丹参对缺血肢体关节软骨及滑膜损伤的影响

目的　通过动物实验观察丹参关节内注射对关节缺血再灌注损伤后退行性关节炎的防治作用。方法　 6 3只新西兰大白兔随机分成对照、缺血 2h ,缺血 5h用丹参和不用丹参 5个组 ,分别于缺血、再灌注 1d和 1、 4、 8、 12、 16周取股骨外髁软骨及滑膜行大体和光镜观察 ,以及再灌注后 8周和 16周软骨厚度测量。结果　实验组关节软骨、滑膜早期有缺血再灌注损伤 ,并逐渐演变成早期退行性关节炎改变 ,而这些病变用丹参组明显轻于未用丹参组。结论　丹参能有效地减轻关节软骨及滑膜缺血再灌注损伤及退行性改变。     

丹参与缺血肢体关节软骨及滑膜损伤的影响

目的 通过动物实验观察丹参关节内注射对关节缺血再灌注损伤后退行性关节炎的防治作用。方法 ６３只新西兰大白兔随机分成对照，缺血２ｈ，缺血５ｈ用丹参和不同丹参５个组，分别于缺血，再灌注１ｄ和１、４、８、１２、１６周取股骨外髁软骨及滑膜行大体和光镜观察，以及再灌注后８周和１６周软骨厚度测量。结果 实验组关节软骨，滑膜早期有缺血再灌注损伤，并逐渐演变成早期退行性关节炎改变，而这些病变用对丹参组。结论 丹参     

肢体缺血再灌注后关节软骨及滑膜损伤的组织学观察

目的：通过动物实验观察肢体缺血再灌注后关节软骨和滑膜病理变化。方法：35只新西兰大白兔随机分成缺血2、5h和正常对照3个组。分别于缺血、再灌注后1d和1、4、8、12、16周取关节软骨和滑膜行大体和光镜观察,测量8和16周软骨各层厚度,并进行定量分析。结果：肢体缺血再灌注1周内关节软骨及滑膜表现为缺血再灌注损伤,4周后渐出现退行性关节炎的早期改变,缺血5h组重于缺血2h组。结论：缺血再灌注肢体关节软骨、滑膜有缺血再灌注损伤,可逐渐演变成骨性关节炎。     

肢体缺血再灌注后关节软骨的组织学观察

目的: 通过动物实验观察肢体缺血再灌注后关节软骨的病理变化, 证实缺血再灌注导致关节软骨的损伤, 并探讨关节缺血再灌注损伤与骨性关节炎形成的关系。方法: 健康中国本地兔42只, 随机分为正常对照组, 缺血4h组和缺血10h组, 分别于肢体缺血再灌注后0、1d和1、5、10周,取4只兔8条腿做标本, 行光镜观察, 及图像分析仪处理。结果: 早期(24h以内) 关节软骨的组织学改变在光镜下表现不明显。再灌注5周时损伤最为明显, 此后短期内(10周内) 软骨损伤不再加重。软骨组织形态学的定量观察,股骨外髁关节软骨全层厚度明显增加(P<0 .01), 非钙化层增加不明显, 而钙化层厚度则明显增加(P< 0. 01)。结论: (1) 证实了缺血再灌注导致关节软骨的损伤; (2) 关节缺血再灌注损伤也是导致骨性关节炎形成的重要原因。     

风湿骨痛药酒药槌外治法防治关节软骨退变的实验研究

目的 探讨风湿骨痛药酒药槌外治法防治关节软骨退变的作用机理。方法 选用日本大耳白兔18只，随机将12只通过结扎右股静脉造成膝关节骨性关节炎模型。未造模兔6只为正常组，12只造模兔随机分为造模组和治疗组。造模8周后，治疗组施以风湿骨痛药酒槌外治法进行治疗。造模11周后，观察软骨病理组织形态学、滑膜中NO浓度、软骨中PA及PAI活性和关节液中HA含量变化。结果 治疗组滑膜中NO浓度、软骨中PA及PAI活性和关节液中HA含量与正常组相比有显著性差异。造模组所测各项指标与治疗组相比有显著性差异。治疗组滑膜中NO浓度及软骨中PA含量明显低于造模组，而软骨中PAI含量及关节液中HA含量明显高于造模组。治疗组关节软骨退变不明显，类似正常组，而造模组关节软骨退行性变明显。结论 风湿骨痛药酒药槌外治法能通过改善骨内及周围组织的微循环，抑制滑膜组织中NO的过度产生，降低软骨中PA的活性，提高PAI的活性，消除导致关节软骨退变的内在因素，提高关节液中HA的含量，从而达到保护关节软骨、抗关节软骨退变的目的。     

5氟脲嘧啶关节腔注射治疗兔膝骨性关节炎的实验研究

[目的]评价5氟脲嘧啶（5-Fluorouracil，5Fu）关节腔注射治疗兔膝骨性关节炎（osteoarthritis，OA）的疗效。[方法]24只兔子制成骨关节炎模型随机分成OA组、5Fu组和对照组，OA组立即处死，5Fu组按5Fu2ms／kg关节腔注射，每周1次连续4次，对照组注射等量生理盐水，最后1次治疗后1周处死。观察3组滑膜组织的光镜、电镜改变及软骨的光镜、MMP-1免疫组化改变，比较软骨Mankin＇s评分及关节液中IL-1的浓度。[结果]5Fu组可见软骨破坏减轻，滑膜炎症明显抑制，Mankin’s评分明显改善（P〈0．01）；关节液IL-1浓度降低（P〈0．05），关节软骨中MMP-1表达减弱。[结论]5Fu关节腔内注射能抑制滑膜炎症，缓解软骨的破坏。     

大黄素对大鼠肝脏缺血再灌注损伤的预防作用

目的:探讨大黄素对肝脏缺血-再灌注损伤的保护作用及其机制.方法:将45只成年雄性SD大鼠随机分成三组:假手术对照组(A组)15只,肝缺血30 min、再灌注90min组(B组)15只,术前5 d给予大黄素灌胃60mg/(kg·d),5次 肝缺血30 min、再灌注90min(C组)15只;观察血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、透明质酸酶(HA)、肝组织丙二醛(MDA)含量的变化及肝脏组织病理学改变情况,同时测定各组大鼠胆汁流量情况.结果:C组与B组相比,MDA含量显著降低(P＜0.05),HA浓度明显降低(P＜0.01),胆汁流量显著增加(P＜0.01),同时C组的肝细胞功能明显改善,且肝脏超微结构改变也较轻.结论:大黄素对肝脏缺血-再灌注损伤具有保护作用,该作用与减轻肝脏缺血再灌注后脂质过氧化程度和肝窦内皮细胞损伤有关.     

臭氧液对兔膝骨性关节炎滑膜炎症的影响

目的评价臭氧液对兔膝骨性关节炎滑膜炎症的影响。方法新西兰兔32只,采用随机数字表法分为四组,每组8只。A组不予任何处理;B组仅造模;C组造模成功后,关节腔注射20μg/ml臭氧液2ml,每周1次,连续3周;D组不造模,仅行关节腔注射臭氧液,方法同C组。观察膝关节滑膜形态,检测滑膜组织中IL-6和TNF-α含量。结果 A组滑膜组织无充血水肿,滑膜细胞无增生现象,滑膜组织结构正常;B组滑膜组织正常结构破坏,细胞重度增生,炎性细胞浸润明显,血管增生充血,滑膜厚度较A组明显增加;C组滑膜组织炎症细胞减少,充血水肿减轻,滑膜厚度较B组明显降低;D组滑膜无炎症表现。与A组比较,D组IL-6和TNF-α含量无明显增高,B、C组IL-6和TNF-α含量明显增高(P0.05),但C组明显低于B组(P0.05)。结论关节腔注射20μg/ml臭氧液2ml可以减轻滑膜炎症,降低滑膜中IL-6和TNF-α含量,且对正常关节滑膜组织无明显损害。     

小白菊内酯不同给药途径对兔膝骨关节炎的实验研究

目的 通过测定膝骨关节炎兔模型血清和关节液中TNF-α、IL-1β的含量,观察软骨的组织学改变,探讨小白菊内酯不同给药途径治疗骨关节炎的疗效与作用机制。方法 健康纯种新西兰兔40只,随机取8只作为正常组（A组）;其余的32只采用右后肢伸直位管型石膏固定法,建立OA模型。造模后随机分为4组：模型口服对照组（B组）、模型关节腔注射对照组（C）、小白菊内酯口服组（D组）、小白菊内酯关节腔注射组（E组）,每组8只。B组和D组兔分别予生理盐水、小白菊内酯灌胃,C组和E组兔分别予生理盐水、小白菊内酯关节腔注射,连续治疗6w后,测定各组兔治疗前后血清、膝关节液TNF-α、IL-1β表达水平。所有实验动物在8w处死,应用苏木素-伊红（HE）染色观察各组动物关节软骨的病理组织改变。结果 与模型组（B组、C组）比较,D组、E组关节液及血清中的TNF-α、IL-1β浓度均明显降低(P< 0.05),且E组降低幅度明显大于D组(P<0.05)。与A组比较,B组和C组病理显示有关节软骨破坏,病理积分升高（P<0.05）;D组、E组病理改变较B组、C组显示关节软骨有所修复,病理积分降低（P<0.05）。结论 小白菊内酯可减轻动物模型的关节炎症,其关节腔给药的抗炎疗效优于口服给药,抑制血清和关节腔TNF-α、IL-1β的分泌,可能是小白菊内酯治疗骨关节炎的机制之一。     

MMP-3／TIMP-1在大鼠肢体再灌注后关节软骨损伤的作用

目的 探讨缺血再灌注后关节软骨中MMP-3／TIMP-1比例变化与软骨损伤的关系。方法 采用大鼠后肢股动脉夹闭的方法模拟缺血再灌注的动物模型，用Wistar大鼠40只，随机分成正常对照组(NG)、肢体单纯缺血组(IG)和缺血再灌注组(IR)。运用免疫组化技术，分别测定TIMP-1和MMP-3在关节软骨中不同时相的表达变化并进行半定量分析，观察关节软骨病理改变及蛋白多糖(PG)的变化。结果 缺血再灌注后，关节软骨中的MMP-3和TIMP-1表达均有增加，但MMP-3增加的幅度大于TIMP，导致MMP-3／TIMP-1比值增大，与再灌注后引起的关节软骨损伤相关。结论 MMP／TIMP的失平衡表达是导致缺血再灌注后关节软骨损伤的重要因素。     

Repair of articular cartilage injury following intra-articular chymopapain-induced matrix proteoglycan loss

The intra-articular injection of 0.02, 0.2, or 2.0 mg of chymopapain (CP) into the knee of adolescent rabbits caused the loss of more than 50% of the proteoglycans (PGs) in the cartilaginous tissues within the joint. Sequential measurements of cartilage-derived keratan sulfate epitope in serum and analyses of articular cartilage slices 2 days after the injection revealed that 0.02 mg of CP was nearly as effective as higher doses (0.2 or 2.0 mg of CP) in causing the depletion. The degradation and depletion of PGs in articular cartilage were shown to be localized to the joint and did not affect articular cartilage in the contralateral knee joint (no injection) or other cartilaginous tissues in the body. On day 9, partial replenishment of the articular cartilage PGs had occurred, irrespective of the dose used, and the articular surface within the joint remained intact. However, by day 21, articular cartilage in joints injected with 2.0 mg of CP had begun to show progressive degenerative changes, and these changes became more severe with time. In contrast, joints injected with 0.2 mg of CP continued to repair successfully by the reestablishment of a matrix that retained its integrity and appeared to remain functional for at least 6 months. These results suggest that the model may prove useful for the study of the repair processes that follow matrix injury and severe depletion of PGs from the articular cartilage matrix.     

Hyaluronic acid attenuates osteoarthritis development in the anterior cruciate ligament-transected knee: Association with excitatory amino acid release in the joint dialysate.

We previously reported increased release of the excitatory amino acid (EAA) neurotransmitters, glutamate and aspartate, during the early stage of experimental osteoarthritis (OA). Our present objective was to study the effect of intraarticular injection of hyaluronic acid (HA) on OA development, and to analyze concomitant changes in EAA levels in dialysates of anterior cruciate ligament-transected (ACLT) knee joints. OA was induced in Wistar rats by ACLT of one hindlimb; the knee of the other hindlimb was used as the sham-operated control. HA group (n = 12) were injected intraarticularly in the ACLT knee with 1 mg of HA once a week for 5 consecutive weeks, starting at 8 weeks after surgery. Saline group (n = 12) were injected as above with normal saline. The sham-operated group, underwent arthrotomy, but not ACLT, and received no treatment (n = 14). Twenty weeks after surgery, knee joint dialysates were collected by microdialysis and EAA levels assayed by high-performance liquid chromatography, and gross morphological examination and histopathological evaluation were performed on the medial femoral condyles and synovia. Rats receiving intraarticular HA injections showed a significantly lower degree of cartilage degeneration on the medial femoral condyle at both the macroscopic level and on the Mankin grading scale than rats receiving saline injections. Intraarticular HA treatment also suppressed synovitis. Moreover, glutamate and aspartate levels were significantly reduced in the HA group compared to the saline group. Intraarticular injection of HA limits articular cartilage and synovium damage and OA formation, and, in parallel, reduces EAA levels in ACLT joint dialysates. This study suggests that the underlying mechanism of the anti-inflammatory effect of HA is to inhibit glutamate and aspartate release in ACLT knee joints, which attenuates the early development of OA.     

丹参和透明质酸钠局部给药对兔骨关节炎的影响

目的探讨丹参和透明质酸钠局部给药对兔膝骨关节炎(OA)模型的影响。方法建立Hulth兔膝OA动物模型。每周经膝关节穿刺给药1次,观察丹参和透明质酸钠单用或合用对兔膝关节组织形态学、滑膜丙二醛(MDA)含量及血红细胞超氧化物歧化酶(SOD)比活性的影响。结果丹参或透明质酸钠单用和合用组均能显著降低兔膝关节大体软骨分级评分和镜下Mankin’s分级评分(P<0.01),尤以合用组作用最佳;同时丹参单用或与透明质酸钠合用组还明显增加红细胞SOD比活性(103%,158%),减少滑膜MDA生成量(66%,77%)。结论丹参和透明质酸钠局部注射对OA有治疗作用,尤以合用时显著。     

关节腔冲洗并注射透明质酸钠治疗轻度膝关节炎70例

目的探讨关节腔冲洗并注射透明质酸钠治疗轻度膝关节炎（knee osteoarthritis,KOA）的临床疗效。方法 2007年7月至2009年3月共收治70例KOA患者。随机分为两组,治疗组35例,采用关节腔冲洗配合透明质酸纳关节腔注射的二联疗法;对照组35例,单用玻璃酸钠关节腔注射。将两组治疗3个月和2年的疗效随访进行对比观察。结果全部获得随访,随访时间为0.5～2年,平均14个月。治疗后3个月随访,治疗组治愈率为82.8%,对照组为60%（P〈0.01）;治疗后末次随访,治疗组复发率为22%,对照组为37%（P〈0.01）。结论二联疗法是治疗KOA的一种简便、安全可靠、无副作用、疗效确切的方法。     

兔实验性膝骨性关节炎的组织病理学及发病机理

目的：探讨骨性关节炎发生的病理学基础。方法：40只健康雄性家兔随机分为对照组和模型组。模型组用Hulth法复制出实验性膝骨性关节炎模型。对照组不作任何处理。1月后处死家兔并切取家兔股骨内髁关节软骨作光镜及透射电镜观察,光镜所见依据Mankin's评分标准评分,测定血清中红细胞超氧化物歧化酶（SOD）活性并作统计学处理。结果：模型组家兔膝关节滑膜均存在不同程度增生、肥厚、水肿,光镜及电镜下见模型组关节软骨呈明显退行性变,Mankin's评分结果模型组得分显著高于对照组（P＜0．05）,模型组SOD活性显著低于对照组（P＜0．05）。结论：滑膜组织的炎症、骨内压的增高以及自由基的改变是导致骨性关节炎发生的病理学基础,如何阻断这些病理环节是防治骨性关节炎的关键。     

不同浓度木瓜蛋白酶建立兔膝骨关节炎模型的比较研究

目的:比较不同浓度木瓜蛋白酶建立的兔膝骨关节炎模型,为研究骨关节炎病理及诊疗方法提供数据支持。方法:将60只新西兰大耳白兔随机分为4组,分别在1、3、5d右膝关节腔注射2%(低剂量组)、5%(中剂量组)、10%(高剂量组)(w/v)木瓜蛋白酶和0.03mol/L的L-半胱氨酸混合溶液0.1ml/kg(实验组),以及等量生理盐水(对照组)。整个实验过程观察兔状态并记录体重和膝关节宽度。首次注射后第2、4、6周将兔分批处死,取股骨髁、胫骨平台及滑膜做大体及组织病理学观察,并进行评分。结果:实验组均出现关节宽度增大、关节软骨变薄、基质分解破坏、滑膜增生等炎症及退行性改变,严重程度随浓度增高而增高,也随时间延长而发展;对照组无上述改变,二者比较差异有统计学意义。结论:兔膝骨关节腔注射2%、5%、10%木瓜蛋白酶和0.03mol/LL-半胱氨酸混合溶液0.1ml/kg可制备不同严重程度的骨关节炎模型,周期短,可重复且成功率高。     

自体游离骨膜再造膝关节半月板的实验研究

目的：对于严重损伤的半月板全切除术后行自体游离骨膜重建半月板，以维持膝关节结构、生理及功能特性。方法：使用１７只成年犬，５只幼犬。外侧半月板全切除以后，胫骨内侧近端的骨膜游离并移植到外侧半月板部位。于２、４、８、１２、２４、４８周取下重建半月板及不同部位的关节面软骨进行大体、光镜及电镜观察。结果：术后２个月，重建半月板的大体形态、组织结构与正常半月板近似。外侧股骨软骨面及没有被覆盖的胫骨软骨面表现退行性改变，被移植物覆盖的软骨面退变较轻。幼犬的退变更早、更严重。结论：本研究说明了自体游离骨膜在滑液环境及受到合适的应力刺激会向纤维软骨演化，在半月板全切除后，应用游离骨膜重建半月板是可行的     

Metabolic and structural changes in newly synthesized proteoglycans induced by implantation of a polyethylene sheet in the rabbit knee joint

A sheet of polyethylene was surgically implanted in a rabbit right patello-femoral joint and changes in the structure and chemical composition of newly synthesized articular cartilage proteoglycans (PGs) were studied 1 month after surgery. The articular cartilage from implanted and sham-operated control knee joints was labeled in vitro with 3H-glycine and 35S-SO4 and then extracted with 4 M guanidinium chloride (GuHCl) solution. Labeled extracts were analyzed by dissociative CsCl gradient centrifugation and by Sepharose CL-2B column chromatography. The labeled glycosaminoglycan side chains were analyzed by Sephadex G-200 column chromatography and specific enzymatic digestions. Compared with sham-operation, the trochlear articular cartilage of operated joints incorporated more 35S-SO4 and 3H-glycine into newly synthesized PGs and proteins. It also synthesized a higher proportion of extractable, hydrodynamically large and high density 35S-PG monomers with increased proportion of molecules, able to interact with exogeneous hyaluronan (HA). The fibro-cartilagenous 'osteo-chondrophytic' spurs, compared with trochlear hyaline articular cartilages, incorporated less 35S-SO4 and 3H-glycine and synthesized less extractable high density 35S-PG monomers able to interact with exogenous HA. Their 35S-GAG side chains were more heterogeneous and segregated into three distinct peaks as shown by Sephadex G-200 column chromatography. The results of the present studies demonstrate that, in response to the implant, there was an increase in the biosynthetic capacity of chondrocytes which synthetized larger PG monomers able to interact wih HA.     

Osteoarthritis of the Knee in the Rabbit Produced by Immobilization:: Attempts to Achieve a Reproducible Model for Studies on Pathogenesis and Therapy

In order to obtain a reproducible experimental model of osteoarthritis a method of immobilizing the rabbit's knee in extension by means of a plastic splint was developed. The right knees of the rabbits were immobilized for periods varying from 4 days to 24 weeks. With the left knees as controls the knees were studied in a variety of ways among these being radiography (126 rabbits), histological sections stained with Alcian Blue (88 rabbits), analysis of ³⁵S-sulphate uptake (22 rabbits) and ³⁵S-autoradiography (6 rabbits). In 27 rabbits the regaining of mobility after immobilization was studied. After 5-6 weeks of immobilization most of the knees showed moderate or severe changes including loss of articular cartilage and osteophyte formation. Immobilization of the rabbit's knee by this method provokes a fairly easily reproducible type of degenerative joint disease showing similarities to advanced osteoarthritis as seen in humans.