少阳主骨方对食蟹猴自然退变膝关节骨性关节炎模型p16/Rb信号通路的影响

doi:10.13418/j.issn.1001-165x.2017.06.013

中国临床解剖学杂志 ›› 2017, Vol. 35 ›› Issue (6): 655-660.doi: 10.13418/j.issn.1001-165x.2017.06.013

少阳主骨方对食蟹猴自然退变膝关节骨性关节炎模型p16/Rb信号通路的影响

周鑫，　张磊，　扶世杰，　刘刚，　郭晓光，　汪国友

西南医科大学附属中医医院骨科，四川泸州 646000

收稿日期:2017-07-13 出版日期:2017-11-25 发布日期:2017-12-30
通讯作者: 汪国友，医学博士，副教授，Tel:18715799097，E-mail:285522951@qq.com
作者简介:周鑫（1992-），男，四川会理人，硕士研究生，主要从事骨关节基础与临床研究，Tel:18715799097，E-mail:285522951@qq.com
基金资助:
四川省科技支撑计划项目课题项目(2014SZ0185)；泸州市科技计划项目课题项目（2013LZLY-K59）；2016年西南医科大学-西南医科大学附属中医医院联合专项项目（2016-4-4）；2016年泸州市-西南医科大学科技战略合作项目（2016LZXNYD-J08）

Effects of ShaoYangZhuGuFang on p16/Rb signaling pathway of naturally degenerative cynomolgus monkeys knee osteoarthritis model

ZHOU Xin, ZHANG Lei, FU Shi-jie, LIU Gang, GUO Xiao-guang, WANG Guo-you

Department of orthopedics, Affiliated T.C.M Hospital of Southwest Medical University, Luzhou 646000, China

Received:2017-07-13 Online:2017-11-25 Published:2017-12-30

摘要/Abstract

摘要：

目的　根据中医“少阳主骨”理论，研究少阳主骨方延缓关节软骨细胞衰老和退变的作用机制。方法　通过X线选取13只膝关节退变的老年食蟹猴作为动物模型，随机选取1只进行病理学观察，其余食蟹猴随机分为少阳主骨方组、氨糖美辛组、生理盐水组，每组4只，连续灌胃8周后处死所有食蟹猴，对关节软骨进行病理学观察，RT-qPCR、Western-blot检测关节软骨中p16、Rb基因与蛋白的表达。结果　老年食蟹猴KOA模型关节软骨病理改变符合KOA病理变化特点，三组食蟹猴Mankin评分少阳主骨方组(7.5±0.53)分、氨糖美辛组(7.75±0.71)分、生理盐水组(8.25±0.46)分，少阳主骨方组与生理盐水组间具有统计学差异（P<0.05）；p16基因与蛋白的表达少阳主骨方组<氨糖美辛组<生理盐水组，具有统计学差异（P<0.05）；Rb基因与蛋白的表达少阳主骨方组>氨糖美辛组>生理盐水组，具有统计学差异（P<0.05）。结论　少阳主骨方可以通过p16/Rb途径延缓关节软骨细胞衰老，从而延缓关节软骨退变。

关键词: 少阳主骨, 　食蟹猴膝关节骨性关节炎模型, 　细胞衰老, 　p16/Rb信号通路

Abstract:

Objective　 The aim of this study was to investigate the mechanism of ShaoYangZhuGuFang(SYZGF) in delaying the cell senescence and degeneration of articular chondrocytes according to the theory of "ShaoYangZhuGu". Method 13 cynomolgus monkeys with degenerative knee joints were selected by X-ray. One of the monkeys was selected randomly for pathologic observation, and the other cynomolgus monkeys were randomly divided into SYZGF group, Ammonia Moxime group and Saline group. There were four cynomolgus monkeys in each group. All cynomolgus monkeys were executed after 8 consecutive weeks of intragastric administration. Pathological observation of articular cartilage was made and the expression of p16, Rb gene and protein in articular cartilage was detected by RT-qPCR and Western-blot technique.　Result　Articular cartilage pathological changes of elderly cynomolgus monkey KOA models was consistent with the characteristics of KOA pathological changes. Mankin scores of cynomolgus monkeys in three groups were as follows: 7.5 ± 0.53 in SYZGF group, 7.75 ± 0.71 in Ammonia Moxime group, 8.25 ± 0.46 in Saline group, and there was significant difference between SYZGF group and Saline group (P<0.05). The expression of p16 was lowest in SYZGF group, which in middle in Ammonia Moxime group, and highest in Saline group. There was significant difference in the three groups (P<0.05). The expression of Rb was highest in SYZGF group, middle in Ammonia Moxime group, and lowest in Saline group. There was significant difference in the three groups (P<0.05).　Conclusion　SYZGF could delay chondrocyte senescence by p16/Rb pathway and delay articular cartilage degeneration.

Key words: ShaoYangZhuGu, 　Cynomolgus monkeys knee osteoarthritis model, 　Cell senescence, p16/Rb signaling pathway

周鑫,张磊,扶世杰,刘刚,郭晓光,汪国友. 少阳主骨方对食蟹猴自然退变膝关节骨性关节炎模型p16/Rb信号通路的影响[J]. 中国临床解剖学杂志, 2017, 35(6): 655-660.

ZHOU Xin, ZHANG Lei, FU Shi-jie, LIU Gang, GUO Xiao-guang, WANG Guo-you. Effects of ShaoYangZhuGuFang on p16/Rb signaling pathway of naturally degenerative cynomolgus monkeys knee osteoarthritis model[J]. Chinese Journal Of Clinical Anatomy, 2017, 35(6): 655-660.

参考文献

[1] Alayat MS, Aly TH, Elsayed AE. Efficacy of pulsed Nd: YAG laser in the treatment of patients with knee osteoarthritis: a randomized controlled trial[J]. Lasers Med Sci, 2017, 32(3): 503-511.
[2] Riis RG, Gudbergsen H, Simonsen O, et al. The association between histological, macroscopic and magnetic resonance imaging assessed synovitis in end-stage knee osteoarthritis: a cross-sectional study[J]. Osteoarthritis Cartilage, 2017, 25(2): 272-280.
[3] Michael JW, Schlüter-Brust KU, Eysel P. The epidemiology, etiology, diagnosis, and treatment of osteoarthritis of the knee[J]. Dtsch Arztebl Int, 2010, 107(9): 152-62.
[4] Gouttebarge V, Inklaar H, Backx F, et al. Prevalence of osteoarthritis in former elite athletes: a systematic overview of the recent literature[J]. Rheumatol Int, 2015, 35(3): 405-418.
[5] Vos T, Flaxman AD, Naghavi M, et al. Years lived with disability (YLDs) for 1160 sequelae of 289 diseases and injuries 1990-2010: a systematic analysis for the Global Burden of Disease Study 2010[J]. Lancet, 2014, 384(9943): 2163-2196.
[6] Pas HI, Winters M, Haisma HJ, et al. Stem cell injections in knee osteoarthritis: a systematic review of the literature[J]. Br J Sports Med, 2017, 3(0): 1-10.
[7] Li LU, Zhao Y. P16INK4a upregulation mediated by TBK1 induces retinal ganglion cell senescence in ischemic injury[J]. Cell Death Dis, 2017, 8(4): e2752.
[8] 郑文婕, 童坦君, 张宗玉. 细胞衰老的重要通路:p16INK4a/Rb和p19ARF/p53/p21Cip1信号途径[J]. 生命的化学, 2002, 22(4): 314-316.
[9] 扶世杰, 杨本伍, 舒从科, 等. 少阳生骨方对大鼠在体软骨损伤关节液IL-1β及软骨修复组织Ⅱ型胶原影响的实验研究[J]. 分子诊断与治疗杂志, 2014, 6(3): 191-197.
[10] Fabris L, Berton S, Pellizzari I, et al. p27kip1 controls H-Ras/MAPK activation and cell cycle entry via modulation of MT stability[J]. Proc Natl Acad Sci U S A, 2015, 112(45): 13916-13921.
[11] Quesnel B, Preudhomme C, Fenaux P. p16ink4a gene and hematological malignancies[J]. Leuk Lymphoma, 1996, 22(1-2):11-24.
[12] Ito K, Maruyama Z, Sakai A, et al. Overexpression of Cdk6 and Ccnd1 in chondrocytes inhibited chondrocyte maturation and caused p53-dependent apoptosis without enhancing proliferation[J]. Oncogene, 2014, 33(14):1862-1871.
[13] Li C, Chen L, Iwata T, et al. A Lys644Glu substitution in fibroblast growth factor receptor 3 (FGFR3) causes dwarfism in mice by activation of STATs and ink4 cell cycle inhibitors[J]. Hum Mol Genet, 1999, 8(1):35-44.
[14] Aszodi A, Hunziker EB, Brakebusch C,et al. Beta1 integrins regulate chondrocyte rotation, G1 progression, and cytokinesis[J]. Genes Dev, 2003, 17(19):2465.
[15] Huang Y, Wu G, Fan H, et al. Electroacupuncture promotes chondrocyte proliferation via accelerated G1/S transition in the cell cycle[J]. Int J Mol Med, 2013, 31(6):1443-1448.
[16] Ashraf S, Cha BH, Kim JS, et al. Regulation of senescence associated signaling mechanisms in chondrocytes for cartilage tissue regeneration[J]. Osteoarthritis cartilage, 2016, 24(2): 196-205.
[17] Yzydorczyk C, Li N, Chehade H, et al. Transient postnatal overfeeding causes liver stress-induced premature senescence in adult mice[J]. Sci Rep, 2017, 7(1): 12911.
[18] Wu G, Fan H, Huang Y, et al. Duhuo Jisheng Decoctioncontaining serum promotes proliferation of interleukin1βinduced chondrocytes through the p16cyclin D1/CDK4Rb pathway[J]. Mol Med Rep, 2014, 10(5): 2525-2534.

少阳主骨方对食蟹猴自然退变膝关节骨性关节炎模型p16/Rb信号通路的影响

Effects of ShaoYangZhuGuFang on p16/Rb signaling pathway of naturally degenerative cynomolgus monkeys knee osteoarthritis model

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