中药复方及其拆方制剂对寒冷刺激结合石膏固定骨关节炎模型大鼠血清和膝关节滑膜组织中相关炎性因子的影响

摘要

目的 探讨中药复方及其拆方制剂对寒冷刺激结合石膏同定骨关节炎模型大鼠血清和膝关节滑膜组织中相关炎性因子的影响.方法 选取无特定病原体级Sprague Dawley大鼠60只,按照随机数字表法分为空白对照组、模型对照组、西药组、中药复方组、中药拆方1组、中药拆方2组,各10只.除空白对照组外,其余5组采用寒冷刺激结合石膏固定的方法制作骨关节炎大鼠模型.空白对照组、模型对照组大鼠给予与用药组同容积0.9％氯化钠注射液灌胃,中药复方组、中药拆方1组及中药拆方2组均给予中药水煎剂灌胃,西药组给予塞来昔布灌胃,均为1次/d,连续给药8周,最后一次给药后,测定大鼠血清和滑膜组织中炎性细胞因子[白细胞介素1β(IL-1 β)、肿瘤坏死因子α(TNF-α)、前列腺素E2(PGE2)、环氧合酶2(COX-2)、γ干扰素]及关节软骨中聚蛋白多糖酶4(ADAMTs-4)、ADAMTs-5蛋白表达.观察比较各组大鼠膝关节滑膜组织病理学结构.结果 模型对照组、中药复方组、中药拆方1组、中药拆方2组、西药组大鼠血清及滑膜组织中IL-1β、TNF-α、PGE2、COX-2、γ干扰素水平高于空白对照组,差异均有统计学意义[血清:(19.6±2.7)、(6.5±2.4)、(20.4±2.4)、(10.3±4.5)、(15.9±2.6)ng/L比(5.9±2.2)ng/L,(72.6±7.6)、(21.8±5.9)、(78.9±1 1.4)、(43.2±2.2)、(32.9±6.6) ng/L比(5.2±2.4) ng/L,(99±11)、(36±7)、(32±7)、(70±8)、(55±8) ng/L比(30±5) ng/L,(100±13)、(24±5)、(64±8)、(63±8)、(24±7)U/L比(11 ±4) U/L,(322 ±24)、(112±18)、(206±14)、(213±8)、(324±22) ng/L比(108±16) ng/L;滑膜组织:(16.9±4.8)、(4.9±2.6)、(9.3±2.8)、(17.2±4.6)、(9.6±3.5) ng/L比(3.2±1.3) ng/L,(135.3 ±21.3)、(33.5±11.0)、(52.8±12.1)、(139.7±25.6)、(39.5±16.7) ng/L比(17.5±4.4) ng/L,(211 ±21)、(58±12)、(111 ±17)、(220±25)、(219±22) ng/L比(49±12) ng/L,(80±11)、(27±10)、(51±15)、(85±22)、(13 ±6) U/L比(11 ±3)U/L,(413±60)、(169±46)、(427 ±43)、(332±71)、(137±40) ng/L比(145 ±45) ng/L](均P＜0.05).中药复方组大鼠血清及滑膜组织中上述炎性因子水平低,西药组血清中IL-1β、TNF-d、PGE2、COX-2及滑膜组织中IL-1β、TNF-αα、COX-2、γ干扰素水平低于模型对照组,差异均有统计学意义(均P ＜0.05).西药组血清中IL-1β、TNF-α、PGE2、γ十扰素及滑膜组织中IL-1β、TNF-α、PGE2水平高于中药复方组,差异均有统计学意义(均P＜0.05).各组大鼠关节软骨ADAMTs-4、ADAMTs-5蛋白均有明显表达,模型对照组大鼠表达最为明显,中药复方组表达最低.中药复方组大鼠软骨表层较光滑,细胞排列较规则,无明显裂隙,潮线完整.结论 中药复方能够明显降低骨关节炎大鼠血清及滑膜组织中炎性因子,阻止局部关节软骨ADAMTs合成,减轻骨关节炎软骨损伤,具有较好疗效.%Objective To explore effects of compound Chinese medicine and its split preparation on inflammatory factors in serum and knee joint synovial membrane in osteoarthritis rat model.Methods Totally 60 SD rats were randomly divided into blank control group,model control group,western medicine group (W group),compound Chinese medicine group (C group),Chinese split prescription group 1 (D1 group) and Chinese split prescription group 2(D2 group),with 10 rats in each group.Rats in model control group,W group,C group,D1 group and D2 group were made osteoarthritis model induced by cold stimulation and plaster fixation.All rates were treated with intragastrical administration,with 0.9％ sodium chloride injection for blank control group and model control group,Chinese medicine decoction for C,D1,D2 groups andcelecoxib for W group,once daily for 8 weeks.Expressions of inflammatory cytokines [interleukin-1β (IL-1β),tumor necrosis factor-α (TNF-α),prostaglandin E2 (PGE2),cyclooxygenase-2 (COX-2),interferon-γ(INF-γ)] in serum and synovial membrane,expressions of ADAMTs-4(a disintegrin and metalloprotease with thrombospond in type 1 motifs) and ADAMTs-5 in articular cartilage were detected.Pathological structure of knee joint synovial membrane tissue was observed.Results Levels of IL-1β,TNF-α,PGE2,COX-2,IFN-γ in serum and synovial membrane in model control group,W group,C group,D1 group and D2 group were all significantly higher than those in blank control group[serum:(19.6 ±2.7),(6.5 ±2.4) (20.4 ±2.4),(10.3 ±4.5),(15.9 ±2.6)ng/L vs (5.9 ± 2.2)ng/L;(72.6±7.6),(21.8 ±5.9),(78.9 ± 11.4),(43.2 ±2.2),(32.9 ±6.6)ng/L vs (5.2 ± 2.4)ng/L;(99 ± 11),(36±7),(32 ±7),(70±8),(55 ±8)ng/L vs (30 ±5)ng/L;(100 ± 13),(24 ±5),(64 ±8),(63 ±8),(24 ±7)U/L vs (11 ±4)U/L;(322 ±24),(112 ± 18),(206 ± 14),(213 ±8),(324 ± 22) ng/L vs (108 ± 16) ng/L;synovial membrane:(16.9 ± 4.8),(4.9 ± 2.6),(9.3 ± 2.8),(17.2 ± 4.6),(9.6±3.5)ng/L vs (3.2±1.3)ng/L;(135.3 ±21.3),(33.5±11.0),(52.8±12.1),(139.7 ±25.6),(39.5±16.7)ng/Lvs (17.5±4.4)ng/L;(211 ±21),(58±12),(111±17),(220±25),(219±22)ng/L vs (49±12)ng/L;(80± 11),(27 ± 10),(51 ± 15),(85 ±22),(13 ±6) U/L vs (11 ± 3) U/L;(413 ±60),(169 ±46),(427 ±43),(332 ±71),(137 ± 40) ng/L vs (145 ±45) ng/L] (all P ＜0.05).Levels of IL-1β,TNF-α,PGE2,COX-2,IFN-γ in serum and synovial membrane in C group were all significantly lower than those in model control group;levels of IL-1 β,TNF-α,PGE2,COX-2 in serum and IL-1β,TNF-α,COX-2,IFN-γ in synovial membrane in W group were all significantly lower than those in model control group (all P ＜ 0.05).ADAMTs-4 and ADAMTs-5 were detected in all groups.In C group,histopathological results showed that cells in knee joint synovial membrane tissue were regularly arranged,the cartilage surface layer was smooth,with no obvious fracture and complete tidal line.Conclusion Compound Chinese medicine significantly reduces inflammatory factors in serum and synovial membrane in osteoarthritis rats,it inhibits the synthesis of ADAMTs in local articular cartilage and alleviates osteoarthritis damage.