摘要
目的探讨骨灵膏对兔膝关节软骨缺损模型骨组织再生的作用及对软骨细胞凋亡的影响。方法30只雄性新西兰兔,随机分为对照组、模型组以及骨灵膏低、中、高剂量组,每组6只。采用改良型关节刻痕法建立膝关节软骨缺损模型,对照组未钻孔,骨灵膏低、中、高剂量组灌胃给予1.16 mL/kg(3.70 g/kg生药量)、3.47 mL/kg(11.10 g/kg生药量)、10.41 mL/kg(33.31 g/kg生药量)骨灵膏,对照组、模型组给予3.47 mL/kg生理盐水,1次/3 d,连续给药12周。采用Wakitani评分标准进行组织学评分,采用Masson染色法进行病理学评估,采用缺口末端标记(terminal transferase-mediated dUTP nick end labeling,TUNEL)法检测滑膜、软骨组织细胞凋亡情况,采用酶联免疫吸附法检测尿液中的吡啶酚(pyridinophenol,PYD)以及关节液中的一氧化氮(nitric oxide,NO)、超氧化物歧化酶(superoxide dismutase,SOD)、过氧化脂质(lipid peroxide,LPO)浓度水平。结果模型组给药后第12周伤口处凹凸不平,填平不同的剂量组软骨缺损区,其中低剂量组的表面粗糙,中剂量组表明欠光滑,高剂量组表面较光滑,关节软骨结构良好。与模型组相比,各剂量组软骨细胞排列较为规律,软骨细胞分泌的胶原纤维较多。低、中、高剂量组的兔膝关节软骨缺损处组织形态学评分以及滑膜、软骨组织细胞凋亡率高于对照组,低于模型组,随着剂量增加,组织形态学评分以及滑膜、软骨组织细胞凋亡率均呈降低趋势(P<0.05)。低、中、高剂量组的关节液NO、LPO以及尿液PYD水平高于对照组,低于模型组,随着剂量的上涨,表达水平方面则呈下降的趋势,差异有统计学意义(P<0.05);低、中、高剂量组的关节液SOD水平低于对照组,高于模型组,随着剂量的上涨,表达水平方面则呈上升的趋势,差异有统计学意义(P<0.05)。结论骨灵膏可改善膝关节软骨缺损模型兔的软骨损伤情况,加快软骨组织修复,其作用机制可能为通过调控氧化应激因子表达而抑制软骨细胞凋亡,以及促进软骨组织再生。
Objective To investigate the effect of Guling Cream(骨灵膏)on bone regeneration and chondrocyte apoptosis in rabbit knee cartilage defect model.Methods Thirty male rabbits(New Zealand)were randomly divided into model group,blank control group and Guling Cream low,medium and high dose groups,with 6 rabbits in each group.The model of knee cartilage defect was established by modified joint scoring method.The blank control group did not drill,and the Guling Cream low,medium and high dose groups were given Guling Cream with dose of 1.16 mL/kg(3.70 g/kg),3.47 mL/kg(11.10 g/kg),10.41 mL/kg(33.31 g/kg)by gavage,respectively.The the model group blank and control group were given 3.47 mL/kg normal saline,once every 3 days for 12 weeks.The Wakitani score was used for histological score.Masson staining was used for pathological evaluation,and terminal transferase-mediated dUTP nick end labeling(TUNEL)method was used to detect synovial and chondrocyte apoptosis.The levels of superoxide dismutase(SOD),nitric oxide(NO),lipid peroxide(LPO)in synovial fluid and pyridine phenol(PYD)in urine were detected by enzyme-linked immunosorbent assay.Results At the 12th week after administration,the wound area in the model group was uneven,and the cartilage defect area in each dose group was filled in.The surface of the low dose group was rough,while the medium dose group showed less smoothness.The surface of the high dose group was relatively smooth,and the joint cartilage structure was good.Compared with those in the model group,the arrangement of chondrocytes in each dose group was more regular,and chondrocytes secreted more collagen fibers.The histological scores and apoptosis rates of synovial and chondrocytes in the low,medium and high dose groups of were higher than those in the control group and lower than those in the model group.As the dose increased,the histological scores and apoptosis rates of synovial and chondrocytes showed a decreasing trend(P<0.05).The levels of joint fluid NO and LPO and urine PYD in the low,medium and high dose groups were higher than those in the control group,but lower than those in the model group.As the dose increased,the expression levels showed a descending trend,and the difference was statistically obvious(P<0.05).The SOD levels in the joint fluid of the low,medium and high dose groups were lower than those of the control group and higher than those of the model group.As the dose increased,the expression level showed an growing trend,and the difference was statistically obvious(P<0.05).Conclusion Guling Cream can improve the cartilage injury and accelerate the repair of cartilage tissue in rabbits with knee cartilage defect possibly by regulating the expressions of oxidative stress factors to inhibit chondrocyte apoptosis and promote cartilage tissue regeneration.
作者
胡珂琦
干开丰
张挺
夏臣杰
周珂
沈延东
HU Keqi;GAN Kaifeng;ZHANG Ting;XIA Chenjie;ZHOU Ke;SHEN Yandong(Ningbo Medical Center Lihuili Hospital,Ningbo 315000,Zhejiang,China)
出处
《中华中医药学刊》
北大核心
2025年第4期221-224,共4页
Chinese Archives of Traditional Chinese Medicine
基金
浙江省自然科学基金项目(LQ21H060002)
宁波市医疗卫生高端团队重大攻坚项目(2022020102)
宁波市自然科学基金项目(2022J251)。
关键词
骨灵膏
软骨细胞
凋亡
膝骨性关节炎
软骨缺损
Guling Cream(骨灵膏)
chondrocytes
apoptosis
knee osteoarthritis
cartilage defect
