加味青娥丸治疗膝骨关节炎的作用机制研究

Study on the mechanism of action of Jiawei Qing'e Wan( 加味青娥丸) for the treatment of knee osteoarthritis

目的:探讨加味青娥丸治疗膝骨关节炎(knee osteoarthritis,KOA)的作用机制。方法:将符合要求的120例KOA患者随机分为加味青娥丸组和芍药丸组,每组60例;分别口服加味青娥丸和芍药丸,每次1丸,每天3次,连续服用12周。服药期间2组患者均进行患肢皮肤牵引及不负重功能锻炼。当患者关节疼痛不能缓解或加重,无法忍受时,给予塞来昔布胶囊,每次1粒,每天1次,疼痛控制后立即停止服用塞来昔布胶囊。分别于治疗前和治疗12周后测定2组患者的膝关节疼痛视觉模拟评分(visual analogue score,VAS)和西安大略和麦克马斯特大学(Western Ontario and Mc Master Universities,WOMAC)骨关节炎指数评分,并测定患者血清白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和一氧化氮(nitric oxide,NO)水平,以及外周血单核细胞(peripheral blood mononuclear cell,PBMC)基质金属蛋白酶-3 mRNA(matrix metalloproteinase-3 mRNA,MMP-3 mRNA)表达水平。结果:1膝关节疼痛VAS评分及WOMAC评分。治疗前2组患者膝关节疼痛VAS评分及WOMAC评分比较,组间差异均无统计学意义(t=0.626,P=0.553;t=0.856,P=0.394);治疗12周后芍药丸组膝关节疼痛VAS评分及WOMAC评分均高于加味青娥丸组(t=9.075,P=0.000;t=17.149,P=0.000)。治疗12周后加味青娥丸组膝关节疼痛VAS评分及WOMAC评分均较治疗前降低(t=10.392,P=0.000;t=19.075,P=0.000);芍药丸组膝关节疼痛VAS评分及WOMAC评分治疗前后比较,差异均无统计学意义(t=0.664,P=0.508;t=1.860,P=0.065)。2血清IL-1β水平。治疗前2组各级别患者血清IL-1β水平比较,差异无统计学意义(F=0.612,P=0.894)。治疗12周后加味青娥丸组患者血清IL-1β水平与治疗前相比,差异有统计学意义(F=16.986,P=0.000);Ⅰ、Ⅱ级患者血清IL-1β水平较治疗前降低(P=0.000;P=0.000),Ⅲ、Ⅳ级患者血清IL-1β水平与治疗前相比,差异均无统计学意义(P=0.075;P=0.161)。治疗12周后芍药丸组各级别患者血清IL-1β水平与治疗前相比,差异无统计学意义(F=0.651,P=0.885)。治疗12周后2组患者血清IL-1β水平比较,差异有统计学意义(F=3.881,P=0.044);加味青娥丸组Ⅰ、Ⅱ级患者血清IL-1β水平均低于芍药丸组(P=0.008;P=0.000);2组Ⅲ、Ⅳ级患者血清IL-1β水平比较,组间差异无统计学意义(P=0.342;P=0.444)。3血清TNF-α水平。治疗前2组各级别患者血清TNF-α水平比较,差异无统计学意义(F=1.447,P=0.695)。治疗12周后加味青娥丸组患者血清TNF-α水平与治疗前相比,差异有统计学意义(F=103.189,P=0.000);Ⅰ、Ⅱ级患者血清TNF-α水平较治疗前降低(P=0.000;P=0.000),Ⅲ、Ⅳ级患者血清TNF-α水平与治疗前相比,差异均无统计学意义(P=0.281;P=0.079)。治疗12周后芍药丸组各级别患者血清TNF-α水平与治疗前相比,差异无统计学意义(F=1.065,P=0.786)。治疗12周后2组患者血清TNF-α水平比较,差异有统计学意义(F=13.958,P=0.003);加味青娥丸组Ⅰ、Ⅱ、Ⅳ级患者血清TNF-α水平均低于芍药丸组(P=0.000;P=0.000;P=0.018);2组Ⅲ级患者血清TNF-α水平比较,差异无统计学意义(P=0.125)。4血清NO水平。治疗前2组各级别患者血清NO水平比较,差异无统计学意义(F=0.505,P=0.918)。治疗12周后加味青娥丸组患者血清NO水平与治疗前相比,差异有统计学意义(F=25.740,P=0.000);Ⅰ、Ⅱ级患者血清NO水平较治疗前降低(P=0.000;P=0.000),Ⅲ、Ⅳ级患者血清NO水平与治疗前相比,差异均无统计学意义(P=0.080;P=0.121)。治疗12周后芍药丸组各级别患者血清NO水平与治疗前相比,差异无统计学意义(F=0.427,P=0.935)。治疗12周后2组患者血清NO水平比较,差异有统计学意义(F=5.621,P=0.039);加味青娥丸组Ⅰ、Ⅱ级患者血清NO水平均低于芍药丸组(P=0.000;P=0.000);2组Ⅲ、Ⅳ级患者血清NO水平比较,组间差异无统计学意义(P=0.062;P=0.226)。5PBMC MMP-3 mRNA水平。治疗前及治疗12周后,2组各级别患者PBMC MMP-3 mRNA水平比较,组间差异均无统计学意义(F=0.002,P=0.999;F=0.033,P=0.998)。治疗12周后加味青娥丸组和芍药丸组各级别患者MMP-3 mRNA水平与治疗前相比,差异均无统计学意义(F=0.029,P=0.999;F=0.002,P=0.999)。结论:加味青娥丸治疗早中期KOA的机理之一可能是通过各种途径下调血清IL-1β、TNF-α及NO水平,从而抑制软骨细胞凋亡和软骨基质降解。

Objective：To explore the mechanism of action of Jiawei Qing’e Wan（加味青娥丸,JWQEW）for the treatment of knee os-teoarthritis（KOA）.Methods：One hundred and twenty patients with KOA were randomly divided into two groups,60 cases in each group. The patients were treated with JWQEW and Shaoyao Wan（芍药丸,SYW）respectively,one pill 3 times a day for consecutive 12 weeks.All cases received skin traction and non-weight-bearing functional exercise in affected limbs during the treatment,and those patients who suf-fered from unrelieved or aggravated knee pain were given Celecoxib capsules,1 pill once a day.Celecoxib capsules would be withdrew as long as the knee pain was controlled.The knee pain visual analogue score（VAS）and Western Ontario and McMaster Universities（WOM-AC）osteoarthritis index scores were evaluated before the treatment and after 12 -week treatment respectively.The serum level of IL -1β, TNF -αand NO and the expression of PBMC MMP -3 mRNA were also detected at the same time.Results：There were no statistical differences in knee pain VAS scores and WOMAC scores between the two groups before the treatment（t =0.626,P =0.553;t =0.856,P =0.394）.After 12-week treatment,the knee VAS scores and WOMAC scores were higher in SYW group compared to JWQEW group（t =9.075,P =0.000;t =17.149,P =0.000）.After 12-week treatment,the knee VAS scores and WOMAC scores decreased in JWQEW group （t =10.392,P =0.000;t =19.075,P =0.000）.There were no statistical differences between pretreatment and post-treatment in knee VAS scores and WOMAC scores in SYW group（t =0.664,P =0.508;t =1.860,P =0.065）.There was no statistical difference in the serum level of IL -1βbetween the 2 groups before the treatment（F =0.612,P =0.894）.After 12-week treatment,there was statistical difference in the serum level of IL -1βin JWQEW group between pretreatment and post-treatment（F =16.986,P =0.000）.The serum level of IL -1βdecreased after the treatment in gradeⅠandⅡcases（P =0.000,P =0.000）,while there was no statistical difference in serum level of IL -1βbetween pretreatment and post-treatment in gradeⅢandⅣcases（P =0.075,P =0.161）.After 12-week treatment,there was no sta-tistical difference in the serum level of IL -1βin SYW group between pretreatment and post-treatment（F =0.651,P =0.885）.There was statistical difference in the serum level of IL -1βbetween the 2 groups after the treatment（F =3.881,P =0.044）.The serum IL -1βlevel was lower in JWQEW group compared to SYW Group in gradeⅠandⅡcases（P =0.008,P =0.000）,while there was no statistical differ-ence in the serum IL -1βlevel in gradeⅢandⅣcases between the 2 groups（P =0.342,P =0.444）.There was no statistical difference in the serum level of TNF -αbetween the 2 groups before the treatment（F =1.447,P =0.695）.After 12 -week treatment,there was statisti-cal difference in the serum level of TNF -αin JWQEW group between pretreatment and post-treatment（F =103.189,P =0.000）.The ser-um level of TNF -αdecreased in gradeⅠandⅡcases after the treatment（P =0.000;P =0.000）,while no statistical difference was found in gradeⅢandⅣcases（P =0.281,P =0.079）.After 12-week treatment,there was no statistical difference in the serum level of TNF -αin SYW group between pretreatment and post -treatment（F =1.065,P =0.786）and there was statistical difference between the two groups （F =13.958,P =0.003）.The serum level of TNF -αwas lower in gradeⅠ,Ⅱ,andⅣcases in JWQEW group compared to SYW group （P =0.000,P =0.000,P =0.018）,while there was no statistical difference in gradeⅢcases between the 2 groups（P =0.125）.There was no statistical difference in the serum level of NO between the 2 groups before the treatment（F =0.505,P =0.918）.There was statistical difference in the serum level of NO in JWQEW group between pretreatment and post-treatment（F =25.740,P =0.000）.The serum level of NO decreased in gradeⅠandⅡcases after the treatment（P =0.000,P =0.000）,while there was no statistical difference between pretreat-ment and post-treatment in gradeⅢandⅣcases（P =0.080,P =0.121）.After 12-weeks treatment,there was no statistical difference be-tween pretreatment and post-treatment in serum NO level in all grades of cases in SYW group（F =0.427,P =0.935）.There was significant difference in the serum level of NO between the two groups after 12-week treatment（F =5.621,P =0.039）.The serum level of NO was lower in gradeⅠandⅡcases in JWQEW group compared to SYW group（P =0.000,P =0.000）,while there was no statistical difference in gradeⅢandⅣcases between the 2 groups（P =0.062,P =0.226）.There was no statistical difference in the expression level of PBMC MMP-3 mRNA between the two groups before the treatment and after 12-week treatment（F =0.002,P =0.999;F =0.033,P =0.998）.There was no statistical difference in the expression level of PBMC MMP -3 mRNA between pretreatment and post-treatment in both of the two groups（F =0.029,P =0.999;F =0.002,P =0.999）.Conclusion：By down-regulating the serum levels of IL -1β,TNF -αand NO through various pathways,JWQEW can inhibit cartilage cell apoptosis and cartilage matrix degradation,which may be one of the mechanisms of action for treatment of early-middle KOA.