通络止痛凝胶制剂对膝骨关节炎大鼠的影响
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摘要
目的:观察通络止痛凝胶制剂对膝骨关节炎(KOA)模型大鼠患膝直径、软骨形态及白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、基质金属蛋白酶-13(MMP-13)的影响,探讨通络止痛凝胶制剂治疗KOA的疗效及机制。方法:将48只Wistar大鼠随机分为正常组、模型组、中药组、西药组,每组12只。正常组不造模,其他3组采用改良Hulth法建立KOA模型。造模成功后,中药组、西药组分别外涂中药凝胶剂、扶他林乳膏,正常组、模型组分别涂以等剂量0.9%氯化钠溶液,3次/d,共2周。使用游标卡尺于不同时间点测量患膝直径,治疗结束后软骨组织切片采用番红-固绿染色后光镜下观察软骨组织形态,血清采用ELISA法、滑膜组织采用Western Blot法检测IL-1β、TNF-α、MMP-13的含量。结果:与模型组比较,治疗1周后中药组、西药组的膝关节直径变小(P<0.05,P<0.01),治疗2周后中药组优于西药组(P<0.05);与模型组比较,中药组、西药组关节软骨表面较平整,结构层次较清晰,中药组稍优于西药组;与模型组比较,中药组、西药组血清和滑膜组织中IL-1β、TNF-α、MMP-13的含量均显著降低(P<0.01,P<0.05);与西药组比较,中药组血清和滑膜组织中IL-1β、MMP-13的含量显著降低(P<0.05)。结论:通络止痛凝胶制剂治疗大鼠KOA安全有效,可以改善其软骨损伤并缓解滑膜炎症反应。
Objective: To observe the effects of Tongluo Zhitong Gel Preparation on knee diameter, cartilage morphology,interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α) and matrix metalloproteinase-13(MMP-13), and mechanism of Tongluo Zhitong Gel Preparation in the treatment knee osteoarthritis(KOA). Methods: Forty-eight Wistar rats were randomly divided into normal group, model group, Chinese medicine group and western medicine group, 12 rats in each group. The normal group did not be modeled, and the other three groups used the modified Hulth method to establish the KOA model. After successful modeling,the traditional Chinese medicine group and the western medicine group were respectively coated with traditional Chinese medicine gel and Futalin Cream. The normal group and the model group were respectively coated with an equal dose of 0.9% normal saline,they were treated 3 times/d for 2 weeks. The diameter of the knee was measured at different time points using a vernier caliper.After the treatment, the cartilage tissue sections were stained with saffron solid-green and observed under light microscope. The serum was detected by ELISA and the content of TNF-α, MMP-13 in synovial tissue was detected by Western Blot. Results:Compared with the model group, after 1 week treatment the knee joint diameter of the Chinese medicine group and the western medicine group became smaller(P<0.05, P<0.01), and after 2 week treatment the Chinese medicine group was superior to the western medicine group(P<0.05). Compared with the model group, the Chinese medicine group and the western medicine group had articular cartilage, and the surface was relatively flat and the structure level was relatively clear. The Chinese medicine group was slightly better than the western medicine group. Compared with the model group, the contents of IL-1β, TNF-α and MMP-13 in the serum and synovial tissues of the Chinese medicine group and the western medicine group were significantly lower(P<0.01,P<0.05); Compared with the western medicine group, the levels of IL-1β and MMP-13 in the serum and synovial tissues of the Chinese medicine group were significantly lower(P<0.05). Conclusion: Tongluo Zhitong Gel Preparation is safe and effective in treating rat KOA, which can improve cartilage damage and relieve synovial inflammation.
Objective: To observe the effects of Tongluo Zhitong Gel Preparation on knee diameter, cartilage morphology,interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α) and matrix metalloproteinase-13(MMP-13), and mechanism of Tongluo Zhitong Gel Preparation in the treatment knee osteoarthritis(KOA). Methods: Forty-eight Wistar rats were randomly divided into normal group, model group, Chinese medicine group and western medicine group, 12 rats in each group. The normal group did not be modeled, and the other three groups used the modified Hulth method to establish the KOA model. After successful modeling,the traditional Chinese medicine group and the western medicine group were respectively coated with traditional Chinese medicine gel and Futalin Cream. The normal group and the model group were respectively coated with an equal dose of 0.9% normal saline,they were treated 3 times/d for 2 weeks. The diameter of the knee was measured at different time points using a vernier caliper.After the treatment, the cartilage tissue sections were stained with saffron solid-green and observed under light microscope. The serum was detected by ELISA and the content of TNF-α, MMP-13 in synovial tissue was detected by Western Blot. Results:Compared with the model group, after 1 week treatment the knee joint diameter of the Chinese medicine group and the western medicine group became smaller(P<0.05, P<0.01), and after 2 week treatment the Chinese medicine group was superior to the western medicine group(P<0.05). Compared with the model group, the Chinese medicine group and the western medicine group had articular cartilage, and the surface was relatively flat and the structure level was relatively clear. The Chinese medicine group was slightly better than the western medicine group. Compared with the model group, the contents of IL-1β, TNF-α and MMP-13 in the serum and synovial tissues of the Chinese medicine group and the western medicine group were significantly lower(P<0.01,P<0.05); Compared with the western medicine group, the levels of IL-1β and MMP-13 in the serum and synovial tissues of the Chinese medicine group were significantly lower(P<0.05). Conclusion: Tongluo Zhitong Gel Preparation is safe and effective in treating rat KOA, which can improve cartilage damage and relieve synovial inflammation.
引文
[1]Wang K,Xu J,Cai J,et al.Serum levels of interleukin-17 and adiponectin are associated with infrapatellar fat pad volume and signal intensity alteration in patients with knee osteoarthritis.Arthritis Research&Therapy,2016,18(1):S369
[2]Mc Alindon T E,Annuru R R,Sullivan M C,et al.Oarsi guidelines for the non-surgical management of knee osteoarthritis.Osteoarthritis&Cartilage,2014,22:363-388
[3]Ondrésik M,Maia F R A,Morais A D S,et al.Management of knee osteoarthritis.Current status and future trends.Biotechnology&Bioengineering,2017,114(4):717-739
[4]Murphy N J,Eyles J P,Hunter D J.Hip Osteoarthritis:Etiopathogenesis and implications for management.Advances in Therapy,2016,33(11):1921-1946
[5]王庆甫,祁印泽,陈兆军,等.超声促透透射通络止痛中药外治膝骨性关节炎的临床观察.北京中医药大学学报,2010,33(4):283-285
[6]王庆甫,马玉峰,殷岳衫,等.低频超声促透中药对兔膝关节炎细胞因子的影响.北京中医药大学学报,2013,36(2):108-112
[7]杨黎黎,王欢,张栋,等.通络止痛方对人膝骨关节炎滑膜炎性细胞TLR4、IL-1β及NO的影响研究.中华中医药杂志,2017,32(9):4124-4226
[8]杨黎黎,王欢,张栋,等.通络止痛方对人膝骨关节炎滑膜炎症细胞上清IL-1β和TNF-α的影响.辽宁中医药杂志,2017,44(5):1040-1042
[9]Zhou X,Zhang L,Guo X,et al.A Macaca fascicularis knee osteoarthritis model developed by modified Hulth combined with Joint Scratches.Medical Science Monitor International Medical Journal of Experimental&Clinical Research,2018,24:3393-3404
[10]贺石林,王健,王净净.中医科研设计与统计学.长沙:湖南科学技术出版社,2006:48-49
[11]Nirmal P,Koppikar S,Bhondave P,et al.Influence of six medicinal herbs on collagenase-induced osteoarthritis in rats.Am J Chin Med,2013,41(6):1407-1425
[12]邓莉,王海波.膝骨性关节炎动物模型的研究进展.广东医学,2018,39(12):1895-1897
[13]张君涛,吴超超,张栋林,等.膝关节骨性关节炎动物模型研究进展.重庆医学,2018,47(15):2061-2063,2067
[14]章海凤,陈树涛,冒姣娜,等.热敏灸对膝骨性关节炎兔模型IL-1β、TNF-α、MMP-13的影响.中华中医药杂志,2018,33(9):3913-3917
[15]Qinglu Luo,Xi Qin,Yaxian Qiu,et al.The change of synovial fluid proteome in rabbit surgery-induced model of knee osteoarthritis.Am J Transl Res,2018,10(7):2087-2101
[16]肖春苟,邹锦慧,李力强,等.以改良Hulth法建立兔膝骨性关节炎关节软骨的病理变化.中国中医骨伤科杂志,2014,2(12):1-3
[17]崔和平,郭兴凤.多不饱和脂肪酸对人体神经系统保健作用研究进展.河南工业大学学报(自然科学版),2012,33(3):97-102
[18]周彤.复方紫草凝胶剂的制备与研究.天津:天津工业大学,2017
[19]钱徐萍,陈悦桐,杨慎浩,等.中药配伍辅酶Q10凝胶剂的制备工艺优化研究.广州化工,2018,46(18):79-84
[20]张金山,程圆圆,刘健.膝骨关节炎患者血清IL-1β、TGF-β变化及相关性研究.江西中医学院学报,2011,23(6):29-32
[21]Chaganti R K,Purdue E,Sculco T P,et al.Elevation of serum tumor necrosis factorαin patients with periprosthetic osteolysis:Aase-gontrol study.Clinical Orthopaedics and Related Research,2014,472(2):584-589
[22]Losina E,Burbine S A,Suter L G.Pharmacologic regimens for knee osteoarthritis prevention:Can they be cost-effective?.Osteoarthritis Cartilage,2014,22(3):415-430
[23]李锋,聂喜增,马湘毅.骨性关节炎患者血清和关节液中自由基水平及意义.河北医药,2007,29(1):36-37
[24]史广强,金日龙,佟志慧,等.骨性关节炎人群血清基质金属蛋白酶-13水平临床观察.中国现代医生,2010,48(1):113-115
[25]Lim H,Kim H P.Matrix metalloproteinase-13 expression in IL-1β-treated hondrocytes by activation of the p38 MAPK/c-Fos/AP-1 and JAK/STAT pathways.Arch Pharm Res,2011,34(1):109-117
[2]Mc Alindon T E,Annuru R R,Sullivan M C,et al.Oarsi guidelines for the non-surgical management of knee osteoarthritis.Osteoarthritis&Cartilage,2014,22:363-388
[3]Ondrésik M,Maia F R A,Morais A D S,et al.Management of knee osteoarthritis.Current status and future trends.Biotechnology&Bioengineering,2017,114(4):717-739
[4]Murphy N J,Eyles J P,Hunter D J.Hip Osteoarthritis:Etiopathogenesis and implications for management.Advances in Therapy,2016,33(11):1921-1946
[5]王庆甫,祁印泽,陈兆军,等.超声促透透射通络止痛中药外治膝骨性关节炎的临床观察.北京中医药大学学报,2010,33(4):283-285
[6]王庆甫,马玉峰,殷岳衫,等.低频超声促透中药对兔膝关节炎细胞因子的影响.北京中医药大学学报,2013,36(2):108-112
[7]杨黎黎,王欢,张栋,等.通络止痛方对人膝骨关节炎滑膜炎性细胞TLR4、IL-1β及NO的影响研究.中华中医药杂志,2017,32(9):4124-4226
[8]杨黎黎,王欢,张栋,等.通络止痛方对人膝骨关节炎滑膜炎症细胞上清IL-1β和TNF-α的影响.辽宁中医药杂志,2017,44(5):1040-1042
[9]Zhou X,Zhang L,Guo X,et al.A Macaca fascicularis knee osteoarthritis model developed by modified Hulth combined with Joint Scratches.Medical Science Monitor International Medical Journal of Experimental&Clinical Research,2018,24:3393-3404
[10]贺石林,王健,王净净.中医科研设计与统计学.长沙:湖南科学技术出版社,2006:48-49
[11]Nirmal P,Koppikar S,Bhondave P,et al.Influence of six medicinal herbs on collagenase-induced osteoarthritis in rats.Am J Chin Med,2013,41(6):1407-1425
[12]邓莉,王海波.膝骨性关节炎动物模型的研究进展.广东医学,2018,39(12):1895-1897
[13]张君涛,吴超超,张栋林,等.膝关节骨性关节炎动物模型研究进展.重庆医学,2018,47(15):2061-2063,2067
[14]章海凤,陈树涛,冒姣娜,等.热敏灸对膝骨性关节炎兔模型IL-1β、TNF-α、MMP-13的影响.中华中医药杂志,2018,33(9):3913-3917
[15]Qinglu Luo,Xi Qin,Yaxian Qiu,et al.The change of synovial fluid proteome in rabbit surgery-induced model of knee osteoarthritis.Am J Transl Res,2018,10(7):2087-2101
[16]肖春苟,邹锦慧,李力强,等.以改良Hulth法建立兔膝骨性关节炎关节软骨的病理变化.中国中医骨伤科杂志,2014,2(12):1-3
[17]崔和平,郭兴凤.多不饱和脂肪酸对人体神经系统保健作用研究进展.河南工业大学学报(自然科学版),2012,33(3):97-102
[18]周彤.复方紫草凝胶剂的制备与研究.天津:天津工业大学,2017
[19]钱徐萍,陈悦桐,杨慎浩,等.中药配伍辅酶Q10凝胶剂的制备工艺优化研究.广州化工,2018,46(18):79-84
[20]张金山,程圆圆,刘健.膝骨关节炎患者血清IL-1β、TGF-β变化及相关性研究.江西中医学院学报,2011,23(6):29-32
[21]Chaganti R K,Purdue E,Sculco T P,et al.Elevation of serum tumor necrosis factorαin patients with periprosthetic osteolysis:Aase-gontrol study.Clinical Orthopaedics and Related Research,2014,472(2):584-589
[22]Losina E,Burbine S A,Suter L G.Pharmacologic regimens for knee osteoarthritis prevention:Can they be cost-effective?.Osteoarthritis Cartilage,2014,22(3):415-430
[23]李锋,聂喜增,马湘毅.骨性关节炎患者血清和关节液中自由基水平及意义.河北医药,2007,29(1):36-37
[24]史广强,金日龙,佟志慧,等.骨性关节炎人群血清基质金属蛋白酶-13水平临床观察.中国现代医生,2010,48(1):113-115
[25]Lim H,Kim H P.Matrix metalloproteinase-13 expression in IL-1β-treated hondrocytes by activation of the p38 MAPK/c-Fos/AP-1 and JAK/STAT pathways.Arch Pharm Res,2011,34(1):109-117