| 黄连素调控软骨下骨板骨保护素/核因子-κB受体活化剂配体系统延缓兔骨关节炎发展的实验研究 |
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Received:April 14, 2021
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| 作者 | Author | 单位 | Unit | E-Mail |
| 张建方 |
ZHANG Jian-fang |
浙江省立同德医院骨科, 浙江 杭州 310012 |
Department of Orthopaedics, Tongde Hospital of Zhejiang Province, Hangzhou 310012, Zhejiang, China |
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| 黄恺 |
HUANG Kai |
浙江省立同德医院骨科, 浙江 杭州 310012 |
Department of Orthopaedics, Tongde Hospital of Zhejiang Province, Hangzhou 310012, Zhejiang, China |
hzhuangk@163.com |
| 蔡海丽 |
CAI Hai-li |
中国人民解放军联勤保障部队第903医院特诊科, 浙江 杭州 310012 |
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| 张朋里 |
ZHANG Peng-li |
浙江省立同德医院骨科, 浙江 杭州 310012 |
Department of Orthopaedics, Tongde Hospital of Zhejiang Province, Hangzhou 310012, Zhejiang, China |
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| 王健 |
WANG Jian |
浙江省立同德医院骨科, 浙江 杭州 310012 |
Department of Orthopaedics, Tongde Hospital of Zhejiang Province, Hangzhou 310012, Zhejiang, China |
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| 期刊信息：《中国骨伤》2022年35卷，第5期，第464-469页 |
| DOI：10.12200/j.issn.1003-0034.2022.05.011 |
| 基金项目:浙江省中医药科技计划(编号:2018ZB033);国家自然科学基金面上项目(编号:81871792);浙江省医药卫生科技项目(编号:2020KY498) |
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目的:探讨关节腔内黄连素注射对兔骨关节炎(osteoarthritis,OA)软骨下骨板结构重塑以及骨保护素/核因子-κβ受体活化剂配体(osteoprotegerin/receptor activator of nuclear factor kappa-B ligand,OPG/RANKL)系统表达的影响。
方法:选取40 只12月龄雄性家兔,体重(2.73±0.18) kg,行左侧膝关节前十字韧带离断术(anterior cruciate ligament transection,ACLT),术后分为黄连素组和安慰剂组,每组20 只。黄连素组术后每周给予左膝关节腔内注射100 μmol/L 黄连素0.3 ml,共6周。安慰剂组术后每周给予左膝关节腔内注射相同剂量0.9%氯化钠注射液,共6周。另取12月龄雄性家兔20只,体重(2.68±0.18) kg,左膝关节行假手术,术后关节腔内不做注射干预(假手术组)。术后第6 周最后1 d,处死3组动物获取膝关节标本。获取股骨内髁标本行软骨及软骨下骨组织学评估,采用Mankin评分系统评估关节软骨结构,采用Image-Pro Plus(IPP)软件评估软骨下骨板骨小梁体积 (bone volume,BV),骨小梁体积百分比(bone volume/total volume,BV/TV),骨小梁周长(trabecular circumference,TC),平均骨小梁厚度(mean trabecular thickness,Tb.Th),采用实时定量反转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)检测术后6周软骨下骨组织中的OPG,RANKL的 mRNA 表达水平。
结果:软骨结构评估显示假手术组软骨组织表面光滑平整,番红在软骨全层着色丰满;黄连素组软骨组织可见表层不平整,番红素O染色轻度减退;安慰剂组可见表层纤维化,番红素O褪色明显。黄连素组Mankin评分低于安慰剂组(P<0.01),但高于假手术组(P<0.01)。软骨下骨板结构评估显示假手术组骨小梁排列致密;黄连素干预后小梁排列离较为紧密;安慰剂组软骨下骨小梁相对稀疏,小梁间距离较宽。软骨下骨板IPP软件评估显示,黄连素组BV、BV/TV、TC、Tb.Th 均高于安慰剂组(P<0.01),黄连素组BV、BV/TV、TC、Tb.Th 均低于假手术组(P<0.01)。PCR检测结果显示,黄连素组OPG mRNA表达明显高于安慰剂组(P<0.01),黄连素组OPG mRNA低于假手术组(P<0.05)。3组RANKL的 mRNA 表达比较差异无统计学意义(P>0.05);黄连素组OPG/RANKL 比值高于安慰剂组(P<0.01),低于假手术组(P<0.01)。
结论:关节腔内注射黄连素可以有效抑制 OA 早期软骨下骨吸收,延缓疾病发展,具体机制可能是黄连素通过上调软骨下骨 OPG 基因表达,维持了 OPG/RANKL 系统的平衡。 |
| [关键词]:骨关节炎 黄连素 软骨下骨重塑 OPG/RANKL系统 |
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| Experimental study on the regulation of subchondral bone plate osteoprotegerin/receptor activator of nuclear factor kappa-B ligand system by berberine to retard the development of osteoarthritis in rabbits |
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Abstract:
Objective: To investigate the effect of intra-articular berberine injection on the structural remodeling of subchondral bone plate and osteoprotegerin/receptor activator of nuclear factor kappa-B ligand(OPG/RANKL) system expression in rabbits with osteoarthritis(OA).
Methods: Forty 12-month-old male rabbits with an average of(2.73±0.18) kg of body weight,underwent left anterior cruciate ligament transection(ACLT),and were divided into berberine group and placebo groups after operation,20 rabbits in each group. The berberine group received intra-articular injection of 100 μmol/L berberine 0.3 ml every week for 6 weeks. In placebo group,the same dose of 0.9% sodium chloride injection was injected into the left knee joint cavity every week for 6 weeks. Another 20 12-month-old male rabbits,weighing (2.68±0.18) kg,underwent sham operation on the left knee joint without intra-articular injection intervention (sham operation group). On the last day of the sixth week after operation,three groups of animals were sacrificed to obtain knee joint specimens. The femoral medial condyle samples were obtained for histological evaluation of cartilage and subchondral bone,Mankin scoring system was used to evaluate articular cartilage structure. Image-Pro Plus(IPP) software was used to evaluate subchondral bone plate bone volume(BV), bone volume/total volume(BV/TV),trabecular circumference(TC),mean trabecular thickness (Tb.Th). Real-time quantitative reverse transcription polymerization Enzyme chain reaction(reverse transcription-polymerase chain reaction,RT-PCR) was used to detect the mRNA expression levels of OPG and RANKL in subchondral bone tissue at 6 weeks after operation.
Results: The cartilage structure evaluation showed that the surface of cartilage tissue in the sham operation group was smooth and flat,and the safranin coloration was full in the full thickness of the cartilage;the cartilage tissue in the berberine group showed uneven surface layer,and the staining of safranin O was mildly decreased;the surface layer fibrosis was seen in placebo group,Safranin O faded significantly. The Mankin score in the berberine group was lower than that in placebo group(P<0.01),but higher than that in sham operation group(P<0.01). The structural evaluation of subchondral bone plate showed that the trabecular bone in sham-operated group was densely arranged;after berberine intervention,the trabeculae were closely arranged;the subchondral bone trabeculae in placebo group were relatively sparse,and the distance between trabeculae was wider. Subchondral bone plate IPP software evaluation showed that BV,BV/TV,TC,Tb.Th in berberine group were higher than those in placebo group(P<0.01),BV,BV/TV,TC,Tb.Th in berberine group were higher than those in placebo group(P<0.01),while lower than the sham operation group (P<0.01). PCR test results showed that the expression of OPG mRNA in the berberine group was significantly higher than that in placebo group(P<0.01),and OPG mRNA in the berberine group was lower than that in sham operation group (P<0.01). There was no significant difference in mRNA expression of RANKL among three groups(P>0.05);the ratio of OPG/RANKL in berberine group was higher than that in placebo group(P<0.01),but lower than that in sham operation group(P<0.01).
Conclusion: Intra-articular injection of berberine can effectively inhibit the resorption of subchondral bone in the early stage of OA and delay the development of the disease. The specific mechanism may be that berberine maintains the balance of OPG/RANKL system by up-regulating the expression of OPG gene in subchondral bone. |
| KEYWORDS:Osteoarthritis Berberine Subchondral bone remodeling OPG/RANKL system |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 张建方,黄恺,蔡海丽,张朋里,王健.黄连素调控软骨下骨板骨保护素/核因子-κB受体活化剂配体系统延缓兔骨关节炎发展的实验研究[J].中国骨伤,2022,35(5):464~469 |
| 英文格式: | ZHANG Jian-fang,HUANG Kai,CAI Hai-li,ZHANG Peng-li,WANG Jian.Experimental study on the regulation of subchondral bone plate osteoprotegerin/receptor activator of nuclear factor kappa-B ligand system by berberine to retard the development of osteoarthritis in rabbits[J].zhongguo gu shang / China J Orthop Trauma ,2022,35(5):464~469 |
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