilage degeneration to treat KOA by regulating genes that are currently unknown in the pathological process of KOA.
Key words： gene expression profile； Bushen Huoxue formula； knee osteoarthritis (KOA)； gene chip； mice
References
［1］Conaghan P G, Peloso P M, Everett S V, et al. Inadequate pain relief and large functional loss among patients with knee osteoarthritis： evidence from a prospective multinational longitudinal study of osteoarthritis real-world therapies. Rheumatology， 2015， 54： 270-277.
［2］Blackburn A R, Hamrick M W, Chutkan N, et al. Comparative analysis of sodium coupled vitamin c transporter 2 in human osteoarthritis grade 1 and grade 3 tissues. BMC Musculoskeletal Disorders， 2014， 15： 9
［3］Aref-Eshghi E, Zhang Y, Liu M， et al. Genome-wide DNA methylation study of hip and knee cartilage reveals embryonic organ and skeletal system morphogenesis as major pathways involved in osteoarthritis. BMC Musculoskeletal Disorders， 2015， 16： 287.
［4］Allen K D, Bosworth H B, Brock D S, et al. Patient and provider interventions for managing osteoarthritis in primary care： Protocols for two randomized controlled trials. BMC Musculoskeletal Disorders， 2012， 13： 60.
［5］Jordan K P, Jud A, Bergknut C, et al. International comparisons of the consultation prevalence of musculoskeletal conditions using population-based healthcare data from England and Sweden. Annals of the Rheumatic Diseases， 2014， 73(1)： 212-218 .
［6］Silverwood V, Blagojevic-Bucknall M, Jinks C, et al. Current evidence on risk factors for knee osteoarthritis in older adults： a systematic review and meta-analysis. Osteoarthritis Cartilage， 2015， 23(4)： 507-521.
［7］Fibel K H, Hillstrom H J, Halpern B C. State-of-the-art management of knee osteoarthritis.World Journal of Clinical Cases， 2015， 3(2)： 89-101
［8］Wang SP, Wang D L, Li J R, et al. Identification and analysis of the cleavage site in a signal peptide using SMOTE, dagging, and feature selection methods. Molecular Omics， 2018， 14(1)： 64-73.
［9］Khan S, Nasee I, Togneri K, et al. RAFP-Pred： robust prediction of antifreeze proteins using localized analysis of n-peptide compositions, IEEE/ACM Transactions on Computational Biology & Bioinformatics， 2018, 15 (1)： 244-250.
［10］Braunstein M J, Kucharczyk J, Adams S. Targeting toll-like receptors for cancer therapy. Targeted Oncology， 2018, 13： 583-598.
［11］Vainieri M L, Wahl D, Alini M, et al. Mechanically stimulated osteochondral organ culture for evaluation of biomaterials in cartilage repair studies, Acta Biomater，2018, 81： 256-266.
［12］Liu W, Zhang G, Wu J, et al. Insights into the angiogenic effects of nanomaterials： mechanisms involved and potential applications, Journal of Nanobiotechnology， 2020, 18： 9.
［13］Shi J, Liang Q, Zuscik M, et al. Distribution and alteration of lymphatic vessels in knee joints of normal and osteoarthritic mice. Arthritis and Rheumatology， 2014, 66(3)： 657-666.
［14］Sampson E R, Beck C A, Ketz J, et al. Establishment of an index with increased sensitivity for assessing murine arthritis. Journal of Orthopaedic Research, 2011, 29： 1145-1151.
［15］Liu W， Wu Y H， Liu X Y， et al．Metabolic regulatory and anti-oxidative effects of modified Bushen Huoxue Decoction on experimental rabbit model of osteoarthritis. Chinese Journal of Integrative Medicine， 2013， 19(6)： 459-463.
［16］Yan C, Li J, Qiang L, et al. Du-Huo-Ji-Sheng-Tang attenuates inflammation of TNF-Tg mice related to promoting lymphatic drainage function. Evidence-based Complementray and Alternative Medicine, 2016(3)： 7067691.
基因表达谱芯片用于小鼠膝关节炎组织的检测与分析
谭思航， 石继祥， 周强， 周军杰， 郝胜坤， 王文燕， 庄伟康
（上海中医药大学 附属普陀医院， 上海 200062）
摘要： 通过副韧带及半月板切除方法（MLI-OA）建立小鼠KOA模型， 补肾活血方灌胃给药、 常规抽提和纯化RNA， 采用Agilent SurePrint G3 Mouse GE V2.0基因表达谱芯片检测小鼠膝OA关节组织的基因表达变化。 检测结果显示补肾活血方对KOA组织基因表达具有明显调控作用：补肾活血方有明显上调作用的基因中， 中药组比模型组上调2倍以上的有56个； 明显下调作用的基因中， 中药组比模型组下调２倍以上的有119个。 研究结果表明补肾活血方可能通过调控目前在KOA病变过程中作用尚不明确的基因， 促进关节炎性因子代谢， 延缓软骨退变， 治疗骨关节炎的作用。
关键词： 基因表达谱检测； 补肾活血方； 膝关节炎； 基因芯片； 小鼠
引用格式： TAN Sihang, SHI Jixiang, ZHOU Qiang, et al. Detection and analysis of knee osteoarthritis in mice with gene expression profile. Journal of Measurement Science and Instrumentation, 2021, 12（1）： 115-119. DOI： 10．3969／j．issn．1674-8042．2021．01．015
[full text view]
