摘要
目的:观察刮痧对膝关节骨关节炎(KOA)大鼠膝关节软骨组织炎性因子、细胞凋亡及自噬的影响,探讨刮痧治疗KOA的作用机制。方法:SD大鼠随机分为空白组、模型组和刮痧组,每组17只。采用右膝关节腔内注射谷氨酸钠碘乙酸复制KOA大鼠模型。刮痧组选择右侧“阳陵泉”“血海”进行刮痧治疗,频率为1次/s,按压力度在0.3~0.5 kgf,隔日干预1次,连续干预7次。记录各组大鼠右膝关节周径变化;HE染色法观察大鼠右膝关节软骨病理形态变化;ELISA法检测大鼠右膝关节软骨组织中白细胞介素(IL)-1β及肿瘤坏死因子(TNF)-α含量;荧光定量PCR法和Western blot法分别检测大鼠膝关节软骨组织中自噬性关键小分子酵母Atg6的同素系物(Beclin-1)、微管相关结合蛋白1轻链蛋白酶复合物3Ⅱ/Ⅰ型(LC3Ⅱ/LC3Ⅰ)、泛素相关结合蛋白62(P62)、半胱天冬氨酸蛋白酶-3(Caspase-3)mRNA和蛋白的表达水平;TUNEL染色观察大鼠膝关节软骨细胞的凋亡情况;免疫荧光法检测大鼠右膝关节软骨中LC3的表达水平。结果:造模后,与空白组比较,模型组右侧膝关节周径明显增大(P<0.01);软骨层有明显的紊乱和缺损,软骨细胞大量坏死,有纤维增生及大量骨小梁破坏;软骨组织细胞凋亡率、IL-1β和TNF-α含量及Caspase-3、P62 mRNA和蛋白表达水平均升高(P<0.01,P<0.001),Beclin-1、LC3 mRNA和蛋白表达水平和LC3平均荧光强度降低(P<0.01,P<0.001)。治疗后,与模型组比较,刮痧组右侧膝关节周径缩小(P<0.05);软骨层排列趋于正常,炎性细胞浸润程度较低;软骨组织细胞凋亡率、IL-1β和TNF-α含量降低(P<0.05),Caspase-3、P62mRNA和蛋白表达水平降低(P<0.01,P<0.05),Beclin-1、LC3 mRNA和蛋白表达水平及LC3平均荧光强度升高(P<0.05)。结论:刮痧能够改善KOA大鼠关节软骨损伤,抑制炎性因子过量释放,减少软骨细胞凋亡率。这可能与上调Beclin-1、LC3的表达,降低P62、Caspase-3的表达,促�
Objective To observe the effect of Guasha on inflammation factors,apoptosis and autophagy in the cartilage tissue of knee joint in rats with knee osteoarthritis(KOA),so as to explore its mechanisms underlying im provement of KOA.Methods A total of 51 male SD rats were randomized into three groups:blank control,KOA model and Guasha(n=17 in each group).The rats in the blank control group received intra-articular injection of 0.9% NaCl solution in the right knee joint.The KOA model was established by intraarticular injection of glutamate sodium iodoacetic acid in the right knee joint.For rats of the Guasha group,Guasha(at a frequency of 1 time/s,and an applied pressure of 0.3—0.5 kgf)was applied to“Yanglingquan”(GB34)and“Xuehai”(SP10)areas of the right leg,once every other day,for 7 consecutive sessions.The circumference of the right knee was measured,The histopathological changes of right knee cartilage were observed after H.E.staining.The contents of inflammatory factors interleukin(IL)-1β and tumor necrosis factor(TNF)-αin the right knee articular cartilage tissue were assayed using ELISA.The expression levels of autophagy-related key molecule Beclin-1(homologous series of yeast Atg6),light chain protease complication 3 type Ⅱ/Ⅰ(LC3II/LC3 I),ubiquitin binding factor 62(P62)and cysteine aspartate protease-3(Caspase-3)mRNAs and proteins of the right knee articular cartilage tissue were measured using real-time fluorescent quantitative PCR and Western blot,separately.The apoptosis of chondrocytes was assayed using TUNEL staining,and the immunoactivity of LC3 determined using immunofluorescence staining.Results After modeling,the right knee circumference of the model and Guasha groups was significantly increased compared with the blank control group(P<0.01),and after the intervention,the knee circumference of the Guasha group was markedly decreased in comparison with that of the model group(P<0.05).Results of H.E.staining showed obvious degeneration and defects in the cartilage tissue,necrosis of a la
作者
颜雪华
朱浩
陈帅
张改月
张豪斌
杨金生
王莹莹
YAN Xue-hua;ZHU Hao;CHEN Shuai;ZHANG Gai-yue;ZHANG Hao-bin;YANG Jin-sheng;WANG Ying-ying(School of Acupuncture-moxibustion and Tuina,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province,China;The Second Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,Shaanxi Province;Institute of Basic Theory for Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700;Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences,Beijing 100700)
出处
《针刺研究》
CAS
CSCD
北大核心
2024年第3期247-255,共9页
Acupuncture Research
基金
国家自然科学基金项目(No.82074563)
2022年国家中医药管理局青年岐黄学者培养项目。
