Intrathecal liproxstatin-1 delivery inhibits ferroptosis and attenuates mechanical and thermal hypersensitivities in rats with complete Freund's adjuvant-induced inflammatory pain

Previous studies have confirmed the relationship between iron-dependent ferroptosis and a peripheral nerve injury-induced neuropathic pain model.However,the role of fe rroptosis in inflammatory pain remains inconclusive.Therefore,we aimed to explore whether ferroptosis in the spinal cord and do rsal root ganglion contributes to complete Freund's adjuvant(CFA)-induced painful behaviors in rats.Our results revealed that various biochemical and morphological changes were associated with ferroptosis in the spinal cord and dorsal root ganglion tissues of CFA rats.These changes included iron overload,enhanced lipid peroxidation,disorders of anti-acyl-coenzyme A synthetase long-chain family member 4 and glutathione peroxidase 4 levels,and abnormal morphological changes in mitochondria.Intrathecal treatment of liproxstatin-1(a ferroptosis inhibitor)reve rsed these ferroptosis-related changes and alleviated mechanical and thermal hype rsensitivities in CFA rats.Our study demonstrated the occurrence of fe rroptosis in the spinal cord and do rsal root ganglion tissues in a rodent model of inflammatory pain and indicated that intrathecal administration of fe rroptosis inhibitors,such as liproxstatin-1,is a potential therapeutic strategy for treating inflammatory pain.

Anti-inflammatory effects of Solanum procumbens on a low dose complete Freund's adjuvant-induced arthritis rat model

Objective:To investigate the anti-inflammatory and analgesic effects of Solanum procumbens on complete Freund’s adjuvantinduced arthritis rat models.Methods:We isolated and identified five compounds in the ethanolsoluble Solanum procumbens extract(SP)with anti-inflammatory effects,including ursolic acid,β-sitosterol,hexadecanoic acid,cisvaccenic acid,and vanillic acid.Additionally,we investigated the anti-inflammatory effects of SP on rheumatoid arthritis symptoms,including paw volumes,local temperatures,withdrawal latency,and mechanical withdrawal threshold at the hind paw and white blood cell(WBC)number from complete Freund’s adjuvant-induced arthritis rat models.Results:We have successfully established a complete Freund’s adjuvant-induced arthritis rat model at a low dose(1 mg/mL).SP extract significantly reduced paw volumes(P<0.05),prolonged withdrawal latencies(P<0.05),decreased local temperature,and increased the mechanical withdrawal threshold(P<0.05),but only SP extract at the dose of 300 mg/kg significantly decreased WBC numbers.Conclusions:SP extract could be a potential medication candidate with anti-inflammatory effects for arthritis,but it requires further investigation into the mechanism of the SP and its effectiveness on other models as well as clinical trials.

Meibomian gland dysfunction model induced with complete Freund’s adjuvant in C57BL/6 mice

AIM:To establish a new inflammatory animal model of meibomian gland dysfunction(MGD)in C57BL/6 mice.METHODS:C57BL/6 mice were randomly divided into complete Freund’s adjuvant(CFA)group(14 animals,14 eyes),naphthazolin hydrochloride(NH)group(14 animals,14 eyes)and control group(14 animals,14 eyes).In CFA group,CFA was used in eyelid conjunctiva injection;in NH group,NH eye drops were used twice a day;control group was injected with equal dose of saline at the same time point and same site with animals in CFA group.The meibomian gland orifices score(MGOS)was evaluated on a scale of 0 to 3 in the middle five meibomian gland orifices of the upper and lower eyelid using slit lamp.After the successful induction of each animal model,intense pulsed light(IPL)was introduced on each mouse in CFA and NH group.Oil red O(ORO),hematoxylin and eosin(H&E)staining were performed before and after successful induction of CFA,NH and control group.RESULTS:At 12wk after CFA injection,inflammatory cell infiltration and fiber necrosis was observed,with acinar density and duct dilatation significantly lower compared with control group.In NH group,the meibomian gland acini were relatively smaller and deformed compared with control group,the number of meibomian gland acini was also slightly lower.No inflammatory cell or fiber necrosis was observed in NH group.After three times of IPL treatment(5/10 mice in each group,and the other 5 mice served as non-IPL control),MGOS was significantly lower in IPL-treated mice in NH group(P<0.01).After three times of IPL treatment,the MGOS of NH group was significantly lower than that in the CFA group(P<0.01).CONCLUSION:We develop a novel animal model that studies the role of inflammation in the development of MGD and IPL treatment.This model indicates that persistent inflammatory state may be the cause of MGD and weaken the therapeutic effect of IPL.

机械穿刺结合肿瘤坏死因子α及完全弗氏佐剂构建大鼠椎间盘源性腰痛模型

背景:椎间盘退变是导致下腰痛的重要原因。目前国内外对于椎间盘退变的造模方法众多,但缺乏一个针对椎间盘退变引起下腰痛的模型。目的:对比机械穿刺结合肿瘤坏死因子α+完全弗氏佐剂注射与传统仅机械穿刺下建立的大鼠椎间盘源性腰痛模型的效果。方法:将18只成年SD雄性大鼠,按照随机数字表法分3组,每组6只,空白组不进行任何操作;对照组大鼠L_(4-5)椎间盘建立传统机械穿刺椎间盘退变模型;实验组在机械穿刺基础上,以微量注射器向L_(4-5)椎间盘内注入肿瘤坏死因子α+完全弗氏佐剂建立机械穿刺结合炎症因子诱导的椎间盘退变模型。术后4周,热板法测大鼠痛阈值用于评估椎间盘退变大鼠对热伤害感受;MRI观察各组大鼠椎间盘退变情况;ELISA法检测血清肿瘤坏死因子α、白细胞介素1β、白细胞介素6、前列腺素E2水平;苏木精-伊红和番红O固绿染色观察椎间盘形态变化。结果与结论:①在疼痛上,与对照组相比,实验组大鼠的行为学痛阈值持续降低,血清学各炎症因子指标显著增高;②在形态上,与对照组相比,实验组大鼠的MRI L_(4-5)髓核信号完全消失;组织病理学染色显示:对照组大鼠髓核完整,可见较多脊索细胞,部分纤维环破裂;实验组大鼠的脊索细胞稀少,髓核与纤维环结构紊乱,边界消失;③结论:大鼠椎间盘机械穿刺结合肿瘤坏死因子α+完全弗氏佐剂注射可成功建立椎间盘源性腰痛模型,该操作方法简单、经济,模型退变及腰痛明显,明显缩短椎间盘退变成模周期,可作为研究椎间盘源性腰痛模型的参考。

经完全性手术切除的Ⅱ~ⅢA期EGFR野生型NSCLC术后辅助化疗中预防应用PEG-rhG-CSF的临床获益分析

目的探究经完全性手术切除的Ⅱ~ⅢA期表皮生长因子受体(EGFR)野生型非小细胞肺癌(NSCLC)术后辅助化疗中预防应用聚乙二醇化重组人粒细胞集落刺激因子(PEG-rhG-CSF)的临床获益。方法前瞻性选取2019年2月至2021年2月广东医科大学附属医院经完全性手术切除的Ⅱ~ⅢA期EGFR野生型NSCLC术后辅助化疗患者90例作为研究对象,根据随机数字表法将患者分为观察组与对照组,每组各45例。对照组预防性应用重组人粒细胞(rhG-CSF),观察组预防性应用PEG-rhG-CSF。对患者随访3个月,比较两组治疗前及治疗1、3、5、10 d后的白细胞计数、中性粒细胞计数水平及白细胞减少症、中性粒细胞减少症发生情况,并记录患者随访期内化疗后的不良反应情况。结果两组患者在治疗前及治疗1、3、5 d后的白细胞计数水平比较,差异均无统计学意义(P>0.05);治疗10 d后,观察组患者的白细胞计数水平为(5.65±1.11)×10^(9)/L,高于对照组[(4.66±1.08)×10^(9)/L],差异有统计学意义(P<0.05)。两组患者在治疗前及治疗1、3、5 d后的中性粒细胞计数水平比较,差异均无统计学意义(P>0.05);治疗10 d后,观察组患者的中性粒细胞计数水平为(3.61±1.51)×10^(9)/L,高于对照组[(2.65±1.46)×10^(9)/L],差异有统计学意义(P<0.05)。观察组患者2、3级白细胞减少症和中性粒细胞减少症发生率分别为15.56%、11.11%,均低于对照组(35.56%、42.22%),差异均有统计学意义(P<0.05)。两组患者在骨骼肌疼痛、疲劳、感染、发热、胃肠反应等方面比较,差异均无统计学意义(P>0.05)。结论对经完全性手术切除后Ⅱ~ⅢA期EGFR野生型NSCLC患者,术后辅助化疗中预防应用PEG-rhG-CSF可以提高白细胞计数和中性粒细胞计数水平,并减少白细胞减少症和中性粒细胞减少症的发生,有助于改善患者的免疫功能和预后。

Human umbilical cord mesenchymal stem cells as treatment of adjuvant rheumatoid arthritis in a rat model

AIM:To investigate the effect of human umbilical cord stem cells,both mesenchymal and hematopoietic(CD34+),in the treatment of arthritis.METHODS:Mesenchymal stem cells(MSCs) and hematopoietic(CD34+) stem cells(HSC) were isolated from human umbilical cord blood obtained from the umbilical cord of healthy pregnant donors undergoing fullterm normal vaginal delivery.MSC,HSC,methotrexate(MTX) and sterile saline were injected intra-articularly into the rat hindpaw with complete freunds adjuvant(CFA) induced arthritis after the onset of disease(day 34),when arthritis had become well established(arthritis score ≥ 2).Arthritic indices were evaluated and the levels of interleukin(IL)-1,tumor necrosis factor(TNF)-α and interferon(IFN)-γ and anti-inflammatory cytokine IL-10 in serum were determined using enzyme-linked immunosorbent assay.Animals of all groups were sacrificed 34 d after beginning treatment,except positive control(PC) which was sacrificed at 10,21 and 34 d for microscopic observation of disease progression.We used hematoxylin,eosin and Masson's trichrome stains for histopathological examination of cartilage and synovium.RESULTS:The mean arthritis scores were similar in all groups at 12 and 34 d post immunization,with no statistical significant difference.Upon the injection of stem cells(hematopoietic and mesenchymal),the overall arthritis signs were significantly improved around 21 d after receiving the injection and totally disappeared at day 34 post treatment in MSC group.Mean hindpaw diameter(mm) in the MSC rats was about half that of the PC and MTX groups(P = 0.007 and P = 0.021,respectively) and 0.6 mm less than the HSC group(P = 0.047),as indicated by paw swelling.Associated with these findings,serum levels of TNF-α,IFN-γ and IL-1 decreased significantly in HSC and MSC groups compared to PC and MTX groups(P < 0.05),while the expression of IL-10 was increased.Histopathological examination with H and E stain revealed that the MTX treated group showed significant reduction of leucocytic infiltrate and hypertrophy of the synovial tissue with moderate obliteration of the joint cavity.Stem cells treated groups(both hematopoietic CD34+ and mesenchymal),showed significant reduction in leucocytic infiltrate and hypertrophy of the synovial tissue with mild obliteration of the joint cavity.With Masson's trichrome,stain sections from the PC group showed evidence of vascular edema of almost all vessels within the synovium in nearly all arthritic rats.Vacuoles were also visible in the outer vessel wall.The vessel became hemorrhagic and finally necrotic.In addition,there was extensive fibrosis completely obliterating the joint cavity.The mean color area percentage of collagen in this group was 0.324 ± 0.096,which was significantly increased when compared to the negative control group.The mean color area percentage of collagen in hematopoietic CD34+ and mesenchymal groups was 0.176 ± 0.0137 and 0.174 ± 0.0197 respectively,which showed a marked decrement compared to the PC group,denoting a mild increase in synovial tissue collagen fibers.CONCLUSION:MSC enhance the efficacy of CFAinduced arthritis treatment,most likely through the modulation of the expression of cytokines and amelioration of pathological changes in joints.

The petroleum ether fraction of Celastrus paniculatus Willd. seeds demonstrates antiarthritic effect in adjuvant-induced arthritis in rats

Objective:Celastrus paniculatus(CP)Willd.is a plant indigenous to India with medicinal properties.It is used in the ayurvedic treatment of inflammatory ailments such as rheumatoid arthritis.The present study was designed to investigate the therapeutic efficacy of the petroleum ether fraction(PCP)obtained from CP seeds on adjuvant-induced arthritis in rats.Methods:Arthritis was induced in SpragueeDawley rats by immunization with Freund’s complete adjuvant(FCA).Arthritis severity was evaluated by arthritis score,paw volume,tibiotarsal joint thickness,body weight,dorsal flexion pain,motility test,stair climbing ability and index of thymus and spleen.Moreover,histopathology of knee joints supported by haematological analysis was used to assess the anti-arthritic action of PCP.The levels of superoxide dismutase(SOD),glutathione(GSH),catalase(CAT),nitric oxide(NO)and malondialdehyde(MDA)in liver were also assessed.Serum samples were collected for estimation of aspartate transaminase(AST),alanine transaminase(ALT)and alkaline phosphatase(ALP).In addition,cytokines such as tumour necrosis factor-alpha(TNF-a)and interleukin-6(IL-6)were estimated in plasma.Results:PCP significantly alleviated arthritic progression with regards to paw swelling,arthritic score,immune organ indices,hyperalgesic effect and body weight.This phenomenon was correlated with significant suppression of overproduction of inflammatory cytokines(TNFa and IL-6),oxidant stress markers(MDA and NO)and cellular enzyme(AST,ALT and ALP)levels versus arthritic rats without treatment.Moreover,PCP restored the decreased levels of SOD,CAT and GSH.Conclusion:Our results suggest that the anti-arthritic properties of PCP may be due to immunosuppressive effects,cytokine regulation,antioxidant effects and bone-protective activities.

EFFECT OF ELECTROACUPUNCTURE ON THE IMMUNOREACTIVITY OF FOCAL CUTANEOUS μ-OPIOID RECEPTOR IMMUNOREACTION-POSITIVE FIBERS IN ADJUVANT ARTHRITIC RATS

客观：学习 -opioid 的效果在煽动性的皮肤织物和 electroacupuncture (EA ) 的效果的受体(粗腐殖质) 表示在在助手的粗腐殖质积极纤维的热门免疫反应(红外) 关节炎(AA ) 老鼠。方法：48 只 SD 老鼠的一个总数被使随机化进控制(n =8 ) ，模型(n= 10 ) ， focus-side-EA (n = 10 ) ， non-acupoint-EA (n = 10 ) ，并且 healthy-side-EA (n = 10 ) 组。 AA 模型被完全的 Freund 的助手的下的注射建立( CFA ， 50 L )进左后部的 paw.EA ( 4-16 Hz ， 0.5-1.5 V )为 30 min 在焦点或健康方面和 non-acupoints 上被用于“ Huantiao ”( GB 30 )和“ Yanglingquan ”( GB 34 )。使用的 Non-acupoints 是二个地点 5 公里到健康方面上的 GB 30 和 GB 34。在焦点的真皮、下的纸巾的热门粗腐殖质红外积极的纤维与 immunohistochemical 方法被染色。疼痛的严厉被把测试弄弯的脚(anklejoint ) 检测。结果：与模型组相比，“弯曲脚的测试” 20 在 第9 和 第11 天( P0.05 )并且在 non-acupoint-EA 组在 focus-side-EA 组显著地减少了在上 第8 ， 第9 并且在 CFA ( P 0.05 )的注射以后的 11thd ，都显示双边的 GB 30 和 GB 34 和 non-acupoints 的那 EA 能减轻疼痛。从第 13 天在上，没有重要差别在 3 个 EA 组和模型组(P0.05 ) 之中在“弯曲脚的测试”被发现分数。与控制组比较，在焦点织物的粗腐殖质红外积极的神经纤维的区域价值在在在模型的焦点的粗腐殖质红外积极的神经纤维的 .The 区域价值组织的 3 个 EA 组( P0.05 )是显著地更高的， 3 个 EA 组在控制组( P0.05 )比那高重要。与模型组相比，区域在组和 healthy-side-EA 组显著地增加了的 focus-side-EA (P 0.05 ) 粗腐殖质红外积极的纤维珍视；当在 non-acupoint-EA 组和 healthy-side-EA 组的粗腐殖质红外积极的纤维的那些在 focus-side-EA 是比那显著地低的时，组( P 0.05 )，和没有重要差别在粗腐殖质红外积极的纤维( P0.05 )的区域在模型组， healthy-side-EA 组和 non-acupoint-EA 组之中被发现，在焦点方面上显示 acupoints 的 EA 的更强壮的效果。结论：GB 30 和 GB 34 的 EA 能减轻煽动性的疼痛和起来调整在在 AA 老鼠的焦点的皮纸巾的粗腐殖质红外积极的纤维的表示，施加反煽动性、止痛的效果。

非小细胞肺癌术后多元化辅助治疗模式的研究进展

可手术切除Ⅱ-Ⅲa及Ⅲb(T3N2M0)期(IASLC/UICC第8版TNM分期)非小细胞肺癌(non-small cell lung cancer,NSCLC)推荐术后辅助化疗,但化疗毒性难以避免,且患者生存获益有限,仍具有较高的复发转移和死亡风险。在精准医疗时代,表皮生长因子受体酪氨酸激酶抑制剂(epidermal growth factor receptor tyrosine kinase inhibitor,EGFR-TKI)开启个体化辅助治疗的新篇章。本文将对NSCLC术后辅助化疗、放疗、EGFR-TKI靶向治疗、抗血管生成治疗、免疫检查点抑制剂治疗及中医药治疗进行综述,多元化辅助治疗模式的探索有望延缓术后复发转移,使NSCLC患者生存获益最大化。

乳癌新辅助化疗DCE-MRI预测病理完全缓解的应用价值

目的探讨动态增强磁共振成像(DCE-MRI)预测乳癌新辅助化疗(NAC)后病理完全缓解(pCR)的价值。方法回顾性选取2020年1月—2022年10月于我院接受NAC的乳癌病人108例,病人于NAC前和第2~3周期后各进行1次乳腺MR增强检查。根据术后Miller-Payne病理反应分级,将病人分为pCR组(36例)和non-pCR组(72例),比较两组临床病理资料及影像学特征。采用多因素Logistic回归分析筛选预测pCR的独立影响因素,并构建预测模型。采用受试者工作特征曲线的曲线下面积(AUC)评估模型预测pCR的效能。结果pCR组与non-pCR组比较,雌激素受体、孕激素受体、人表皮生长因子受体2(HER2)、组织学分级、肿瘤强化方式、肿瘤退缩方式、肿瘤最大径变化率(△D%)差异均有显著意义(χ^(2)=-3.12~29.79,t=6.09,P<0.05)。多因素Logistic回归分析显示,△D%(OR=1.046,95%CI=1.019~1.075,P<0.001)和HER2表达状态(OR=0.171,95%CI=0.038~0.758,P=0.020)是pCR的独立预测因素。其中△D%预测pCR的AUC最高(0.813),其次为HER2表达状态(0.660),二者联合诊断的AUC为0.854,灵敏度为80.6%,特异度为80.6%。结论乳癌NAC早期阶段DCE-MRI病灶△D%和HER2表达是预测NAC后pCR的重要指标,可以为临床治疗方案制定提供重要依据。

The analgesic effect of electroacupuncture in complete Freund's adjuvant rats:a meta analysis

Pain is a global problem and has been found sensitive to acupuncture. A large number of existing and newly published studies have demonstrated the analgesic effect of electroacupuncture(EA) in complete Freund's adjuvant(CFA) rat model, necessitating a new and comprehensive review. To evaluate the curative effect of EA on thermal hyperalgesia in CFA rats, and to explore parameters that influence treatment effects, studies of EA treatment on thermal hyperalgesia of CFA rat model were identified from nine databases up to June 17, 2017. Outcome measure was heat pain threshold. All the data were analyzed using Stata 14.0/MP software. 26 studies identified included 801 rats. The quality score of the studies ranged from 3 to 6, with a mean of 4.08. The effect of EA group was improvement in outcome compared with CFA group. In subgroup analyses, frequency 2, 100, 2/100 or 20-100 Hz. acupoint Huántiào(环跳GB30),Kūnlún(昆仑 BL60) +Xuánzhōng(悬钟 GB39), Zúsānlī(足三里 ST36) +BL60 or ST36+Sānyīnjiāo(三阴交SP6), retention time 20 or 30 min, treatment time for post-CFA injection immediately, the same day as CFA injection, post-CFA injection 24 or 48 h and interval 24 or 48 h all can improve the effect on CFA rat heat pain threshold. These results show that there is positive effective of EA in CFA-induced pain,different parameters have different effects on EA treatment of CFA heat hyperalgesia, however, because of the number of eligible researches and the high risk of bias, the evidence supporting this conclusion is limited.

网状蛋白1C水平与三阴性乳腺癌新辅助化疗疗效的相关性研究

目的探究三阴性乳腺癌(TNBC)患者外周血单个核细胞中网状蛋白1C(RTN-1C)表达水平与新辅助化疗疗效的关系。方法选取154例TNBC患者作为研究对象。收集所有受试者年龄、月经状态、病理类型、TNM分期、组织学分级和Ki-67表达>30%情况、肿瘤直径。抽取TNBC患者化疗前(T_(0))、化疗7 d(T_(1))、化疗14 d(T_(2))和化疗21 d(T_(3))的空腹肘正中静脉血,用Ficoll密度梯度离心法分离外周血中单个核细胞,蛋白质免疫印迹法检测单个核细胞中RTN-1C表达水平。受试者工作特征(ROC)曲线评价RTN-1C判断新辅助化疗疗效的效能,Logistic回归分析新辅助化疗疗效的风险因素,构建列线图回归模型预测新辅助化疗后病理学完全缓解,一致性指数(C-index)、校准曲线和决策曲线评估模型效能。结果根据新辅助化疗疗效分为完全缓解组(n=47)和未完全缓解组(n=107)。在T_(1)和T_(3)完全缓解组的RTN-1C均低于未完全缓解组(P<0.05);T_(0)、T_(1)和T_(2)的2组RTN-1C相对表达水平随时间进展均降低(P<0.01)。T_(3)-RTN-1C判断新辅助化疗后病理学完全缓解的ROC曲线下面积(AUC)高于T_(0)-RTN-1C、T_(1)-RTN-1C和T_(2)-RTN-1C(P<0.01)。Logistic回归分析结果显示T_(3)-RTN-1C>0.91(OR=12.178,95%CI:4.796~30.924)、N1期或N2期(OR=2.180,95%CI:1.100~4.322)、组织学分级Ⅲ级(OR=3.609,95%CI:1.453~8.969)是新辅助化疗后病理学未完全缓解的独立危险因素(P<0.05)。模型A(N分期、组织学分级和T_(3)-RTN-1C构建)的拟合曲线和理想曲线重合度较高,模型B(由N分期和组织学分级构建)的拟合曲线和理想曲线重合度较差。模型A、B的Cindex分别为0.866和0.772。当阈值概率大于0.40时,模型A判断新辅助化疗后病理学完全缓解的净收益高于模型B。结论N分期、组织学分级和T_(3)-RTN-1C构建的模型对判断新辅助化疗后病理学完全缓解有较高的区分度、精准度和临床应用价值。

足底注射完全弗氏佐剂致疼痛抑郁共病模型大鼠行为学及单胺类神经递质改变

目的观察足底注射完全弗氏佐剂对大鼠疼痛抑郁行为的影响及海马单胺类神经递质的变化,旨在建立慢性炎性疼痛抑郁共病动物模型。方法将16只8周龄SPF级健康雄性SD大鼠随机分为模型组与对照组,每组8只。模型组大鼠麻醉后通过左后足底注射完全弗氏佐剂100μL,建立慢性炎性疼痛抑郁共病模型大鼠,对照组注射同体积生理盐水。采用Von Frey纤维丝和热辐射刺激仪测量大鼠的疼痛阈值,采用旷场实验、悬尾实验、强迫游泳实验评价大鼠的抑郁行为,采用酶联免疫吸附实验检测大鼠海马组织的5-羟色胺、多巴胺、去甲肾上腺素的含量,采用苏木精-伊红(HE)染色观察大鼠海马区病理改变。结果与对照组相比,模型组大鼠机械缩足阈值及热缩足反射潜伏期在3、7、14 d明显降低(P<0.01);与对照组相比,模型组大鼠旷场实验运动总距离在7、14 d明显减少(P<0.01),中央区停留时长在14 d明显降低(P<0.01);与对照组相比,模型组大鼠悬尾实验不动时间在14 d明显增加(P<0.01);与对照组相比,模型组大鼠强迫游泳实验不动时间在7、14 d明显增加(P<0.05,P<0.01);与对照组相比,模型组大鼠海马中5-羟色胺、多巴胺、去甲肾上腺素含量明显减少(P<0.01)。与对照组相比,模型组大鼠海马组织病理发生改变,神经元形态不规则,排列松散、紊乱,细胞间隙增大,部分细胞核模糊,出现部分神经元固缩、凋亡。结论100μL完全弗氏佐剂足底注射可引起大鼠痛觉过敏、抑郁样行为改变及海马单胺类神经递质明显降低,造成海马病理形态变化,可有效模拟疼痛抑郁共病的表现,是一种可用于慢性炎性疼痛抑郁共病病理机制研究的实验模型。

β-制动蛋白2在炎性痛小鼠脊髓背角的表达及作用

目的:通过建立不同类型的炎性痛模型,观察β-制动蛋白2(Arrb2)在炎性痛小鼠疼痛发展过程中的作用。方法:在WT小鼠和Arrb2^(-/-)小鼠建立由卡拉胶(carrageenan)诱导急性炎性痛模型和弗氏完全佐剂(CFA)诱导慢性炎性痛模型,通过von Frey纤维丝法测定不同时间点的机械缩足反应阈值(PWT);利用Western Blot技术观察Arrb2在急性痛和慢性痛中的表达;在脊髓背角采用鞘内给药的方式,通过von Frey纤维丝法检测小鼠给予μ受体选择性激动剂(DAMGO)后短期和长期的PWT。结果:与WT小鼠相比,Arrb2^(-/-)小鼠在急性痛状态下痛敏恢复程度较慢,而在慢性痛早期表现出显著的抑痛作用。Western Blot结果显示:在急性痛模型下,脊髓背角Arrb2蛋白表达无明显变化,而在慢性痛模型下,Arrb2蛋白表达下降。最后,鞘内注射DAMGO实验表明,在急性痛模型下,敲除Arrb2后鞘内注射DAMGO短期无显著作用,后期可诱发吗啡耐受样痛敏;在慢性痛状态下,敲除Arrb2后鞘内注射DAMGO则在短期和长期均表现出镇痛作用的显著增强。结论:Arrb2在不同类型的炎性痛中表达和作用均有所不同。

紫草素对完全弗氏佐剂诱导的类风湿性关节炎小鼠模型的治疗作用

为探讨紫草素对类风湿关节炎(RA)的影响,将ICR小鼠随机分为4组:正常对照组和3个试验组(模型组、阳性对照组、紫草素组)。造模15 d后给药治疗60 d,阳性对照组灌胃美洛昔康[50 mg/(kg·d)],紫草素组灌胃紫草素[2 mg/(kg·d)],正常对照组和模型组灌胃等体积的生理盐水,分离小鼠右足踝关节部位,对滑膜炎症程度(0-3级)和软骨破坏程度(0-4级)以及骨浸润进行评估,分离小鼠胸腺和脾脏,计算胸腺指数和脾脏指数,并对各组小鼠进行组织病理学的评价和部分血清生化指标(致炎因子和抗氧化因子)的分析测定。结果表明,紫草素能显著改善小鼠的关节红肿症状。紫草素组中一氧化氮(NO)、肿瘤因子α(TNF-α)、白细胞介素6(IL-6)和白细胞介素1β(IL-1β)含量显著低于AA模型组,血清中丙二醛(MDA)含量显著低于AA模型组,超氧化物歧化酶(SOD)酶活力显著高于AA模型组,差异有统计学意义(P<0.05或P<0.01)。紫草素可以通过降低炎症因子水平,提高机体抗氧化能力从而减轻RA的炎症反应。

乳剂类免疫佐剂研究进展

乳剂类免疫佐剂又称为油类佐剂,是将抗原包裹在各种油相中,以实现为抗原提供贮存库,并通过将抗原靶向递呈给免疫细胞以增强机体对抗原的特异性免疫应答反应的一类免疫佐剂。自1916年首次报道乳剂类佐剂的应用至今,一百多年来乳剂类佐剂已成为疫苗研制与开发中不可缺少的重要因素,为应对人类和动物各种传染病提供更安全有效的疫苗提供了物质保障。随着现代疫苗学不断的发展,新型疫苗的研制和开发对佐剂提出了更高的要求,了解佐剂的组成成分、有效性和安全性的免疫作用机理是佐剂应用于疫苗研制和开发的前提。因此本文选择了白油佐剂、完全弗氏佐剂(Complete Freund’s Adjuvant,CFA)、不完全弗氏佐剂(Incomplete Freund’s adjuvant,IFA)、MF59®、AS(Adjuvant systems,AS)系列和Montanides系列等目前常用和商品化的乳剂类免疫佐剂,对其佐剂组成成分、免疫作用机理和临床应用情况等进行综述,以期为乳剂类免疫佐剂的临床使用提供一定的理论参考。

电针对佐剂性关节炎大鼠病灶局部皮肤组织IL-1β、IL-1ra mRNA表达的影响

目的 :观察电针对单侧佐剂性关节炎大鼠的疼痛试验评分、炎症灶局部病理反应 ,以及病灶局部皮肤组织IL 1β、IL 1ramRNA表达的影响。方法 :通过将完全弗氏佐剂 50 μL注入SD大鼠左后肢外踝关节皮下制造单侧佐剂性关节炎疼痛模型 ,电针“环跳”穴、“阳陵泉”穴 (刺激参数为0 .5～ 1.5V ,4～ 16Hz,3 0min) ,应用跖、背屈关节评分连续观察大鼠 7d内的痛反应曲线 ,应用HE组织化学染色方法观察病灶局部皮肤组织炎性病理反应 ,应用RT PCR技术检测IL 1β、IL 1ramRNA表达情况。结果 :电针可明显减少炎症痛大鼠的疼痛评分 ,抑制佐剂性关节炎大鼠病灶局部炎性病理反应 ,并抑制病灶局部皮肤组织IL 1βmRNA表达 ,促进IL 1ramRNA表达 ,使IL 1ra/IL 1β比值上升 (P <0 .0 1)。结论 :电针可能通过调节炎症灶局部致炎细胞因子及抗炎细胞因子之间的平衡 ,从根本上解除局部病灶免疫细胞的激活状态 ,达到扶正祛邪、平衡阴阳的调控作用 ,从外周途径缓解疼痛。

丹皮多糖-2b对大鼠糖尿病性白内障防治作用研究

目的:观察丹皮多糖-2b对大鼠糖尿病性白内障的防治作用。方法:采用链尿佐菌素(STZ)联合完全弗氏佐剂(CFA)诱导大鼠糖尿病性白内障,裂隙灯观察各组晶体出现初次混浊的时间,比较晶体的混浊程度。比色法测定各组血清及晶体中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-pX)、过氧化氢酶(CAT)活力和丙二醛(MDA)含量以及晶体中Na+-K+-ATP酶活力、高分子质量蛋白和不溶性蛋白含量。结果:裂隙灯检查结果显示PSM2b有明显的延缓糖尿病性白内障的发生,减轻晶体混浊程度的作用。与模型组比,PSM2b各剂量组均能明显降低大鼠血清及晶体中升高的MDA水平,中、高剂量组可升高血清及晶体中SOD,GSH-pX,CAT水平,上调晶体Na+-K+-ATP酶活力,且剂量与效果呈正相关。PSM2b中、高剂量组对高分子质量蛋白及不溶性蛋白含量亦有较好调控作用。结论:PSM2b对大鼠糖尿病性白内障具有显著的防治作用。

链佐星-弗氏完全佐剂制作大鼠糖尿病眼病并发症模型

目的 探讨建立适合于糖尿病眼病并发症及其治疗学研究的病理模型制作方法。方法 雄性Wistar大鼠ip链佐星3 0mg·kg- 1 ,d 2每只ip弗氏完全佐剂(CFA) 0 .1mL ,链佐星和CFA均每周1次,连续3周,血糖稳定升高后给予高脂饲料喂养。酶反应荧光法测定晶状体内山梨醇含量,视网膜铺片以OPTMAS软件分析内皮细胞/周细胞(E/C)比值及微血管密度,普通石蜡切片观察视网膜病理变化,电镜观察晶状体病理变化。结果 造模成功后血糖保持1 6mmol·L- 1 以上达1 2周,模型动物85 %出现明显白内障,晶状体内山梨醇含量明显升高,视网膜毛细血管密度显著增加,E/C比值升高,视网膜、晶状体病理变化明显。结论此模型与临床糖尿病眼病并发症有一定相似之处。