肾虚血瘀型SD大鼠膝骨关节炎模型的建立与验证  

Establishment and validation of the Sprague-Dawley rat model of osteoarthritis with kidney deficiency and blood stagnation

作　　者：杨程 李玉生 焦泓焯 尚曼[3] 刘琪 李林臻 樊方阳 张成龙 张晓宇[1,2] 张君涛 Yang Cheng;Li Yusheng;Jiao Hongzhuo;Shang Man;Liu Qi;Li Linzhen;Fan Fangyang;Zhang Chenglong;Zhang Xiaoyu;Zhang Juntao(The First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300381,China;National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300381,China;School of Basic Medical Sciences,Tianjin Medical University,Tianjin 300193,China)

机构地区：[1]天津中医药大学第一附属医院,天津市300381 [2]国家中医针灸临床医学研究中心,天津市300381 [3]天津医科大学基础医学院,天津市300193

出　　处：《中国组织工程研究》2024年第27期4273-4280,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(82074470),项目负责人:张君涛。

摘　　要：背景:肾虚血瘀证候作为膝骨关节炎常见中医证候,又是膝骨关节炎的基本病机,二者之间有着重要联系。但肾虚血瘀证候如何促进膝关节软骨损伤及其潜在机制并不明确,还需进一步研究。目的:探讨肾虚血瘀证候对SD大鼠膝骨关节炎进程的影响。方法:将16只SD大鼠随机分为模型观察组和对照组,每组8只。模型观察组采用卵巢摘除方法建立肾虚模型,2个月后采用改良HULTH手术进行膝骨关节炎造模,HULTH手术后1周皮下注射盐酸肾上腺素进行血瘀造模;对照组给予卵巢摘除假手术,改良HULTH手术,皮下注射生理盐水。HULTH手术后第5周,检测血液流变学、凝血四项、三碘甲状腺原氨酸、四碘甲状腺原氨酸及雌二醇水平;同时,拍摄膝关节X射线片,进行膝关节苏木精-伊红染色、番红-固绿染色和免疫组化染色。结果与结论:①与对照组相比,模型观察组全血黏度低切、中切、高切和血浆黏度显著升高,凝血四项中纤维蛋白原水平显著升高,凝血酶原时间和活化部分凝血活酶时间显著下降;三碘甲状腺原氨酸、四碘甲状腺原氨酸及雌二醇水平均显著下降;②影像学结果显示模型观察组大鼠膝关节间隙狭窄程度更严重且表面欠光滑,出现高密度影;③苏木精-伊红染色和番红-固绿染色表明,模型观察组大鼠软骨损伤更严重,并且OARSI评分和Mankin评分明显高于对照组;④免疫组化染色结果显示,相较于对照组,模型观察组大鼠软骨外基质Ⅱ型胶原、聚集蛋白聚糖蛋白表达显著下降,炎症因子基质金属蛋白酶13、解聚蛋白样金属蛋白酶5蛋白表达显著升高。结果表明,该研究成功建立了SD大鼠肾虚血瘀型膝骨关节炎模型;肾虚血瘀证候通过促进炎症因子表达进一步加重软骨外基质裂解和软骨退变,促进大鼠膝骨关节炎进程。BACKGROUND:Kidney deficiency and blood stasis syndrome are common traditional Chinese medicine syndromes observed in knee osteoarthritis,which serve as fundamental pathogenesis factors.There exists a significant connection between the two.Previous studies have demonstrated that kidney deficiency and blood stasis syndrome effectively contribute to knee joint cartilage degeneration and the progression of knee osteoarthritis.However,the mechanisms underlying the promotion of knee joint cartilage damage remain unclear and require further investigation.OBJECTIVE:To investigate the influence of kidney deficiency and blood stasis syndrome on the progression of knee osteoarthritis in Sprague-Dawley rats.METHODS:Sixteen Sprague-Dawley rats were randomly divided into two groups:a model observation group and a control group,with eight rats in each group.Animal models of kidney deficiency were induced by ovary removal in the model observation group,while the control group was given a sham procedure for ovarian removal.Two months after modeling,both groups underwent modified HULTH surgery to induce knee osteoarthritis.One week after modified HULTH surgery,the model observation group was subcutaneously given adrenaline hydrochloride to make blood stasis models,while the control group was subcutaneously given normal saline.At the 5th week after modified HULTH surgery,blood rheology,coagulation parameters,triiodothyronine,tetraiodothyronine,and estradiol levels were measured.Knee joint X-ray images were taken,and knee joint sections were stained with safranin O-fast green,hematoxylin-eosin,and immunohistochemistry.RESULTS AND CONCLUSION:Compared with the control group,the model observation group exhibited significant increases in whole blood viscosity at low,medium,and high shear rates,as well as increased plasma viscosity.Fibronectin levels in the coagulation parameters were significantly increased,while prothrombin time and activated partial thromboplastin time were significantly decreased.Triiodothyronine,tetraiodothyronine,a

关 键 词：膝骨关节炎 肾虚血瘀 卵巢摘除 改良HULTH术 肾上腺素 炎症 软骨退化 SD大鼠 

分 类 号：R459.9[医药卫生—治疗学] R363[医药卫生—临床医学] R274