不同剂量弗氏完全佐剂对类风湿关节炎模型大鼠复制的影响

目的观察不同剂量的弗氏完全佐剂对类风湿关节炎(Rheumatoid arthritis,RA)模型复制的影响,探讨RA模型复制中弗氏完全佐剂的最佳注射量,以期为RA的实验研究提供较好的模型复制方法。方法采用"病证结合"复合佐剂性关节炎模型复制方法,即"风、寒、湿"环境因素+弗氏完全佐剂的模型复制方法,观察不同注射量的弗氏完全佐剂对RA模型复制的影响。结果 0.1mL模型复制组有自愈的趋势;0.15mL模型复制组大鼠体质量、跖围改变呈进行性下降趋势,X线可见骨质破坏情况较贴近临床患者骨质的改变;0.2mL模型复制组骨质呈粉碎性破坏。结论 0.15mL注射量可能是RA模型复制中的最佳注射量。     

链佐星所致糖尿病大鼠主动脉内皮依据舒张反应损伤的特征

AIM: To study whether impaired endothelium-dependent relaxation (EDR) in early diabetic mellitus in response to different receptor-mediated and nonreceptor-mediated vasodilators ran parallel and its possible mechanism. METHODS: Isometric tension recording in aortic rings from streptozotocin (Str)-induced diabetic and age-matched nondiabetic rats. RESULTS: EDR induced by receptor agonist acetylcholine (ACh), histamine (His) or bradykinin (BK) were all significantly reduced in diabetic rings compared with control rings, whereas nonreceptor agonist calcimycin-induced EDR was well reserved in diabetic rings [IC50 control: (0.13 +/- 0.07) mumol.L-1 diabetic: (0.14 +/- 0.06) mumol.L-1, P > 0.05, n = 7]. Cyclopiazonic acid (CPA) which also is a nonreceptor mediated endothelium-dependent vasorelaxant and cells' capacitative Ca2+ entry stimulant, failed to trigger EDR in diabetic rings. Pretreatment with N omega-nitro-L-arginine methylester (L-NAME, 0.3 mmol.L-1) not only abolished all of the EDR elicited by above mentioned vasodilators in either of diabetic or control rings, but also leveled responses triggered by each of the agonists between diabetic and control rings. Upon the maximal EDR induced by ACh (1 mol.L-1) or CPA (3 mumol.L-1) in phenylephrine (1 mumol.L-1) precontracted rings, calcimycin (1 mumol.L-1) further relaxed diabetic rings, but contracted control preparations. When endothelium was denuded, relaxation evoked by sodium nitroprusside and contractions triggered by CPA or His were all identical between diabetic and control rings. CONCLUSION: Receptor agonists but not nonreceptor agonists-induced EDR are commonly impaired in 4-wk Str-induced diabetic rat aorta, and this defective effect is attributable to the low formation of EDRF/NO which is related to impaired capacitative Ca2+ entry pathway in endothelium.     

星形胶质细胞在完全弗氏佐剂诱导的炎性痛中的作用

目的 探讨脊髓背角星形胶质细胞在完全弗氏佐剂(complete Freund's adjuvant,CFA)诱导的炎性痛中的作用.方法 雄性SD大鼠随机分为3组(6只/组):CFA组(大鼠左后肢足底注射CFA诱导炎性痛,同时行鞘内置管术,术后6 d鞘内给予生理盐水);L-α-aminoadipate(LAA)组(处理方式同CFA组,但术后6 d鞘内给予LAA);对照组(处理方式同CFA组,但足底注射生理盐水).术前2 d及术后7 d检测各组机械痛和热痛阈值,术后7 d以Real-time PCR法检测脊髓背角GFAP mRNA变化.结果 术前2 d各组大鼠左后肢足底机械痛及热痛阈值无明显差异(P> 0.05).术后7 d,CFA组大鼠脊髓背角GFAP mRNA较对照组明显增高(P<0.01),左后肢足底机械痛及热痛阈值较对照组明显降低(P<0.01);与CFA组相比,LAA组大鼠脊髓背角GFAP mRNA明显降低(P<0.01),左后肢足底机械痛及热痛阈值均明显增高(P<0.01).结论 LAA特异性抑制脊髓背角星形胶质细胞活性,缓解了CFA大鼠炎性痛,提示脊髓背角星形胶质细胞活化是CFA诱导炎性痛的重要机制.     

熟地黄对弗氏佐剂诱导大鼠足趾炎症的影响

目的熟地黄常出现于治疗类风湿性关节炎的方剂中,本文用完全弗氏佐剂诱导大鼠足趾炎症,旨在验证其是否具有抗炎作用。方法于大鼠右后足趾皮内注射完全弗氏佐剂100μL。致炎后第19天,灌胃给予高、中、低剂量的熟地黄[0.625,1.250,2.500g/(kg.体重)],10d,以地塞米松为阳性药物,记录大鼠体重和足趾体积变化;第29天,测定脏器、血液参数,取足趾肉垫切片观察。结果模型组大鼠足趾持续肿胀,淋巴细胞浸润,胸腺和脾脏指数下降,血小板显著减少,其它指标与对照组差异不显著。熟地黄未显示抗炎作用,但可使胸腺和脾脏指数,以及血小板计数恢复正常,使肌酐水平显著上升。地塞米松抗炎作用显著,但同时存在多项毒副作用。结论熟地黄在治疗类风湿性关节炎的方剂中主要发挥辅助作用。     

在弗氏链霉菌中天冬氨酸转移酶和泰洛星生物合成

泰洛星(Tn)是由弗氏链霉菌所产生的大环内酯类抗生素,泰洛星的化学结构已被阐明,即由前泰乐内酯(protylonolide)连接3个糖所组成.它的生物合成可分为两部分:一是从单元分子(丙酸、乙酸和丁酸)形成的前泰乐内酯,另一是关于三糖的合成及其依次连接到前泰乐内酯上,据报道.单元分子由氨基酸转化成相应的羧酸来提供.而     

弗氏佐剂致炎大鼠脊髓背角Fos神经元的表达

目的探讨完全弗氏佐剂（CFA）致炎性痛大鼠Fos神经元（Fos-LIN）在脊髓背角痛觉传导通路中的表达及意义。方法①16只SD大鼠随机均分为两组,于右后肢踝关节外侧皮下分别注射0.9%生理盐水（NS）或CFA50μl,观察注射前后大鼠热缩足反射潜伏期TWL的变化;②48只大鼠随机平分为两组后分别注射NS和CFA（剂量和注射部位同前）以免疫组化法检测脊髓背角Fos-LIN表达。结果①大鼠注射CFA后TWL逐渐降低,12h时为最低点,持续3d后逐渐恢复,14d时基本恢复至基础水平;②NS组脊髓背角Fos-LIN散在出现,而CFA组中Fos-LIN1h时最先出现在大鼠右侧脊髓背角I-II层,4h时出现在V-VI层的数目逐渐增多,12h时为脊髓全层出现最多,至14d时在背角各层少见,且同组左侧脊髓各层Fos-LIN在1h-14d均比较少见。结论50μlCFA能诱导大鼠产生为期2周的炎性痛觉过敏。炎症期间脊髓背角Fos神经元的持续表达与外周痛觉信号的传导和中枢的调控有关。     

牛膝多糖对链佐星诱导性糖尿病小鼠的保护作用(英文)

目的研究牛膝多糖(ABP)对链佐星诱导性糖尿病小鼠的保护作用。方法将雄性ICR小鼠分为正常对照组、糖尿病模型组、ABP50和100mg.kg-1(ip,每天1次,共15d)治疗组。分别于给药前,给药8和15d时用血糖测量仪检测血糖水平,并进行血糖耐受试验。末次给药第2天测量小鼠体重,处死,测量心、肝、脾和肾脏器官重量,同时制备血清,采用放射免疫分析试剂盒检测血清胰岛素水平,比色法测定血清甘油三酯(TG)、总胆固醇(TC)、钙(Ca)和磷(P)含量及谷草转氨酶(GOT)、谷丙转氨酶(GPT)和碱性磷酸酶(ALP)活性。采用ELISA检测血清瘦素、脂联素、肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)的浓度。结果与正常对照组相比,糖尿病模型组小鼠体重下降,血清胰岛素水平降低,血糖升高(P<0.05,P<0.01)。ABP50和100mg.kg-1在血糖耐受试验中能较好地调节血糖水平,其血糖在15d时分别比模型组下降了27.4%和16.3%(P<0.05,P<0.01)。ABP50mg.kg-1治疗组小鼠脾和肾脏指数,血糖、血清TG和TC浓度,GOT,GPT和ALP活性均显著低于糖尿病模型组(P<0.05),血清瘦素水平下降并接近正常水平,血清脂联素、TNF-α和IL-6水平均显著高于模型组(P<0.05,P<0.01);ABP100mg.kg-1对血清TC和TG水平及ALP和GPT活性无明显作用,对上述其他指标的作用与ABP50mg.kg-1相似。各组血清胰岛素浓度及Ca和P含量均无明显变化。结论ABP对链佐星诱导的糖尿病小鼠具有一定的保护作用。     

促肝细胞生长素对链佐星损伤性糖尿病大鼠的治疗作用

目的:研究促肝细胞生长素(PHGF)对链佐星损伤性糖尿病大鼠的治疗作用。方法:链佐星制作糖尿病大鼠模型,共60只大鼠,分为4组,每日分别给予PHGF(20mg·kg-1,iv)、胰岛素(2U·kg-1,im)、烟酰胺(1g·kg-1,ig)、生理盐水(1mL,iv)治疗3mo。葡萄糖氧化酶法测定大鼠血糖,放射免疫法测定胰岛素和C肽, 3T3细胞测定促肝细胞生长素体外活性,抗胰岛B细胞免疫组化染色法研究用药前后胰岛B细胞的病理变化,并测定B细胞数。结果:用药1mo, 4组大鼠血糖均降低,与治疗前相比,P<0. 01;PHGF组血糖下降(4. 6±s0. 9 )mmol·L-1,下降幅度大于其他3组,P<0. 05或P<0. 01。3mo后, 4组大鼠血胰岛素和C肽浓度增高,与治疗前相比,均P<0. 01;PHGF组升高最明显,与其他3 组比较差异有非常显著意义(P<0. 01)。并且PHGF组胰岛B细胞数每高倍镜(46±4)个,高于其他3组(P<0. 01 ); 5个批号的PHGF体外生物学活性均大于2. 0。结论:PHGF能改善胰岛B细胞分泌功能,具有使胰岛B细胞肥大、再生的作用。     

曲安奈德乳剂对佐剂型关节炎模型的治疗作用

目的采用佐剂型大鼠类风湿性关节炎模型评估曲安奈德棕榈酸酯脂质微球注射液对关节炎的疗效。方法采用弗氏佐剂给大鼠单侧造模后,分别于阳性对照组、受试药组和空白乳剂组大鼠双侧踝关节腔内一次性给予阳性对照药(曲安奈德注射液2.0 mg.kg-1)、受试药(曲安奈德棕榈酸酯注射液2.01、.0、0.5 mg.kg-1)及空白乳剂。每3 d检测1次各组双侧后足的肿胀度,直至给药后第12天采集所有动物血清,用ELISA法检测血清中IL-1β及TNF-α。并取原发侧踝关节组织及膝关节滑膜组织进行病理组织学观察及用免疫组化法测定MMP-3。结果曲安奈德棕榈酸酯注射液关节腔内注射可抑制佐剂型类风湿性关节炎模型大鼠的足肿胀度;降低血清IL-1β、TNF-α水平;减轻造模形成的各种组织病理学变化;抑制原发部位膝关节滑膜组织MMP-3水平。以上作用强度均与阳性药相当。结论曲安奈德棕榈酸酯注射液对佐剂型大鼠类风湿性关节炎模型有强大的治疗作用,且药效与阳性药曲安奈德注射液相当。     

空肠弯曲菌诱导系统性红斑狼疮样小鼠模型的优化

目的优化空肠弯曲菌（CJS131）诱导系统性红斑狼疮（SEE）样小鼠模型的方法。方法小鼠随机分为5组,即正常对照组、卡介苗（BCG）对照组、结核分枝杆菌H37Ra对照组、CJS131＋BCG模型组和CJS131＋H37Ra模型组。在第0天免疫,在第14、21、42天加强免疫,在第36、54、61天分批称其体质量后处死。检测小鼠免疫器官指数、血清抗核抗体水平、血清总IgG水平和肾组织病理损伤程度。结果不论采用含BCG还是H37Ra的弗氏完全佐剂（FCA）,给予CJS131免疫的小鼠均有SLE样综合征的表现：血清抗核抗体及总IgG水平升高,肾组织损伤。在动态检测中病变基本维持。结论CJS。在对模型组小鼠的免疫诱导中起主要作用,以BCG或H37Ra作为FCA中采用的菌株都可引起病变,通过加强免疫可维持小鼠长期病变,优化后的模型可用于SLE治疗性药物的筛选。     

银杏叶提取物对大鼠佐剂性关节炎的治疗作用与机制

目的:考察银杏叶提取物(EGb761)对佐剂性关节炎(AA)大鼠的治疗作用,初步探讨银杏叶提取物治疗AA的作用机制。方法:将大鼠随机分为正常对照组,模型组,雷公藤多苷组,EGb761低、中、高(50,100,200 mg.kg-1)剂量组。弗氏完全佐剂致AA后d 12,大鼠出现继发性炎症,分别灌胃给予雷公藤多苷和EGb761,连续16 d;在不同的时间点检测大鼠继发侧关节肿胀度;致炎d 28处死大鼠,酶联免疫法检测血清中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-4(IL-4)和白细胞介素-10(IL-10)含量;光学显微镜观察关节病理变化。结果:EGb761可明显减轻继发性AA大鼠足爪的肿胀度;降低大鼠血清中IL-1β和IL-6含量;升高大鼠血清中IL-4和IL-10含量。结论:EGb761对大鼠继发性AA具有显著的治疗作用,其作用机制可能与抑制促炎因子IL-1β和IL-6的产生和释放以及促进抑炎因子IL-4和IL-10的表达相关。     

Protective effects of fenofibrate and resveratrol in an aggressive model of rheumatoid arthritis in rats

Context Fibrates were reported to have anti-inflammatory effects while the naturally occurring polyphenol resveratrol was traditionally known as a potent antioxidant agent.

Objective The effects of fenofibrate and resveratrol were investigated on complete Freund’s adjuvant (CFA)-induced rheumatoid arthritis (RA) in adult female albino rats.

Materials and methods Rats were divided into a normal control group, an arthritis control group receiving CFA, two reference treatment groups receiving dexamesathone (1.5?mg/kg/day) and methotrexate (1?mg/kg/day), and two treatment groups receiving fenofibrate (100?mg/kg/day) and resveratrol (10?mg/kg/day) for seven consecutive days. Assessment of RA was performed by measuring serum rheumatoid factor (RF), matrix metalloprotinease-3 (MMP-3) and cartilage oligomeric matrix protein (COMP) as specific rheumatoid biomarkers, immunoglobulin G (IgG) and antinuclear antibody (ANA) as immunological biomarkers, tumour necrosis factor-alpha (TNF-α) and interleukin-10 (IL-10) as immunomodulatory cytokines, myeloperoxidase (MPO) and C-reactive protein (CRP) as inflammatory biomarkers and malondialdehyde (MDA) and glutathione (GSH) as oxidative stress biomarkers, supported by a histopathological study on joints and spleens.

Results Serum RF, MMP-3, COMP, IgG, ANA, TNF-α, MPO, CRP and MDA were decreased to about 36, 56, 66, 65, 9, 35, 24, 44 and 31% by fenofibrate, and to about 37, 59, 44, 70, 5, 30, 23, 33 and 28% by resveratrol treatments, respectively. Alternatively, serum IL-10 and GSH were significantly increased to about 215 and 251% by fenofibrate and to about 225 and 273% by resveratrol treatments, respectively.

Discussion and conclusion Fenofibrate and resveratrol protect against RA, possibly through their immunomodulatory, anti-inflammatory and antioxidant potential.     

A novel compound PTIQ protects the nigral dopaminergic neurones in an animal model of Parkinson's disease induced by MPTP

BACKGROUND AND PURPOSE

In Parkinson''s disease, the dopaminergic neurones in the substantia nigra undergo degeneration. While the exact mechanism for the degeneration is not completely understood, neuronal apoptosis and neuroinflammation are thought to be key contributors. We have recently established that MMP-3 plays crucial roles in dopaminergic cell death and microglial activation.

EXPERIMENTAL APPROACH

We tested the effects of 7-hydroxy-6-methoxy-2-propionyl-1,2,3,4-tetrahydroisoquinoline (PTIQ) on expression of MMP-3 and inflammatory molecules and dopaminergic cell death in vitro and in an animal model of Parkinson''s disease, and Parkinson''s disease-related motor deficits. The pharmacokinetic profile of PTIQ was also evaluated.

KEY RESULTS

PTIQ effectively suppressed the production of MMP-3 induced in response to cellular stress in the dopaminergic CATH.a cell line and prevented the resulting cell death. In BV-2 microglial cells activated with lipopolysaccharide, PTIQ down-regulated expression of MMP-3 along with IL-1β, TNF-α and cyclooxygenase-2 and blocked nuclear translocation of NF-κB. In the mouse model of Parkinson''s disease, induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), PTIQ attenuated the associated motor deficits, prevented neurodegeneration and suppressed microglial activation in the substantia nigra. Pharmacokinetic analysis showed it was relatively stable against liver microsomal enzymes, did not inhibit the cytochrome p450 isozymes or the hERG ion channel, exhibited no cytotoxicity on liver cells or lethality when administered at 1000 mg kg⁻¹ and entered the brain rather rapidly yielding a 28% brain:plasma ratio after i.p. injection.

CONCLUSIONS AND IMPLICATIONS

These results suggest PTIQ has potential as a candidate drug for disease-modifying therapy for Parkinson''s disease.     

Development of a human corneal epithelium model utilizing a collagen vitrigel membrane and the changes of its barrier function induced by exposing eye irritant chemicals

The brief TEER (trans-epithelial electrical resistance) assay after exposing chemicals to corneal epithelium in vivo is known as a suitable method for evaluating corneal irritancy and permeability quantitatively and continuously. A collagen vitrigel membrane we previously developed is a thin (about 20 μm thick) and transparent membrane composed of high density collagen fibrils equivalent to connective tissues in vivo, e.g. corneal Bowman’s membrane. To develop such a TEER assay system in vitro utilizing a human corneal epithelial model, HCE-T cells (a human corneal epithelial cell line) were cultured on the collagen vitrigel membrane substratum prepared in a Millicell chamber suitable for TEER measurement. Human corneal epithelium model possessing 5-6 cell layers sufficient for TEER assay was successfully reconstructed on the substratum in the Millicell chamber by culturing the cells in monolayer for 2 days and subsequently in air-liquid interface for 7 days. The exposure of chemicals to the model induced the time-dependent relative changes of TEER in response to the characteristic of each chemical within a few minutes. These results suggest that the TEER assay using the human corneal epithelial model is very useful for an ocular irritancy evaluation as an alternative to the Draize eye irritation test.     

Anthocyanins inhibit airway inflammation and hyperresponsiveness in a murine asthma model.

Asthma is a common chronic inflammatory disease regulated by coordination of T-helper cell type 2 (Th2) cytokines and inflammatory signal molecules. Additionally, oxidative stress may play an important role in airway inflammation such as eosinophilia, mucus hypersecretion, and airway hyperresponsiveness (AHR). In the present report, we investigated whether anthocyanins would reduce airway inflammation in a mouse asthma model immunized and challenged with ovalbumin (OVA). OVA inhalation elicited inflammatory responses characterized by eosinophilia and increased lipid hydroperoxide (LPO) in bronchoalveolar lavage (BAL) fluid, enhanced pause (Penh), increased glycoprotein and proliferating cell nuclear antigen (PCNA) expressions in mucus hypersecretion, and an increased expression of various cytokines and cyclooxygenase (COX) 2 in lung tissues. All parameters were attenuated in a dose-dependant manner by the administration of anthocyanins. These results suggest that anthocyanins may attenuate the development of asthma by downregulating Th2 cytokines, proinflammatory cytokines, and COX-2. Our findings suggest that anthocyanins have positive contributions as a dietary supplement for the prevention of asthma.