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[1]李田花,施珊妮,桂苗,等.陽和湯治療兔膝骨關(guān)節(jié)炎陽虛寒凝證的效果及作用機制[J].中醫(yī)正骨,2023,35(12):14-19，24.
　LI Tianhua,SHI Shanni,GUI Miao,et al.Efficacy and mechanism of Yanghe Tang(陽和湯)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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陽和湯治療兔膝骨關(guān)節(jié)炎陽虛寒凝證的效果及作用機制()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第35卷
期數(shù):
2023年12期
頁碼:
14-19，24
欄目:
基礎(chǔ)研究
出版日期:
2023-12-20

文章信息/Info

Title:
Efficacy and mechanism of Yanghe Tang(陽和湯)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; 黃澤靈2; 邱添華1; 高弘建1; 洪振強3
1.福建中醫(yī)藥大學(xué)中醫(yī)學(xué)院,福建 福州 350122; 2.南京中醫(yī)藥大學(xué)附屬蘇州市中醫(yī)醫(yī)院,江蘇 蘇州 215009; 3.中醫(yī)骨傷及運動康復(fù)教育部重點實驗室,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
關(guān)鍵詞:
骨關(guān)節(jié)炎; 膝;  寒凝陽虛;  陽和湯;  模型; 動物;  兔;  信號傳導(dǎo)
Keywords:
osteoarthritis; knee;  Yang deficiency due to Yin excess;  Yang He Tang;  models; animal;  rabbits;  signal transduction
摘要:
目的:探討陽和湯治療膝骨關(guān)節(jié)炎(knee osteoarthritis,KOA)陽虛寒凝證的效果及作用機制。方法:將24只新西蘭兔隨機分為正常對照組和造模組,正常對照組6只,造模組18只。造模組兔采用雙側(cè)后肢膝關(guān)節(jié)腔內(nèi)注射木瓜蛋白酶建立KOA模型,同時采用薄荷油灌胃聯(lián)合4 ℃人工氣候箱飼養(yǎng)建立陽虛寒凝證模型。造模成功后將18只KOA陽虛寒凝證兔隨機分為模型組、陽和湯干預(yù)組和塞來昔布干預(yù)組,每組6只。陽和湯干預(yù)組兔按照中藥飲片3 g·kg-1的劑量給予陽和湯濃縮劑灌胃,塞來昔布干預(yù)組按照0.02 g·kg-1的劑量給予塞來昔布溶液灌胃,模型組和正常對照組給予等量生理鹽水灌胃。每日給藥1次,連續(xù)給藥2周。末次給藥后當天,采用改良版Lequense MG評分標準評價兔膝關(guān)節(jié)疼痛及功能,采用桂苗等制定的陽虛證候評分表及寒凝證候評分表評價兔陽虛寒凝證候改善情況。末次給藥后禁食不禁水24 h,處死兔并取材,采用HE染色觀察膝關(guān)節(jié)滑膜組織病理改變,采用實時定量PCR檢測膝關(guān)節(jié)滑膜組織中Toll樣受體4(Toll-like receptor 4,TLR4)、核因子-κB(nuclear factor-κB,NF-κB)的mRNA表達量,采用蛋白免疫印跡法檢測膝關(guān)節(jié)滑膜組織中TLR4、NF-κB的蛋白表達量,采用酶聯(lián)免疫吸附法測定膝關(guān)節(jié)液中白細胞介素(interleukin,IL)-1β、IL-6、腫瘤壞死因子-α(tumor necrosis factor-α,TNF-α)的含量。結(jié)果:陽和湯干預(yù)組和塞來昔布干預(yù)組兔膝關(guān)節(jié)改良版Lequesne MG評分、陽虛寒凝證候評分均小于模型組(陽和湯干預(yù)組:P=0.000,P=0.000; 塞來昔布干預(yù)組:P=0.000,P=0.000); 陽和湯干預(yù)組兔膝關(guān)節(jié)改良版Lequesne MG評分與塞來昔布干預(yù)組的差異無統(tǒng)計學(xué)意義(P=0.751),陽虛寒凝證候評分小于塞來昔布干預(yù)組(P=0.000); 陽和湯干預(yù)組和塞來昔布干預(yù)組膝關(guān)節(jié)滑膜表面較為平整,襯里層細胞層數(shù)較模型組減少,細胞排列較為緊密; 襯里下層仍可見較多脂肪細胞分布,可見少量炎性細胞浸潤,纖維組織、血管少量增生; 陽和湯干預(yù)組和塞來昔布干預(yù)組兔膝關(guān)節(jié)滑膜組織中TLR4和NF-κB的mRNA表達量及蛋白表達量均小于模型組(陽和湯干預(yù)組:P=0.000,P=0.000; P=0.000,P=0.000; 塞來昔布干預(yù)組:P=0.000,P=0.000; P=0.000,P=0.000); 陽和湯干預(yù)組和塞來昔布干預(yù)組兔膝關(guān)節(jié)液中TNF-α、IL-1β、IL-6的含量均小于模型組(陽和湯干預(yù)組:P=0.000,P=0.000,P=0.000; 塞來昔布干預(yù)組:P=0.000,P=0.000,P=0.000)。結(jié)論:陽和湯治療KOA陽虛寒凝證,能夠顯著緩解膝關(guān)節(jié)疼痛,改善膝關(guān)節(jié)功能、陽虛寒凝證候及膝關(guān)節(jié)滑膜組織病理表現(xiàn),在緩解膝關(guān)節(jié)疼痛和改善膝關(guān)節(jié)功能方面與塞來昔布相當,在改善陽虛寒凝證候方面較塞來昔布更具優(yōu)勢; 其作用機制可能是通過下調(diào)TLR4、NF-κB的表達,抑制TNF-α、IL-6及IL-1β等炎癥因子的表達。
Abstract:
Objective:To explore the effects and mechanism of Yanghe Tang(陽和湯,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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備注/Memo

備注/Memo:
基金項目:福建中醫(yī)藥大學(xué)校管理科研項目(X2019006-學(xué)科)
通訊作者:洪振強 E-mail:[email protected]
更新日期/Last Update: 1900-01-01