推拿对运动疲劳大鼠膝关节软骨退变的干预研究

Effect of Massage on Knee Articular Cartilage Degeneration in Rats with Exercise Fatigue

目的 研究推拿对运动疲劳大鼠膝关节软骨退变的干预效果。方法 将38只雄性SD大鼠随机分成5组,分别为C组(空白组)、M3组(3 d模型组)、T3组(3 d推拿干预组)、M7组(7 d模型组)、T7组(7 d推拿干预组)。C组6只,其余各组每组8只。C组自然喂养,不进行干预;M组与T组均采用“运动疲劳造模方法”进行造模,T组于造模结束后采用揉法分别施术于双侧股四头肌及腓肠肌,并通过柔性纳米传感器量化手法力度在4~5 N范围内。主要观察指标:通过HE染色及电镜检测观察组织形态学变化,通过ELISA法检测血清中超氧化物歧化酶(SOD)、丙二醛(MDA)的活性及含量。多样本均数间比较采用One-Way ANOVA检验,方差齐则采用LSD检验,方差不齐则采用Tamhane's T2检验。结果 1.形态学观察结果:(1)组织切片及HE染色:①C组:未见明显病理改变;②M3组:关节软骨结构不完整,局部关节软骨表层粗糙改变,浅层软骨细胞排列轻度紊乱,关节软骨表层有缺损表现,其他未见明显病理改变;③M7组:关节软骨结构不完整,局部关节软骨表层变得粗糙不整,浅层软骨细胞排列紊乱明显,软骨层变薄,软骨细胞坏死表现,关节软骨表层出现裂隙,其他未见明显病理改变;④T3组:关节软骨结构较完整,软骨细胞形态良好,软骨四层结构清晰且排列有序,未见明显表层软骨损伤及其他未见明显病理改变;⑤T7组:关节软骨基质着色均匀,结构较完整,四层结构较清晰,局部关节软骨表层不整,轻微缺损表现,其他未见明显病理改变。(2)电镜检测:①C组:软骨细胞呈卵圆形,且结构良好,细胞及胞膜完整,可观察到胞膜表面存在突起的微绒毛;细胞核及核膜结构完整,胞质内可观察到完整的线粒体、粗面内质网及高尔基复合体等细胞器,纤维组织排列整齐有序;②M3组:大部分软骨细胞发生肿胀,呈异常形态,胞膜上的微绒毛断裂,胞浆出现空泡化现象,细胞核形态异常,可见粗面内质网轻度扩张,腔隙增宽,线粒体肿胀,游离核糖体增多;③M7组:细胞形态异常明显,细胞膜边界模糊不清,胞浆内空泡化严重,细胞核形态不规则,核膜模糊,粗面内质网扩张明显,腔隙增宽,线粒体肿胀明显;④T3组:部分软骨细胞轻度肿胀,胞膜表面的微绒毛变短,胞浆存在空泡化现象,细胞核形态异常,粗面内质网轻度扩张,线粒体轻度肿胀;⑤T7组:可观察到部分软骨细胞肿胀明显,细胞膜边界破坏,胞膜表面的微绒毛变短,胞浆内空泡化明显,细胞核形态异常,粗面内质网轻度扩张,线粒体轻度肿胀2.ELISA检测结果:与K组相比,M3、M7组大鼠血清中SOD活力明显升高,差异极显著(P<0.01);与K组相比,M3组大鼠血清中MDA含量升高,差异显著(P<0.05),M7组大鼠血清中MDA含量明显升高,差异极显著(P<0.01)。结论 推拿疗法对运动疲劳大鼠膝关节软骨退变具有一定的干预效果。

Objective To study the effect of massage on knee cartilage degeneration in rats with exercise fatigue.Methods Thirty-eight male SD rats were randomly divided into five groups,which were Group C(blank group),Group M3(3-day model group),Group T3(3-day massage intervention group),Group M7(7-day model group),and Group T7(7-day massage intervention group).There were 6 animals in group C and 8 animals in each group in the remaining groups.Group C was fed naturally without intervention;group T was modeled using the“exercise fatigue modeling method”.Group T was treated with kneading method on the quadriceps femoris and gastrocnemius muscles,respectively,after the end of modeling,and the manual force was within the range of 4–5N by flexible nanosensors.MAIN OUTCOME MEASURES:Histomorphological changes were observed by HE staining and electron microscopy,and the activities and contents of superoxide dismutase(SOD)and malondialdehyde(MDA)in serum were detected by ELISA.One-Way ANOVA test was used to compare the means of multiple samples,LSD test was used for equal variance,and Tamhane’s T2 test was used for unequal variance.Results 1.Morphological observation results:(1)Histological sections and HE staining:①Group C:no obvious pathological changes were observed;②Group M3:the structure of articular cartilage was incomplete,the surface layer of local articular cartilage was rough and changed,the arrangement of superficial chondrocytes was slightly disordered,there were defects in the surface layer of articular cartilage,and others did not show obvious pathological changes;③Group M7:the structure of articular cartilage was incomplete,the surface layer of local articular cartilage became rough and uneven,the arrangement of superficial chondrocytes was obvious,the cartilage layer became thinner,chondrocyte necrosis showed,there were fissures in the surface layer of articular cartilage,and others did not show obvious pathological changes;④Group T3:the structure of articular cartilage was more complete,the shape of chondrocytes was good,the structure of four layers of cartilage was clear and orderly,no obvious pathological changes were observed;⑤Group T7:the staining of articular cartilage matrix was uniform,the structure was complete,the structure of four layers was clear,the surface layer of local articular cartilage was uneven,and there were slight defects.(2)Electron microscopy:①Group C:chondrocytes were oval,and the structure was good,the cells and membranes were intact,and protruding microvilli could be observed on the membrane surface;the structure of nucleus and nuclear membrane was intact,intact mitochondria,rough endoplasmic reticulum and Golgi complex and other organelles could be observed in the cytoplasm,and the fibrous tissue arrangement was orderly;②Group M3:most chondrocytes were swollen,showing abnormal morphology,microvilli on the membrane were broken,vacuolization phenomenon appeared in the cytoplasm,abnormal nuclear morphology,mild expansion of rough endoplasmic reticulum,widened lacunae,mitochondrial swelling,and increased free ribosomes were observed;③Group M7:abnormal cell morphology,blurred cell membrane boundary,severe vacuolization in the cytoplasm,irregular nuclear membrane morphology,blurred nuclear membrane,rough endoplasmic reticulum expansion,widened lacunae,and significant mitochondrial swelling;Mild mitochondrial swelling;④Group T3:some chondrocytes were slightly swollen,microvilli on the membrane surface were shortened,cytoplasm was slightly vacuolated,nuclear morphology was slightly abnormal,rough endoplasmic reticulum was slightly dilated,and mitochondria were slightly swollen;⑤Group T7:some chondrocytes could be observed to have significant swelling,the cell membrane boundary was destroyed,the microvilli on the cell membrane surface were shortened,there was vacuolization in the cytoplasm,the rough endoplasmic reticulum was slightly dilated,and the mitochondria were slightly swollen.2.ELISA test results:Compared with Group C,the SOD activity in the serum of rats in groups M3 and M7 was significantly increased,and the difference was extremely significant(P<0.01);compared with Group C,the MDA content in the serum of rats in group M3 was increased,the difference was significant(P<0.05);and the MDA content in the serum of rats in group M7 was significantly increased,and the difference was extremely significant(P<0.01).Conclusion Massage therapy has a certain intervention effect on knee cartilage degeneration in rats with exercise fatigue.