Therapeutic Efficacy of Quinapyramine Sulphate with Freund＇s Complete Adjuvant in Wistar Rats Infected with Trypanosoma Congolense

Therapeutic efficacy of QS （quinapyramine sulphate） and FCA （Freund＇s complete adjuvant） combination was studied. The aim of the study was to evaluate therapeutic efficacy of QS using FCA in Trypanosorna congolense infection. GrouPs treated with QS and FCA had parasite disappeared in peripheral circulation 2 days pi, relapse was observed one week later. Effect of treatment on rectal temperature shows no significance （p 〈 0.05）, normalization of rectal temperature occurred in QS and FCA treated groups （34.1℃） than untreated （42.8 ℃）, QS （37.4 ℃） and FCA （35.92 ℃） treated groups. Mean body weight was significant （p 〈 0.001） in QS and FCA, QS, and FCA groups. Packed cell volume and hemoglobin concentration for untreated groups were lower, but increased in QS, FCA, QS and FCA treated groups, indicating anemia amelioration. White blood cell counted in untreated, QS and FCA treated groups showed no significance （p 〈 0.05）, however, there was leukocytosis due to lymphocytosis in QS and FCA treated group （6.79 × 10^3/μl） compared with untreated and other groups. There was comparative decrease in serum liver enzymes in QS and FCA treated group than other groups. Therefore, QS at lower recommended dose with FCA may enhance efficacy of QS in trypanosomiasis.

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

Objective：To study the effect of μ-opioid receptor （MOR） expression in the inflammatory skin tissue and the effect of electroacupuncture （EA） on the topical immunoreaction （IR） of MOR positive fibers in adjuvant arthritic （AA） rats. Methods： A total of 48 SD rats were randomized into control （n = 8）, model （n = 10）, focus-side-EA （n = 10）, non-acupoint-EA （n = 10）, and healthy-side-EA （ n = 10） groups. AA model was established by subcutaneous injection of complete Freund＇s adjuvant （CFA, 50 μL） into the left hind paw. EA （4-16 Hz, 0.5-1.5 V） was applied to “Huantiao” （环跳 GB 30） and“Yanglingquan” （阳陵泉 GB 34） on the focus or healthy side and non-acupoints for 30 min. Non-acupoints used were the two sites 5 mm to GB 30 and GB 34 on the healthy side. The topical MOR IR-positive fibers in the dermal and subcutaneous tissues of the focus was stained with immunohistochemical method. The severity of pain was detected by foot （anklejoint）-bending test. Results： Compared with model group, the “foot-bending test” score decreased significantly in focus-side-EA group on the 9^th and 11^th day （P〈 0.05） and in non-acupoint-EA group on the 8^th, 9^th and 11thd after injection of CFA （P 〈 0.05）, indicating that EA of bilateral GB 30 and GB 34 and non-acupoints all can relieve pain. From the 13^th day on, no significant differences were found in “foot-bending test” scores among the 3 EA groups and model group （P 〉0.05）. In comparison with control group, the area values of MOR IR-positive nerve fibers in the focus tissue were significantly higher in 3 EA groups （P 〈 0.05）. The area values of MOR IR-positive nerve fibers in the focus in model group and 3 EA groups were significant higher than that in control group （P〈 0.05）. Compared with model group, the area values of MOR IR-positive fibers in focus-side-EA group and healthy-side-EA group increased significantly （P 〈 0.05）; while those of MOR IR-positive fibers in non-acupoint-EA group and healthy-side-EA group were significantly lower than that in focus-side-EA group （ P 〈 0.05 ）, and no significant differences were found among model group, healthy-side-EA group and non-acupoint-EA group in the area of MOR IR-positive fibers （P 〉0.05）, indicating a stronger effect of EA of acupoints on the focus side. Conclusion： EA of GB 30 and GB 34 can relieve inflammatory pain and up-regulate the expression of MOR IR-positive fibers in the focal skin tissues in AA rats, exerting anti-inflammatory and analgesic effects.

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.

Progresses of studies on acupuncture and pain signals transduction mechanisms in complete Freund's adjuvant arthritis

Advances of studies on the acupuncture and pain signal transduction mechanisms in complete Freud＇s adjuvant arthritis are reviewed from the three aspects, the first messenger of modulating pain signals and the related receptors, the second messenger of modulating pain signals and other factors possibly involved in modulation of pain signal transduction, etc. It is held that modulation of acupuncture for pain signals is a comprehensive course involved in multi-channels, multi-levels, multi-links, and in future, acupuncture analgesic mechanisms for Freud＇s adjuvant arthritis will be more deeply studied by use of more new techniques and new methods.

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.

佐剂FCA与Quil A对rGST-Sj32的免疫增强作用的比较

目的 比较弗氏完全佐剂 (FCA)与皂素的纯化物 Quil A的免疫增强作用 ,进一步提高重组抗原 r GST- Sj32的免疫效果。 方法 r GST- Sj32 +FCA或 Quil A免疫 NIH小鼠 3次 ,EL ISA法检测抗体水平 ,用减虫率及减卵率表示保护效果。 结果 与 PBS对照组比较 ,r GST- Sj32 +FCA免疫小鼠减虫率为 42 .9% ,每克肝卵 (L EPG)减少率 5 1.0 % ,每雌肝卵 (L EPF)减少率为 2 3.9% ,抗体滴度达 1∶ 2 5 6 0 0。r GST- Sj32 +Quil A免疫小鼠减虫率为 46 .0 % ,L EPG减少率39.4% ,L EPF减少率为 8.5 % ,抗体滴度达 1∶ 5 12 0 0。 结论 FCA和 Quil A均可增强 r GST- Sj32免疫小鼠的抗感染保护性免疫力 ;FCA亦可增强 r GST- Sj32诱导小鼠抗生殖免疫 ,Quil A无增强抗生殖免疫的作用。

两种佐剂对日本血吸虫重组抗原SjGST-Sj32保护效果的影响

目的比较弗氏完全佐剂（FCA）与单磷脂A（MPL）的免疫增强作用，为提高重组SjGST-Sj32（SjGST-Sj32）的疫苗效果提供依据。方法SjGST-Sj32加弗氏完全佐剂（SjGST-Sj32＋FCA）或单磷脂A（SjGST-Sj32＋MPL）免疫小鼠3次，ELISA法检测抗体水平，用减虫率及减卵率表示保护性作用。结果与PBS对照组相比，SjGST-Sj32加FCA或MPL，免疫BALBC／c小鼠减虫率分别为48．8％（P＜0．01）和44．1％（P＜0．01）；每克肝卵（LEPG）减少率分别为59．4％（P＜0．01）和46．2％（P＜0．01）；SjGST-Sj32＋FCA组抗体滴度达1：204800（P＜0．05），而SjGST-Sj32＋MPL组抗体滴度为1：51200（P＜0．05）。结论FCA和MPL均可以增强对SjGST-Sj32免疫小鼠的抗感染保护性免疫力；FCA也可增强SjGST-Sj32诱导小鼠抗生殖免疫，而MPL似不具明显增强SjGST-Sj32的抗生殖免疫作用。佐剂MPL在提高免疫小鼠体液免疫应答方面稍逊于经典佐剂FCA。

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。

完全弗氏佐剂致炎性痛大鼠脊髓背角Fos蛋白的表达

目的：探讨完全弗氏佐剂（CFA）所致炎性痛大鼠Fos蛋白在脊髓背角痛觉传导通路中的表达及意义。方法：64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，每组取8只观察注射前后热缩足反射潜伏期（TWL）的变化；余48只以免疫组化法检测脊髓背角Fos蛋白表达。结果：大鼠注射CFA后TWL逐渐降低，并在12h时达最低点（下降62％），持续3d后缓慢上升，14d时基本恢复至基础水平；NS组脊髓背角Fos蛋白散在表达，而CFA组中1h后Fos蛋白最先在大鼠右侧脊髓背角Ⅰ～Ⅱ层表达（P〈0.01），4h时Ⅴ～Ⅵ层蛋白表达逐渐增多（P〈0.01），12h时脊髓全层蛋白表达最多，至14d时背角各层少见Fos蛋白。结论：50μl CFA能诱导大鼠产生为期2周的炎性痛病程。炎症期间脊髓背角Fos蛋白的持续表达与外周痛觉信号的传导和中枢的调控有关。

Lewis大鼠实验性变态反应性脑脊髓炎动物模型的建立及百日咳疫苗的影响

目的建立可靠的Lewis大鼠实验性过敏性脑脊髓炎(experimental allergic encephalomyelitis,EAE)动物模型;方法采用Lewis大鼠脚垫皮下注射豚鼠全脊髓匀浆(spinal cords homogenate of guinea pigs,GPSCH)和含有卡介苗(bacille calmetteguerin,BCG)10mg/ml的完全福氏佐剂(complete Freund's adjuvant,CFA)诱导EAE,从临床症状评分、光镜、电镜下病理学改变对模型进行评价。结果Lewis大鼠EAE的发病率为9/10,潜伏期为12.6±1.01天。百日咳疫苗(bordetella pertussis vaccine,BPV)可提高EAE的发病率,缩短EAE的潜伏期(P<0.05),加重其临床症状和病理改变(P<0.05)。结论GPSCH和CFA免疫Lewis大鼠建立的EAE是成功可靠的;BPV对EAE有显著的促发作用,可诱导形成超急性EAE。

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。

The Prevention and Treatment of Polysaccharide from the Rhizomorph of Armillaria mellea on Diabetic Cataract in Rat

[Objective] To investigate the prevention and control effect of polysaccharide from the rhizomorph of Armillaria mellea by distilled water elution （AMP-1） on diabetic cataract in rats.[Methods] Streptozotocin （STZ）,combined with freund＇s adjuvant （CFA）,was used to induce diabetic cataract in rat,to observe the onset of lenticular turbidity by slit lamp and compare the turbid degree of eyes lens among different groups.The super oxide dismutase （SOD）,glutathione peroxidase （GSH-px） and malondialdehyde （MDA） in serum and eyes lens were measured by colorimetric method.[Results] The slit lamp examination results showed that AMP-1 obviously delayed the onset of diabetic cataract and reduced turbid degree of eyes lens.Compared with model group,AMP-1 at various doses should significantly reduce the level of MDA ascended in serum of rats.Medium-and high-dose groups could increase the levels of SOD and GSH-Px in serum,and the dose of AMP-1 was closely related to the effect.[Conclusion] AMP-1 played a prominent role in prevention and treatment of diabetic cataract in rats.

福氏完全佐剂包裹白细胞介素2和肿瘤坏死因子对小鼠S180肿瘤的抑制作用

本文报道一种新的免疫治疗方法。用福氏完全佐剂（Ｆｒｅｕｎｄ＇ｓ ｃｏｍｐｌｅｔｅ ａｄｊｕｖｅｎｔ，ＦＣＡ）包裹白细胞介素２（ｌｎｔｅｒｌｅｕｋｉｎ ２，ＩＬ－２）及／或肿瘤坏死因子（Ｔｕｍｏｒ Ｎｅｃｒｏｓｉｓ Ｆａｃｔｏｒ，ＴＮＦ）作了抗Ｓ１８Ｏ肉瘤的体内抗瘤实验。结果表明：ＩＬ－２及／或ＴＮＦ经佐剂包裹后，可明显减轻其毒副作用，并显著提高ＩＬ－２及／或ＴＮＦ的抗瘤作用。

完全弗氏佐剂雌性SD大鼠类风湿关节炎模型的建立及塞来昔布对其治疗效果

目的利用完全弗氏佐剂(CFA)建立雌性SD大鼠类风湿关节炎(RA)模型,并考察不同给药方式的塞来昔布对RA的治疗效果。方法将24只雌性SD大鼠随机分成正常对照组和低、中、高剂量模型组(n=6),低、中、高剂量模型组分别用0.05、0.1、0.2 mL CFA(10 mg/mL)于大鼠右后肢足跖部注射诱导构建RA模型,筛选出合适的剂量。另取24只雌性SD大鼠分为正常对照组、模型对照组、塞来昔布口服组、塞来昔布注射组(n=6),后3组利用最佳致炎剂量CFA建立RA模型。塞来昔布口服组给予市售塞来昔布胶囊,每天灌胃1次,每次20 mg;塞来昔布注射组给予用原料药配制的塞来昔布混悬液,每周于大鼠右后肢足跖部注射给药1次,剂量20 mg/kg。测量各组大鼠足容积,并进行临床评分。通过ELISA检测各组大鼠血清中3种炎症因子(IL-1β、IL-6、TNF-α)的浓度,取大鼠踝关节组织进行H-E染色以观察组织病变情况。结果在雌性SD大鼠中,足跖部注射0.1 mL CFA(10 mg/mL)可较好地模拟RA中度炎症反应,注射CFA后逐渐出现足爪红肿,全身临床评分及足部关节炎指数评分均有升高,炎症高峰期在免疫后19 d。塞来昔布口服组和注射组足容积较模型对照组减小(P均<0.05),全身临床评分及足部关节炎指数评分均较模型对照组降低(P均<0.01),血清IL-1β、IL-6、TNF-α浓度均较模型对照组降低(P均<0.05),踝关节组织病理改变均较模型对照组减轻;其中塞来昔布注射组的全身临床评分及足部关节炎指数评分低于口服组(P均<0.05),踝关节组织病理改变轻于口服组。结论足跖部注射0.1 mL CFA(10 mg/mL)可致SD雌性大鼠中度炎症反应,能较好地模拟RA的病理过程。每周经足跖部注射1次塞来昔布混悬液对模型大鼠RA的疗效优于每天口服1次塞来昔布。

Preparation and analysis of active rat model of rheumatoid arthritis with features of TCM toxic heat-stasis painful obstruction

Objective:To establish a collagen type II-induced rat model of rheumatoid arthritis(RA)presenting characteristics of the human form of the traditional Chinese syndrome pattern of toxic heat-stasis painful obstruction(bi zheng;arthromyodynia)as well as pathologic features of active RA.The Chinese herbal medicine Tengmei decoction was used to validate the animal model.Methods:Ninety specific pathogen free Sprague-Dawley rats were randomly divided into a normal group of 6 rats and a model group of 84 rats.To establish the rat model of collageninduced arthritis(CIA),bovine type II collagen in complete Freund’s adjuvant was injected into the model group rats as a priming dose(Day 0)and boosting dose(Day 9).Changes in arthritic index(AI)scores,including limb swelling,were monitored.Thereafter,24 successfullyestablished CIA rats were randomly assigned to 4 groups with 6 animals each:model,positive control drug,high-dose traditional Chinese herbal medicine,and traditional Chinese herbal medicine.A blank control group of 6 rats was included.After 12 weeks of intervention with Tengmei decoction,articular synovial tissue and serum specimens were collected to detect interleukin-2(IL-2)and IL-17 transcription and protein expression levels.

穿心莲内酯调控JAK2/STAT3信号通路对佐剂诱导的小鼠脚掌炎性肿胀的抗炎作用

目的:探讨穿心莲内酯对完全弗式佐剂(CFA)诱导的小鼠脚掌炎性肿胀的抗炎作用及其机制。方法:将60只昆明小鼠随机分为空白对照组、模型组、地塞米松(Dex)阳性对照组(2 mg/kg)以及穿心莲内酯高、中、低剂量组(300 mg/kg、150 mg/kg、75 mg/kg)。灌胃给药10 d后,除空白对照组外,其他各组均于小鼠右后足踝部注射CFA 20μL,复制小鼠足肿胀急性炎症模型。观察小鼠脚掌肿胀变化,计算脾脏指数,酶联免疫吸附试验(ELISA)法测定小鼠血清肿瘤坏死因子(TNF)-α水平;Western blot法检测磷酸化(p-)Janus激酶(JAK2)、p-信号转导子和转录激活子3(STAT3)蛋白表达。结果:与空白对照组比较,模型组小鼠脚掌明显肿胀,脾脏指数、TNF-α水平及p-JAK2、p-STAT3蛋白表达量明显升高(P<0.05);与模型组相比,穿心莲内酯可显著降低小鼠脚掌肿胀厚度以及脾脏指数,降低TNF-α的含量,抑制p-JAK2、p-STAT3的蛋白表达(P<0.05)。结论:穿心莲内酯可能通过阻断JAK2/STAT3信号通路,抑制CFA诱导小鼠脚掌炎性肿胀。

Analgesic and Anti-Arthritic Effect of <i>Enicostemma littorale</i>Blume

Enicostemma littorale (Blume), the folk medicine is used for rheumatic pain and it was selected for scientific validation. The 85% methanolic extract obtained from the whole plant of Enicostemma littorale has been assessed for analgesic and anti-inflammatory activity. The analgesic activity has been evaluated by hot plate and tail immersion method and the anti-inflammatory and antioxidant activities are evaluated by Complete Freund’s adjuvant induced arthritic model. The results of the evaluation of analgesic activity in hot plate and tail immersion method revealed that the extract exhibits significant activity at 150 mg/kg body weight and the effect is found to increase dose dependently. In Freund’s adjuvant induced arthritis, Enicostemma littorale is found to decrease the paw volume (15.81%). Significant protection is also observed by elevating antioxidant enzymes. In conclusion, the 85% methanolic extract of Enicostemma littorale possesses significant analgesic and anti-inflammatory activities in Freund’s adjuvant induced arthritic model in rats.

腊梅花精油的GC-MS分析及抗炎作用研究

目的明确十堰地区腊梅花精油的成分并在大鼠弗氏完全佐剂致足肿胀模型中探索其抗炎作用,为腊梅花资源的进一步开发提供依据。方法将SD大鼠随机分为空白对照组、模型组、阳性对照组(阿司匹林200 mg/kg)及腊梅花精油低、中、高剂量组(0.1、0.3、0.8 g/kg),灌胃给予相应药物或溶剂,给药容积10 mL/kg,1次/d。于第5天给药后30 min,大鼠右侧足跖皮下注射弗氏完全佐剂0.1 mL,空白对照组给予生理盐水0.1 mL。观察并用排水法测量致炎前后1、2、3、5、24、48、72 h足趾体积,计算各时间点足趾肿胀率,并用酶联免疫吸附试剂盒检测足部肿胀组织中炎症因子的水平。结果腊梅花精油中、高剂量组对弗氏完全佐剂致炎大鼠1、2、3、5、24、48 h的足趾肿胀均明显抑制(P<0.05),各剂量组肿胀足部组织中的TNF-α和IL-1β的含量明显减少。结论腊梅花精油对弗氏完全佐剂致大鼠足趾肿胀有明显的抑制作用,其抗炎作用的发挥可能与抑制TNF-α和IL-1β有关。

Anti-arthritic effect of Distemonanthus benthamianus extracts against rheumatoid arthritis in rats

Objective:To evaluate the anti-arthritic effect of aqueous and methanolic extracts of Distemonanthus benthamianus.Methods:Monoarthritis was induced by an injection of 0.3 mL zymosan A(0.9%NaCl,v/v)in the right posterior knee joints of rats.Then,joint diameter and pain threshold were determined.Polyarthritis was induced by an intracaudal injection of complete Freund’s adjuvant and rats were treated from day 14 post 1st complete Freund’s adjuvant injection until 28 day.The clinical,hematological,biochemical and oxidative stress parameters were evaluated.In addition,histological analysis of the knee joint was perfomed in both tests.Results:The aqueous and methanolic extracts of Distemonanthus benthamianus at a dose of 500 mg/kg ameliorated zymosan A-induced monoarthritis,as evidenced by reduced joint diameter,increased pain threshold,as well as improved joint architecture.In addition,both extracts of Distemonanthus benthamianus markedly increased body weight and pain threshold,while reducing paw edema in polyarthritic rats.They also led to a marked decrease in platelets and white blood cells(P<0.05),as well as a significant increase in red blood cells,hemoglobin and hematocrit(P<0.05).The aqueous and methanolic extracts of Distemonanthus benthamianus significantly reduced alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase activities,while increasing serum protein levels(P<0.05)with no significant variation in creatinine level.Moreover,both extracts increased catalase and glutathione activities(P<0.05),and inhibited malondialdehyde and nitric oxide production(P<0.01 and P<0.001)in the liver and kidneys.Histological analysis of the joints showed that both extracts triggered tissue reparation.Conclusions:Distemonanthus benthamianus could be used as a potential candidate in the treatment of rheumatoid arthritis.