[1]李田花,施珊妮,桂苗,等.阳和汤治疗兔膝骨关节炎阳虚寒凝证的效果及作用机制[J].中医正骨,2023,35(12):14-19，24.
　LI Tianhua,SHI Shanni,GUI Miao,et al.Efficacy and mechanism of Yanghe Tang(阳和汤)for treatment of syndrome of Yang deficiency due to Yin excess in knee osteoarthritis rabbits[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2023,35(12):14-19，24.
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阳和汤治疗兔膝骨关节炎阳虚寒凝证的效果及作用机制()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第35卷
期数:
2023年12期
页码:
14-19，24
栏目:
基础研究
出版日期:
2023-12-20

文章信息/Info

Title:
Efficacy and mechanism of Yanghe Tang(阳和汤)for treatment of syndrome of Yang deficiency due to Yin excess in knee osteoarthritis rabbits
作者:
李田花1; 施珊妮1; 桂苗1; 黄泽灵2; 邱添华1; 高弘建1; 洪振强3
1.福建中医药大学中医学院,福建 福州 350122; 2.南京中医药大学附属苏州市中医医院,江苏 苏州 215009; 3.中医骨伤及运动康复教育部重点实验室,福建 福州 350122
Author(s):
LI Tianhua1; SHI Shanni1; GUI Miao1; HUANG Zeling2; QIU Tianhua1; GAO Hongjian1; HONG Zhenqiang3
1.School of Traditional Chinese Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian,China 2.Suzhou Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Chinese Medicine,Suzhou 215009,Jiangsu,China 3.Key Laboratory of Orthopedics&Traumatology and Rehabilitation of Traditional Chinese Medicine of Ministry of Education,Fuzhou 350122,Fujian,China
关键词:
骨关节炎; 膝;  寒凝阳虚;  阳和汤;  模型; 动物;  兔;  信号传导
Keywords:
osteoarthritis; knee;  Yang deficiency due to Yin excess;  Yang He Tang;  models; animal;  rabbits;  signal transduction
摘要:
目的:探讨阳和汤治疗膝骨关节炎(knee osteoarthritis,KOA)阳虚寒凝证的效果及作用机制。方法:将24只新西兰兔随机分为正常对照组和造模组,正常对照组6只,造模组18只。造模组兔采用双侧后肢膝关节腔内注射木瓜蛋白酶建立KOA模型,同时采用薄荷油灌胃联合4 ℃人工气候箱饲养建立阳虚寒凝证模型。造模成功后将18只KOA阳虚寒凝证兔随机分为模型组、阳和汤干预组和塞来昔布干预组,每组6只。阳和汤干预组兔按照中药饮片3 g·kg-1的剂量给予阳和汤浓缩剂灌胃,塞来昔布干预组按照0.02 g·kg-1的剂量给予塞来昔布溶液灌胃,模型组和正常对照组给予等量生理盐水灌胃。每日给药1次,连续给药2周。末次给药后当天,采用改良版Lequense MG评分标准评价兔膝关节疼痛及功能,采用桂苗等制定的阳虚证候评分表及寒凝证候评分表评价兔阳虚寒凝证候改善情况。末次给药后禁食不禁水24 h,处死兔并取材,采用HE染色观察膝关节滑膜组织病理改变,采用实时定量PCR检测膝关节滑膜组织中Toll样受体4(Toll-like receptor 4,TLR4)、核因子-κB(nuclear factor-κB,NF-κB)的mRNA表达量,采用蛋白免疫印迹法检测膝关节滑膜组织中TLR4、NF-κB的蛋白表达量,采用酶联免疫吸附法测定膝关节液中白细胞介素(interleukin,IL)-1β、IL-6、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的含量。结果:阳和汤干预组和塞来昔布干预组兔膝关节改良版Lequesne MG评分、阳虚寒凝证候评分均小于模型组(阳和汤干预组:P=0.000,P=0.000; 塞来昔布干预组:P=0.000,P=0.000); 阳和汤干预组兔膝关节改良版Lequesne MG评分与塞来昔布干预组的差异无统计学意义(P=0.751),阳虚寒凝证候评分小于塞来昔布干预组(P=0.000); 阳和汤干预组和塞来昔布干预组膝关节滑膜表面较为平整,衬里层细胞层数较模型组减少,细胞排列较为紧密; 衬里下层仍可见较多脂肪细胞分布,可见少量炎性细胞浸润,纤维组织、血管少量增生; 阳和汤干预组和塞来昔布干预组兔膝关节滑膜组织中TLR4和NF-κB的mRNA表达量及蛋白表达量均小于模型组(阳和汤干预组:P=0.000,P=0.000; P=0.000,P=0.000; 塞来昔布干预组:P=0.000,P=0.000; P=0.000,P=0.000); 阳和汤干预组和塞来昔布干预组兔膝关节液中TNF-α、IL-1β、IL-6的含量均小于模型组(阳和汤干预组:P=0.000,P=0.000,P=0.000; 塞来昔布干预组:P=0.000,P=0.000,P=0.000)。结论:阳和汤治疗KOA阳虚寒凝证,能够显著缓解膝关节疼痛,改善膝关节功能、阳虚寒凝证候及膝关节滑膜组织病理表现,在缓解膝关节疼痛和改善膝关节功能方面与塞来昔布相当,在改善阳虚寒凝证候方面较塞来昔布更具优势; 其作用机制可能是通过下调TLR4、NF-κB的表达,抑制TNF-α、IL-6及IL-1β等炎症因子的表达。
Abstract:
Objective:To explore the effects and mechanism of Yanghe Tang(阳和汤,YHT)in treatment of syndrome of Yang deficiency due to Yin excess in knee osteoarthritis(KOA)rabbits.Methods:Twenty-four New Zealand rabbits were randomized into normal control group(6 ones)and modeling group(18 ones).The rabbits in modeling group were intervened by knee intra-articular injection of papain into bilateral hind limbs for inducing KOA,followed by intragastric administration with peppermint oil and reared in a 4 ℃ artificial climate chamber for inducing the syndrome of Yang deficiency due to Yin excess.After successful modeling,the 18 modeled rabbits were randomized into model group,YHT intervention group,and celecoxib intervention group,6 cases in each group.The rabbits in YHT intervention group were intragastric administrated with YHT concentrate in dosage of 3 g/kg of traditional Chinese medicine decoction pieces,the ones in celecoxib intervention group with celecoxib solution in dosage of 0.02 g/kg,while the ones in model group and normal control group with the same dose of normal saline.All rabbits were intervened once a day for consecutive 2 weeks.On the day of the last intervention,the knee pain and function of the rabbits were evaluated by using the modified Lequesne MG knee assessment scale,and the syndrome alleviation was assessed by using the Yang deficiency and Yin excess scoring scales developed by Gui Miao et al.After the last intervention,the rabbits were deprived of food but water for 24 hours,then they were sacrificed and their knee synovial tissues were harvested and stained with hematoxylin-eosin(HE)for observing the pathological changes.Furthermore,the mRNA and protein expression levels of Toll-like receptor 4(TLR4)and nuclear factor-κB(NF-κB)in the knee synovial tissues were detected by using real-time quantitative PCR(RT-qPCR)and Western blotting, respectively,and the levels of interleukin(IL)-1β,IL-6,and tumor necrosis factor-α(TNF-α)in the knee synovial fluid were detected by using enzyme linked immunosorbent assay(ELISA).Results:The modified Lequesne MG score and syndrome(Yang deficiency due to Yin excess)score were lower in YHT intervention group and celecoxib intervention group compared with that in model group(YHT intervention group:P=0.000,P=0.000; celecoxib intervention group:P=0.000,P=0.000).The comparison of modified Lequesne MG score between YHT intervention group and celecoxib intervention group revealed no significant differences(P=0.751); while,the decreased syndrome score behaved in YHT intervention group compared to celecoxib intervention group(P=0.000).Compared with that in the model group,the knee synovial membrane in YHT intervention group and celecoxib intervention group showed smooth surface with reduced layers of lining cells,compact cell arrangement,existence of adipocytes among sublining cells,infiltration of a small amount of inflammatory cells,and mild proliferation of fibrous tissues and blood vessels.The YHT intervention group and celecoxib intervention group displayed down-regulated mRNA and protein expression levels of TLR4 and NF-κB in the knee synovial tissues of rabbits(YHT intervention group:P=0.000,P=0.000; P=0.000,P=0.000; celecoxib intervention group:P=0.000,P=0.000; P=0.000,P=0.000),and lowered levels of TNF-α,IL-1β,and IL-6 in the knee synovial fluid of rabbits compared to model group(YHT intervention group:P=0.000,P=0.000,P=0.000; celecoxib intervention group:P=0.000,P=0.000,P=0.000).Conclusion:YHT can obviously relieve the knee pain and improve the knee function,the syndrome of Yang deficiency due to Yin excess and the pathological manifestations of knee synovial tissues in treating the syndrome of Yang deficiency due to Yin excess of KOA.It is comparable to celeoxib in alleviating knee pain and improving knee function,while,it outperforms celecoxib in mitigating the syndrome of Yang deficiency due to Yin excess.It may exert the effects by inhibiting the expressions of TNF-α,IL-6 and IL-1β through down-regulating the expressions of TLR4 and NF-κB.

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备注/Memo

备注/Memo:
基金项目:福建中医药大学校管理科研项目(X2019006-学科)
通讯作者:洪振强 E-mail:594710961@qq.com
更新日期/Last Update: 1900-01-01