膝三脏汤不同工艺提取物对KOA模型大鼠血清炎症因子及软骨端粒酶活性的影响
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摘要
【目的】观察膝三脏汤石油醚、氯仿、乙酸乙酯、正丁醇提取物对膝骨关节炎(KOA)大鼠模型血清炎症因子含量及软骨端粒酶活性的影响,比较不同工艺提取物的生物活性和潜力。【方法】将70只大鼠随机分为空白组、模型组、美洛昔康组、膝三脏汤—石油醚组、膝三脏汤—氯仿组、膝三脏汤—乙酸乙酯组、膝三脏汤—正丁醇组,每组10只。除空白组外,将其余各组大鼠按改良Hulth造模法复制KOA模型。造模结束后,各组对应给药。给药6周后,测定大鼠血清中白细胞介素(IL)-1、IL-6、肿瘤坏死因子(TNF)-α含量,软骨含水率和端粒酶活性。【结果】与模型组比较,各膝三脏汤提取物组大鼠血清炎症因子IL-1、IL-6、TNF-α含量、软骨含水率、端粒酶活性明显降低。其中:乙酸乙酯组大鼠血清IL-1、TNF-α含量,软骨端粒酶活性均低于氯仿组、正丁醇组(P <0.05),与石油醚组、美洛昔康组差异无统计学意义(P> 0.05);乙酸乙酯组大鼠血清IL-6含量、关节软骨含水率均低于石油醚组、正丁醇组(P <0.05),与氯仿组、美洛昔康组差异无统计学意义(P> 0.05)。【结论】膝三脏汤石油醚、氯仿、乙酸乙酯、正丁醇提取物均可有效改善KOA大鼠血清炎症反应、软骨老化情况,但以乙酸乙酯法的改善作用较为明显。
Objective To observe the effects of the extracts from Knee Triple-organ Decoction(KTD)obtained by petroleum ether,chloroform,ethyl acetate or n-butanol on the contents of serum inflammatory cytokines and the activity of cartilaginous telomerase in rat model of knee osteoarthritis(KOA), so as to compare the biological activities and potential between different KTD extracts. Methods Seventy rats were randomly divided into blank group, model group, meloxicam group, KTD-petroleum ether group, KTD-chloroform group, KTD-ethyl acetate group,KTD-n-butyl alcohol group,10 rats in each group. Except for the blank group,the rats in other groups were induced into KOA model by modified Hulth method. After modeling, all of the groups were given corresponding medicines respectively. After 6-week medication,the contents of serum IL-6,IL-1,TNF-α,water content in articular cartilage and the activity of cartilaginous telomerase were determined. Results Compared with the model group,the contents of serum inflammatory cytokines of IL-6,IL-1 and TNF-α,the water content in cartilage and the activity of cartilaginous telomerase were decreased in various KTD extracts groups. The serum contents of IL-1 and TNF-α,and the activity of cartilaginous telomerase in the ethyl acetate group were lower than those in the chloroform group and n-butyl alcohol group(P < 0.05),but were not statistically different from those in the petroleum ether group and meloxicam group(P > 0.05). The serum content of IL-6 and the water content in articular cartilage in the ethyl acetate group were lower than those in the petroleum ether group and n-butyl alcohol group(P < 0.05),but were not statistically different from those in the chloroform group and meloxicam group(P >0.05). Conclusion KTD extracts obtained by petroleum ether, chloroform, ethyl acetate or n-butanol have certain effect on the inflammatory reaction and cartilage aging,and the effect of ethyl acetate method is stronger.
Objective To observe the effects of the extracts from Knee Triple-organ Decoction(KTD)obtained by petroleum ether,chloroform,ethyl acetate or n-butanol on the contents of serum inflammatory cytokines and the activity of cartilaginous telomerase in rat model of knee osteoarthritis(KOA), so as to compare the biological activities and potential between different KTD extracts. Methods Seventy rats were randomly divided into blank group, model group, meloxicam group, KTD-petroleum ether group, KTD-chloroform group, KTD-ethyl acetate group,KTD-n-butyl alcohol group,10 rats in each group. Except for the blank group,the rats in other groups were induced into KOA model by modified Hulth method. After modeling, all of the groups were given corresponding medicines respectively. After 6-week medication,the contents of serum IL-6,IL-1,TNF-α,water content in articular cartilage and the activity of cartilaginous telomerase were determined. Results Compared with the model group,the contents of serum inflammatory cytokines of IL-6,IL-1 and TNF-α,the water content in cartilage and the activity of cartilaginous telomerase were decreased in various KTD extracts groups. The serum contents of IL-1 and TNF-α,and the activity of cartilaginous telomerase in the ethyl acetate group were lower than those in the chloroform group and n-butyl alcohol group(P < 0.05),but were not statistically different from those in the petroleum ether group and meloxicam group(P > 0.05). The serum content of IL-6 and the water content in articular cartilage in the ethyl acetate group were lower than those in the petroleum ether group and n-butyl alcohol group(P < 0.05),but were not statistically different from those in the chloroform group and meloxicam group(P >0.05). Conclusion KTD extracts obtained by petroleum ether, chloroform, ethyl acetate or n-butanol have certain effect on the inflammatory reaction and cartilage aging,and the effect of ethyl acetate method is stronger.
引文
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[14] Sakao K, Takahashi K A, Mazda O, et a1. Enhanced expression of interleukin-6,matrix metalloproteinase-13,and receptor activator of NF-kappaB ligand in cells derived from osteoarthritic subchondral bone[J]. J Orthop Sci, 2008, 13(3):202.
[15] Westacott C I,Sharif M. Cytokines in osteoarthritis:mediators or markers of joint destruction?[J]. Semin Arthritis Rheum,1996,25(4):254.
[16] Solchaga L A,Dennis J E,Goldberg V M,et al. Hyaluronic acid-based polymers as cell carriers for tissue-engineered repair of bone and cartilage[J]. J Orthop Res,1999,17(2):205.
[17]谢建明,王好问,陆才生. TNF-α上调单核巨噬细胞MMP-9的活性与类风湿关节炎关节破坏的关系[J].中国病理生理杂志,2009,25(6):1181.
[18]邓廉夫. IL-1、TNF和IL-6与骨关节炎[J].国际骨科学杂志,1996(2):102.
[19]陈连旭,余家阔,于长隆,等.大鼠、兔和人关节软骨组织形态学的比较[J].中国组织工程研究与临床康复,2007,11(41):8230.
[20] Blanco F J,Guitian R,Vázquez-Martul E,et al. Osteoarthritis chondrocytes die by apoptosis. Apossible pathway for osteoarthritis pathology[J]. Arthritis Rheum, 1998, 41(2):284.
[2]吴栓,邢振龙,黄辉文,等.基于“三脏一体”辨证理论治疗膝骨性关节炎疗效分析[J].中国中医骨伤科杂志,2014,22(3):22.
[3]陈水昌,邢振龙,丘青中.膝三脏汤在膝关节骨性关节炎中的应用[J].河北中医,2016,38(3):373.
[4]邢振龙,丘青中,吴栓.“三脏一体”整体观在膝骨性关节炎治疗中的应用[J].中国中医骨伤科杂志,2014,22(10):38.
[5]丘青中,邢振龙,戚子荣.膝三脏汤对膝骨关节炎大鼠炎症因子及关节软骨含水率影响实验研究[J].辽宁中医药大学学报,2017,19(12):25.
[6]刘献祥,李西海,周江涛.改良Hulth造模法复制膝骨性关节炎的实验研究[J].中国中西医结合杂志,2005,25(12):1104.
[7] Sharpless N E, DePinho R A. Telomerase, stem cells, senescence and cancer[J]. J Clin Invest,2004,113:160.
[8] Al-Hajj M, Wicha M S, Benito H A, et al. Prospective identification of tumorigenic breast cancer cells[J]. Proc Natl Acad Sci USA,2003,100:3983.
[9]卫立辛,郭亚军,闫振林,等.检测人端粒酶活性的端粒酶TRAP-ELISA法的建立[J].中华肿瘤杂志,1998,20(6):465.
[10] Calich A L, Domiciano D S, Fuller R. Osteoarthritis:can anti-cytokine therapy play a role in treatment?[J]. Clin Rheumatol,2010,29(5):451.
[11] Raymond L, Eck S, Hays E, et al. RelA is required for IL-1beta stimulation of matrix metalloproteinase-1 expression in chondrocytes[J]. Osteoarthritis Cartilage,2007,15(4):431.
[12] Legendre F, BaugéC, Roche R, et al. Chondroitin sulfate modulation of matrix and inflammatory gene expression in IL-1beta-stimulated chondrocytes--study in hypoxic alginate bead cultures[J]. Osteoarthritis Cartilage,2008,16(1):105.
[13] Bjornsson G L,Thorsteinsson L,Gudmundsson K O,et al.Inflammatory cytokines in relation to adrenal response following total hip replacement[J]. Scand J Immunol,2007,65(1):99.
[14] Sakao K, Takahashi K A, Mazda O, et a1. Enhanced expression of interleukin-6,matrix metalloproteinase-13,and receptor activator of NF-kappaB ligand in cells derived from osteoarthritic subchondral bone[J]. J Orthop Sci, 2008, 13(3):202.
[15] Westacott C I,Sharif M. Cytokines in osteoarthritis:mediators or markers of joint destruction?[J]. Semin Arthritis Rheum,1996,25(4):254.
[16] Solchaga L A,Dennis J E,Goldberg V M,et al. Hyaluronic acid-based polymers as cell carriers for tissue-engineered repair of bone and cartilage[J]. J Orthop Res,1999,17(2):205.
[17]谢建明,王好问,陆才生. TNF-α上调单核巨噬细胞MMP-9的活性与类风湿关节炎关节破坏的关系[J].中国病理生理杂志,2009,25(6):1181.
[18]邓廉夫. IL-1、TNF和IL-6与骨关节炎[J].国际骨科学杂志,1996(2):102.
[19]陈连旭,余家阔,于长隆,等.大鼠、兔和人关节软骨组织形态学的比较[J].中国组织工程研究与临床康复,2007,11(41):8230.
[20] Blanco F J,Guitian R,Vázquez-Martul E,et al. Osteoarthritis chondrocytes die by apoptosis. Apossible pathway for osteoarthritis pathology[J]. Arthritis Rheum, 1998, 41(2):284.