低频超声促透通络止痛方对大鼠不同程度膝骨关节炎的影响

Effects of Herbs Removing Obstruction in Channels to Relieve Pain Processed by Low-frequency Ultrasonic Transdermal Penetration on Rats with Different Degrees Knee Osteoarthritis

目的探讨通络止痛方在低频超声促透下对大鼠不同程度膝骨关节炎(Knee osteoarthritis,KOA)模型的行为学、软骨及滑膜组织形态学的影响,阐述该疗法的作用机制及其优势。方法采用切断前交叉韧带的经典方法制备KOA大鼠模型。SD大鼠随机分为3组:对照组、超声促透组和中药外敷组。所有大鼠左膝造模,对照组正常饲养,术后2、4、8周时各随机选取大鼠观察其行为学改变;处死后观察膝关节软骨大体改变,膝关节滑膜及股骨内侧髁软骨组织形态学改变,根据软骨改良Mankin’s评分予以评价,并根据评分结果确定大鼠KOA的病变程度。超声促透组和中药外敷组在术后2、4、8周时各随机选取大鼠进行干预处理,超声促透组使用超声电导仪对术侧膝关节促透通络止痛方,中药外敷组仅使用中药外敷。1次/日,30min,连续治疗2周。2周后处死,记录数据同对照组。结果对照组造模后2、4、8周时大鼠行为学、软骨大体观察、软骨及滑膜组织形态学发生明显改变,软骨Mankin’s评分的增加与时间成正相关。超声促透组与对照组相比,大鼠行为学发生改变,Lequesne MG评分在2、4、8周组时降低,有统计学差异(P<0.05);软骨组织形态学发生改变,Mankin’s评分2、4周组降低,2周时差异显著(P<0.01),4周时存在统计学差异(P<0.05);其作用显著优于中药外敷组,与中药外敷组相比,Lequesne MG评分在4周时下降明显,Mankin’s评分2周时下降明显,两两相比有统计学差异(P<0.05)。超声促透组与对照组和中药外敷组相比,滑膜组织形态学2、4、8周组发生改变,滑膜炎症反应降低;与对照组相比,软骨大体观察在2、4周组发生改变,退变程度减轻;与中药外敷组相比无明显改变。结论低频超声促透通络止痛方可以有效改善不同程度KOA的症状,减缓疾病的进一步发展,其机制可能是通过减缓软骨退变、抑制滑膜炎症而发挥作用,但对重度KOA软骨退变的缓解作用并不明显,与传统的中药外敷法相比,具有起效快,疗效好等优点。

OBJECTIVE To investigate the effects of herbs removing obstruction in channels to relieve pain processed by low-frequency ultrasonic transdermal penetration on rat models with different degrees knee osteoarthritis （KOA） in changes of behavior, cartilage and synovial histomorphology and explain its mechanisms and advantages of the therapy. METHODS The rat models of KOA were established by classic method of cutting off anterior cruciate ligaments. SD rats were randomly divided into three groups： blank control group, ultrasonic transdermal penetration group and external application of Chinese herbs group. All rats were modeled on left knees. The rats in blank control group were normally fed and randomly selected to observe their changes of behaviors at 2nd, 4th and 8th week after surgery. General changes of cartilage, synovium and medial femoral condylar cartilage histomorphology were observed and evaluated by cartilage modified Mankin＇s score to determine the lesion degree of KOA after rats in blank control group were sacrificed. The ultrasonic transdermal penetration group and external application of Chinese Herbs group were randomly selected for intervention treatment at 2nd, 4th and 8th week after surgery. The ultrasonic transdermal penetration group were treated with ultrasound conductimeter to promote herbs removing obstruction in channels to relieve pain on operating knees, while the external application of Chinese herbs group was just intervened by topical herbs, with one time per day for 2nd weeks. After 2 weeks, the rats were sacrificed, and the data was recorded to compare with the blank control group. RESULTS Rats in blank control group changed significantly in behavior, cartilage and synovial histomorphology at 2nd, 4th and 8th week after model establishment. The Mankin＇s score was positively related to time. Compared to the control group, behavioral changes in ultrasonic transdermal penetration group were more obvious, and Lequesne MG score decreased at 2nd, 4th and 8th week. There was statistical difference between them （P〈0.05）. Cartilage tissue morphology changed with decrease of Mankin＇s score at 2nd and 4th week, and there was significant difference at 2nd week （P〈0.01） and at 4th week （P 〈0.05）. It＇s more effeetive than external application of Chinese herbs group： Lequesne MG score decreased significantly at 4th week and Mankin＇s score decreased signifieantly at 2nd week. There was significant statistical difference （P 〈0.05）. Compared to blank control group and external application of Chinese Herbs group, ultrasonic transdermal penetration group showed more synovial tissue morphological changes and less inflammatory reactions of synovium at 2nd, 4th and 8th week. Cartilage changed and degenerated less at 2nd and 4th week when compared to blank control group, but there was no significant changes compared to external application of Chinese Herbs group, CONCLUSION Herbs removing obstruction in channels to relieve pain with low-frequency ultrasonic transdermal penetration can alleviate symptoms of different degrees of KOA and slow down the progression of the disease. The mechanisms may manifest as slowing down cartilage degeneration and inhibiting inflammations of synovium, but it is not effective in alleviating severe cartilage KOA. Removing obstruction in channels to relieve pain herbs with low-frequency ultrasonic transdermal penetration possesses better and quicker efficacy compared to traditional external application of Chinese herbs.