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[1]蔡宏杰,王旭,徐建波,等.小鼠膝骨關(guān)節(jié)炎陽虛血瘀證模型的構(gòu)建與評(píng)價(jià)[J].中醫(yī)正骨,2023,35(06):18-23.
　CAI Hongjie,WANG Xu,XU Jianbo,et al.Construction and evaluation of a mouse model of knee osteoarthritis with yang-deficiency and blood-stasis syndrome[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2023,35(06):18-23.
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小鼠膝骨關(guān)節(jié)炎陽虛血瘀證模型的構(gòu)建與評(píng)價(jià)()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第35卷
期數(shù):
2023年06期
頁碼:
18-23
欄目:
基礎(chǔ)研究
出版日期:
2023-06-20

文章信息/Info

Title:
Construction and evaluation of a mouse model of knee osteoarthritis with yang-deficiency and blood-stasis syndrome
作者:
蔡宏杰1; 王旭2; 徐建波1; 林士能2; 童培建1; 金紅婷1; 陳佳麗1
(1.浙江省中醫(yī)院,浙江 杭州 310006; 2.浙江中醫(yī)藥大學(xué)第一臨床醫(yī)學(xué)院,浙江 杭州 310053)
Author(s):
CAI Hongjie1; WANG Xu2; XU Jianbo1; LIN Shineng2; TONG Peijian1; JIN Hongting1; CHEN Jiali1
1.Zhejiang Provincial Hospital of Traditional Chinese Medicine,Hangzhou 310006,Zhejiang,China 2.The First Clinical Medical College of Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China
關(guān)鍵詞:
骨關(guān)節(jié)炎; 膝;  小鼠;  疾病模型; 動(dòng)物;  陽虛血瘀;  動(dòng)物實(shí)驗(yàn)
Keywords:
osteoarthritis; knee;  mice;  disease models; animal;  syndrome of yang deficiency and blood stasis;  animal experimentation
摘要:
目的:探討構(gòu)建小鼠膝骨關(guān)節(jié)炎(knee osteoarthritis,KOA)陽虛血瘀證模型的有效方法。方法:將12只10周齡雄性無特定病原C57BL/6J小鼠采用內(nèi)側(cè)半月板失穩(wěn)(destabilized medial meniscus,DMM)法構(gòu)建KOA模型。KOA造模成功后,將12只KOA模型小鼠隨機(jī)分為KOA陽虛血瘀證組和KOA對照組,每組6只。KOA陽虛血瘀證組小鼠于浸在冰水混合物中的塑料鼠箱中飼養(yǎng),并以冰水混合物為飲用水; KOA對照組小鼠于22 ℃干燥環(huán)境中正常飼養(yǎng)。飼養(yǎng)10周后,評(píng)價(jià)小鼠陽虛血瘀證表現(xiàn),分析小鼠步態(tài),處死小鼠后進(jìn)行小鼠膝關(guān)節(jié)Micro-CT分析、膝關(guān)節(jié)組織病理學(xué)觀察及膝關(guān)節(jié)軟骨Ⅱ型膠原蛋白α1鏈表達(dá)檢測。結(jié)果:①小鼠陽虛血瘀證評(píng)價(jià)結(jié)果。KOA陽虛血瘀證組小鼠的陽虛證評(píng)分和血瘀證評(píng)分均高于KOA對照組[(13.33±1.37)分,(3.50±1.23)分,t=13.120,P=0.000;(4.83±0.75)分,(0.83±0.75)分,t=9.200,P=0.000]。②小鼠步態(tài)分析結(jié)果。2組各收集到5只小鼠完整的步態(tài)分析數(shù)據(jù)。KOA陽虛血瘀證組小鼠右后肢的爪面積和步幅均小于KOA對照組[(0.610±0.094)cm2,(0.758±0.042)cm2,t=-3.211,P=0.012;(1.780±0.045)cm,(2.040±0.152)cm,t=-3.677,P=0.006]; 2組小鼠右后肢的擺動(dòng)期時(shí)間和站立期時(shí)間比較,組間差異均無統(tǒng)計(jì)學(xué)意義[(0.060±0.004)s,(0.060±0.009)s,t=-0.096,P=0.963;(0.062±0.009)s,(0.076±0.014)s,t=-1.810,P=0.137]。③小鼠膝關(guān)節(jié)Micro-CT分析結(jié)果。KOA陽虛血瘀證組小鼠的骨體積分?jǐn)?shù)低于KOA對照組[(55.582±2.810)%,(69.248±1.884)%,t=-6.997,P=0.002],骨小梁分離度大于KOA對照組[(0.114±0.009)mm,(0.077±0.010)mm,t=4.883,P=0.008]; 2組小鼠的骨小梁厚度、骨小梁數(shù)量比較,組間差異均無統(tǒng)計(jì)學(xué)意義[(0.148±0.005)mm,(0.210±0.036)mm,t=-2.939,P=0.094;(3.767±0.100)個(gè)·mm-1,(3.368±0.547)個(gè)·mm-1,t=1.243,P=0.282]。④小鼠膝關(guān)節(jié)組織病理學(xué)觀察結(jié)果。2組小鼠膝關(guān)節(jié)軟骨均有缺失,且KOA陽虛血瘀證組小鼠膝關(guān)節(jié)軟骨缺失較KOA對照組更嚴(yán)重。⑤小鼠膝關(guān)節(jié)軟骨Ⅱ型膠原蛋白α1鏈表達(dá)檢測結(jié)果。KOA陽虛血瘀證組小鼠膝關(guān)節(jié)軟骨Ⅱ型膠原蛋白α1鏈陽性表達(dá)面積比和平均光密度均低于KOA對照組[(0.144±0.003)%,(0.163±0.003)%,t=-6.580,P=0.003;(0.051±0.002)lux,(0.095±0.004)lux,t=-15.878,P=0.001]。結(jié)論:DMM法聯(lián)合低溫誘導(dǎo)法是構(gòu)建小鼠KOA陽虛血瘀證模型的有效方法。
Abstract:
Objective:To explore the effective method for building a yang-deficiency-blood-stasis-type knee osteoarthritis(KOA)mouse model.Methods:Twelve 10-week-old specific pathogen free(SPF)-grade male C57BL/6J mice were selected out and were intervened by destabilized medial meniscus(DMM)method for inducing KOA.After successful modeling,the 12 KOA model mice were randomly divided into yang-deficiency-blood-stasis-type KOA group and KOA control group,6 cases in each group.The mice in yang-deficiency-blood-stasis-type KOA group were bred in plastic box submerged in ice-water mixture,and they were fed with ice-water mixture,while the ones in KOA control group were bred normally in a dry environment of 22 ℃.After 10-week feeding,the yang-deficiency and blood-stasis syndrome in KOA model mice were observed and evaluated,and the gaits were analyzed.After that,the mice were executed by using cervical dislocation method,and the knee samples were harvested from their right hind limbs.The knee samples were scanned and analyzed by using micro-CT and CTAn1.10 software respectively,followed by observation on histopathological changes of knee tissues and determination of the expression levels of typeⅡcollagen α1 chain(Col2A1)in knee cartilage.Results:①The scores of yang-deficiency syndrome and blood-stasis syndrome were higher in yang-deficiency-blood-stasis-type KOA group compared to KOA control group(13.33±1.37 vs 3.50±1.23 points,t=13.120,P=0.000; 4.83±0.75 vs 0.83±0.75 points,t=9.200,P=0.000).②The complete gait analysis data was collected from 5 mice in each group.The paw area and stride length of the right hind limbs were smaller in yang-deficiency-blood-stasis-type KOA group compared to KOA control group(0.610±0.094 vs 0.758±0.042 cm(2),t=-3.211,P=0.012; 1.780±0.045 vs 2.040±0.152 cm,t=-3.677,P=0.006).There was no statistical difference in the time of swing phase and stance phase of the right hind limbs between the 2 groups(0.060±0.004 vs 0.060±0.009 seconds,t=-0.096,P=0.963; 0.062±0.009 vs 0.076±0.014 seconds,t=-1.810,P=0.137).③The bone volume fraction was lower and the trabecular separation was greater in yang-deficiency-blood-stasis-type KOA group compared to KOA control group(55.582±2.810 vs 69.248±1.884%,t=-6.997,P=0.002; 0.114±0.009 vs 0.077±0.010 mm,t=4.883,P=0.008); while there was no statistical difference in trabecular thickness and trabecular number between the 2 groups(0.148±0.005 vs 0.210±0.036 mm,t=-2.939,P=0.094; 3.767±0.100 vs 3.368±0.547 trabeculae/mm,t=1.243,P=0.282).④The knee cartilage loss was observed in the model mice of the 2 groups,and it was more severe in yang-deficiency-blood-stasis-type KOA group compared to KOA control group.⑤The positive expression area ratio and average optical density of Col2A1 in knee cartilage were lower in yang-deficiency-blood-stasis-type KOA group compared to KOA control group(0.144±0.003 vs 0.163±0.003%,t=-6.580,P=0.003; 0.051±0.002 vs 0.095±0.004 lux,t=-15.878,P=0.001).Conclusion:Combination of DMM with low temperature induction is an effective method for constructing a mouse model of KOA and yang-deficiency and blood-stasis syndrome.

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中醫(yī)正骨2023年6月第35卷第6期 J Trad Chin Orthop Trauma,2023,Vol.35,No.6(總423)
(總424)中醫(yī)正骨2023年6月第35卷第6期 J Trad Chin Orthop Trauma,2023,Vol.35,No.6
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備注/Memo

備注/Memo:
基金項(xiàng)目:國家自然科學(xué)基金項(xiàng)目(81873324); 浙江中醫(yī)藥大學(xué)基本科研能力提升項(xiàng)目(2021JKJNTZ014A) 通訊作者:陳佳麗 E-mail:[email protected]
更新日期/Last Update: 1900-01-01