丹参水提液对碘乙酸钠所致大鼠骨关节软骨退变的预防作用
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摘要
目的探讨丹参水提液对碘乙酸钠所致大鼠骨关节软骨退变的预防作用。方法♀SD大鼠27只随机分为假手术组、碘乙酸钠组、丹参水提液组。假手术组左膝关节腔内注射生理盐水,碘乙酸钠和丹参水提液组注射碘乙酸钠(60 g·L~(-1))。注射碘乙酸钠后d 2开始灌胃给药,假手术组和碘乙酸钠组给予生理盐水(5 mL·kg~(-1)),丹参水提液组给予丹参水提液(5 mL·kg~(-1)),每天给药1次,连续6周。实验结束后处死大鼠,取血清检测MMP-13、IL-6、TNF-α;取股骨进行番红O-固绿染色;取胫骨进行Micro-CT检测。结果与假手术组相比,碘乙酸钠组血清MMP-13和股骨内侧关节软骨病理学评分升高(P<0.01),胫骨内侧关节的软骨厚度和软骨体积减低(P<0.01)。与碘乙酸钠组相比,丹参水提液组血清MMP-13和股骨内侧关节软骨病理学评分降低(P<0.05,P<0.01),胫骨内侧关节的软骨厚度和软骨体积增加(P<0.01,P<0.05)。结论丹参水提液对碘乙酸钠所致大鼠骨关节软骨退变有预防作用。
Aim To explore the preventive effect of water extract of Danshen(DWE) on degeneration of osteoarticular cartilage induced by monosodium iodocaetate(MIA).Methods Twenty-seven female SD ratswere randomly divided into sham group, MIA group and DWE group.Rats in sham group were injected a single intra-articular of saline in left knee,while rats in MIA and DWE group were injected a single intra-articular of MIA( 60 g·L-1). The rats were administered on the second day after the injection of MIA. The rats in sham group and MIA group were given 5 mL ·kg-1 of saline,and the rats in DWE group received 5 mL ·kg~(-1) of DWE every day for six weeks. At the end of the experiment,the serum was tested for MMP-13,IL-6 and TNF-α by enzyme-linked immunosorbent assay( ELISA). The left knee femur of rats was taken for safranine O-fixed green staining and tibia was collected for Micro-CT assay. Results Compared with sham group,the serum level of MMP-13 and the pathological score of the medial femoral articular cartilage in MIA group significantly increased( P < 0. 01),while cartilage thickness and cartilage volume of the medial tibial articular cartilage markedly decreased( P < 0. 01).Compared with MIA group,the serum level of MMP-13 and the pathological score of the medial femoral articular cartilage in DWE group significantly decreased( P< 0. 05,P < 0. 01),while cartilage thickness and cartilage volume of the medial tibial articular cartilage increased( P < 0. 01,P < 0. 05). Conclusion DWE has a protective effect on degeneration of osteoarticular cartilage induced by monosodium iodocaetate.
Aim To explore the preventive effect of water extract of Danshen(DWE) on degeneration of osteoarticular cartilage induced by monosodium iodocaetate(MIA).Methods Twenty-seven female SD ratswere randomly divided into sham group, MIA group and DWE group.Rats in sham group were injected a single intra-articular of saline in left knee,while rats in MIA and DWE group were injected a single intra-articular of MIA( 60 g·L-1). The rats were administered on the second day after the injection of MIA. The rats in sham group and MIA group were given 5 mL ·kg-1 of saline,and the rats in DWE group received 5 mL ·kg~(-1) of DWE every day for six weeks. At the end of the experiment,the serum was tested for MMP-13,IL-6 and TNF-α by enzyme-linked immunosorbent assay( ELISA). The left knee femur of rats was taken for safranine O-fixed green staining and tibia was collected for Micro-CT assay. Results Compared with sham group,the serum level of MMP-13 and the pathological score of the medial femoral articular cartilage in MIA group significantly increased( P < 0. 01),while cartilage thickness and cartilage volume of the medial tibial articular cartilage markedly decreased( P < 0. 01).Compared with MIA group,the serum level of MMP-13 and the pathological score of the medial femoral articular cartilage in DWE group significantly decreased( P< 0. 05,P < 0. 01),while cartilage thickness and cartilage volume of the medial tibial articular cartilage increased( P < 0. 01,P < 0. 05). Conclusion DWE has a protective effect on degeneration of osteoarticular cartilage induced by monosodium iodocaetate.
引文
[1] 钱春美,沈艳婷,袁秀荣,等.骨性关节炎研究进展[J].吉林中医药, 2016, 36(5): 538-40.[1] Qian C M, Shen Y T, Yuan X R, et al. Research progress of osteoarthritis[J]. Jilin J Tradit Chin Med, 2016, 36(5): 538-40.
[2] 杨毅峰, 黄健.骨性关节炎的流行病学研究进展[J].医学综述, 2017, 23(3): 497-501.[2] Yang Y F, Huang J. Research progress in epidemiological of osteoarthritis[J]. Med Recapitulate, 2017, 23(3): 497-501.
[3] 曹慧敏,吴瑾,贾连群,等.丹参酮ⅡA对心血管系统药理作用的研究进展[J].世界中医药, 2017, 12(7): 1718-22.[3] Cao H M, Wu J, Jia L Q, et al. Progress in study of tanshinone ⅡA′s pharmacological effects on cardiovascular system[J]. World Chin Med, 2017, 12(7): 1718-22.
[4] 庄园.丹参酮对膝骨性关节炎模型细胞活性及氧自由基代谢的影响[D].暨南大学,2007.[4] Zhang Y. The effect of tanshinone(TAN) on the activity and oxygen raicals of osteoarthritis model chondrocytes[D]. Jinan University, 2007.
[5] 徐西林, 张晓峰, 吴兴杰,等.丹参注射液对体外培养兔膝关节软骨细胞MMP-9、TIMP-1表达的影响[J].中医药学报, 2017, 45(4): 21-4.[5] Xu X L, Zhang X F, Wu X J, et al. Effect of salvia miltiorrhiza injection on expression of MMP-9 and TIMP-1 in rabbit knee joint chondrocytes cultured in vitro[J]. Acta Chin Med Pharmacol, 2017, 45(4): 21-4.
[6] 段洪超,王庆超,张杰.丹参注射液对兔软骨细胞增殖与凋亡的影响[J].西部中医药, 2018, 31(7): 22-6.[6] Duan H C, Wang Q C, Zhang J. Effects of DanShen injection on apoptosis and proliferation of rabbit chondrocytes[J]. Western J Tradit Chin Med, 2018, 31(7): 22-6.
[7] Guzman R E, Evans M G, Bove S, et al. Mono-iodoacetate-induced histologic changes in subchondral bone and articular cartilage of rat femorotibial joints: an animal model of osteoarthritis[J]. Toxicol Pathol, 2003, 31(6): 619-24.
[8] Bagi C M, Berryman E, Zakur D E, et al. Effect of antiresorptive and anabolic bone therapy on development of osteoarthritis in a posttraumatic rat model of OA[J].Arthritis Res Ther, 2015, 17: 315.
[9] Zhang Z, Wei X, Gao J, et al. Intra-articular injection of cross-linked hyaluronic acid-dexamethasone hydrogel attenuates osteoarthritis: an experimental study in a rat model of osteoarthritis[J].Int J Mol Sci, 2016, 17(4): 411.
[10] Pearson R G, Kurien T, Shu K S, et al. Histopathology grading systems for characterisation of human knee osteoarthritis-reproducibility, variability, reliability, correlation, and validity[J]. Osteoarthritis Cartilage, 2011, 19(3): 324-31
[11] 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[J].重庆医学, 2018, 47(15): 2061-3.[11] Zhang J T, Wu C C, Zhang D L, et al. Advances in animal models of knee osteoarthritis [J]. Chongqing Med, 2018, 47(15): 2061-3.
[12] Fitzpatrick L R, Green C, Maines L W, et al. Experimental osteoarthritis in rats is attenuated by ABC294640, a selective inhibitor of sphingosine kinase-2[J]. Pharmacology, 2011, 87(3-4): 135-43.
[13] Guingamp C, Gegout-Pottie P, Philippe L, et al. Mono-iodoacetate-induced experimental osteoarthritis: a dose-response study of loss of mobility, morphology, and biochemistry[J]. Arthritis Rheum, 1997, 40(9): 1670-9.
[14] 姜丽平,李龙婕,宫德正,等. 碘乙酸钠诱导原代大鼠软骨细胞凋亡[J]. 中国组织工程研究, 2013, 17(2): 247-53.[14] Jiang L P, Li L J, Gong D Z, et al. Monosodium iodoacetate induces apoptosis of primary rat chondrocytes[J]. Chin J Tissue Engineering Res, 2013, 17(2): 247-53.
[15] 李淑惠, 梁燕龙, 杨亚军,等. 丹参水溶性成分影响骨代谢的机制研究进展[J]. 中国药理学通报, 2016, 32(7): 902-5.[15] Li S H, Liang Y L, Yang Y J, et al. Research progress of regulatory effects of water-soluble compounds isolated form Salviamiltiorhiza on bone metabolism[J]. Chin Pharmacol Bull, 2016, 32(7): 902-5.
[2] 杨毅峰, 黄健.骨性关节炎的流行病学研究进展[J].医学综述, 2017, 23(3): 497-501.[2] Yang Y F, Huang J. Research progress in epidemiological of osteoarthritis[J]. Med Recapitulate, 2017, 23(3): 497-501.
[3] 曹慧敏,吴瑾,贾连群,等.丹参酮ⅡA对心血管系统药理作用的研究进展[J].世界中医药, 2017, 12(7): 1718-22.[3] Cao H M, Wu J, Jia L Q, et al. Progress in study of tanshinone ⅡA′s pharmacological effects on cardiovascular system[J]. World Chin Med, 2017, 12(7): 1718-22.
[4] 庄园.丹参酮对膝骨性关节炎模型细胞活性及氧自由基代谢的影响[D].暨南大学,2007.[4] Zhang Y. The effect of tanshinone(TAN) on the activity and oxygen raicals of osteoarthritis model chondrocytes[D]. Jinan University, 2007.
[5] 徐西林, 张晓峰, 吴兴杰,等.丹参注射液对体外培养兔膝关节软骨细胞MMP-9、TIMP-1表达的影响[J].中医药学报, 2017, 45(4): 21-4.[5] Xu X L, Zhang X F, Wu X J, et al. Effect of salvia miltiorrhiza injection on expression of MMP-9 and TIMP-1 in rabbit knee joint chondrocytes cultured in vitro[J]. Acta Chin Med Pharmacol, 2017, 45(4): 21-4.
[6] 段洪超,王庆超,张杰.丹参注射液对兔软骨细胞增殖与凋亡的影响[J].西部中医药, 2018, 31(7): 22-6.[6] Duan H C, Wang Q C, Zhang J. Effects of DanShen injection on apoptosis and proliferation of rabbit chondrocytes[J]. Western J Tradit Chin Med, 2018, 31(7): 22-6.
[7] Guzman R E, Evans M G, Bove S, et al. Mono-iodoacetate-induced histologic changes in subchondral bone and articular cartilage of rat femorotibial joints: an animal model of osteoarthritis[J]. Toxicol Pathol, 2003, 31(6): 619-24.
[8] Bagi C M, Berryman E, Zakur D E, et al. Effect of antiresorptive and anabolic bone therapy on development of osteoarthritis in a posttraumatic rat model of OA[J].Arthritis Res Ther, 2015, 17: 315.
[9] Zhang Z, Wei X, Gao J, et al. Intra-articular injection of cross-linked hyaluronic acid-dexamethasone hydrogel attenuates osteoarthritis: an experimental study in a rat model of osteoarthritis[J].Int J Mol Sci, 2016, 17(4): 411.
[10] Pearson R G, Kurien T, Shu K S, et al. Histopathology grading systems for characterisation of human knee osteoarthritis-reproducibility, variability, reliability, correlation, and validity[J]. Osteoarthritis Cartilage, 2011, 19(3): 324-31
[11] 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[J].重庆医学, 2018, 47(15): 2061-3.[11] Zhang J T, Wu C C, Zhang D L, et al. Advances in animal models of knee osteoarthritis [J]. Chongqing Med, 2018, 47(15): 2061-3.
[12] Fitzpatrick L R, Green C, Maines L W, et al. Experimental osteoarthritis in rats is attenuated by ABC294640, a selective inhibitor of sphingosine kinase-2[J]. Pharmacology, 2011, 87(3-4): 135-43.
[13] Guingamp C, Gegout-Pottie P, Philippe L, et al. Mono-iodoacetate-induced experimental osteoarthritis: a dose-response study of loss of mobility, morphology, and biochemistry[J]. Arthritis Rheum, 1997, 40(9): 1670-9.
[14] 姜丽平,李龙婕,宫德正,等. 碘乙酸钠诱导原代大鼠软骨细胞凋亡[J]. 中国组织工程研究, 2013, 17(2): 247-53.[14] Jiang L P, Li L J, Gong D Z, et al. Monosodium iodoacetate induces apoptosis of primary rat chondrocytes[J]. Chin J Tissue Engineering Res, 2013, 17(2): 247-53.
[15] 李淑惠, 梁燕龙, 杨亚军,等. 丹参水溶性成分影响骨代谢的机制研究进展[J]. 中国药理学通报, 2016, 32(7): 902-5.[15] Li S H, Liang Y L, Yang Y J, et al. Research progress of regulatory effects of water-soluble compounds isolated form Salviamiltiorhiza on bone metabolism[J]. Chin Pharmacol Bull, 2016, 32(7): 902-5.