银杏叶提取物增强大鼠脾脏和胸腺的免疫功能

目的研究银杏叶提取物(GBE)对大鼠脾脏和胸腺免疫功能的影响。方法给SD大鼠灌胃给予(40、120、360)mg/(kg·d)GBE,同时设置对照组。28 d后,水合氯醛麻醉处死大鼠,测量胸腺和脾脏的质量指数;MTT法检测伴刀豆球蛋白A(Con A)诱导的大鼠脾淋巴细胞增殖转化;中性红实验测定大鼠腹腔巨噬细胞吞噬功能;扫描电镜观察大鼠脾脏和胸腺的超微结构变化。结果不同剂量的GBE均可提高胸腺和脾脏的器官质量指数,不同剂量组之间无显著性差异;不同剂量的GBE均可增强大鼠腹腔内巨噬细胞的吞噬作用及各级淋巴细胞的增殖能力,且呈现一定的剂量依赖性。电镜观察发现不同剂量的GBE处理组,大鼠脾脏和胸腺中成熟淋巴细胞数均较对照组增多。结论银杏叶提取物可增强大鼠胸腺和脾脏的免疫功能。     

雌二醇对大鼠细胞免疫功能 的影响

本文采用大鼠去卵巢或再补充雌二醇(E_2)的方法探讨 E_(2)对免疫器官及其细胞免疫功能的影响。结果表明,去卵巢大鼠的脾脏和胸腺肥大,重量增加,脾细胞和胸腺细胞数明显增多。给去卵巢大鼠补充 E_2可使上述变化逆转。切除卵巢后的大鼠脾细胞对 ConA 诱导的增殖应答反应和诱生 IL—2的能力明显增高,补充 E_(2)亦可使上述变化逆转。但去卵巢大鼠的胸腺细胞对 ConA 诱导的增殖应答反应未能检测到可见的变化;给去卵巢鼠补充 E_2后,胸腺细胞对 ConA 的应答反应反而增强,并观察到胸腺细胞呈现轻度的自发增殖反应。实验结果表明:E_2能影响外周和中枢免疫器官及其细胞免疫功能,但这种影响在外周和中枢免疫器官表现出不同的变化。     

脾脏切除对胸腺和淋巴结免疫功能的影响

报道脾脏切除（脾切）对动物胸腺和淋巴结免疫功能的影响，脾切后３ｗ大鼠胸腺细胞自发增殖及ＣｏｎＡ诱导增殖均无明显变化，ＣｏｎＡ诱导淋巴结细胞增殖脾切组明显高于假手术对照组，脾切后３ｗ再用ＳＲＢＣ免疫，５ｄ后小鼠腹腔淋巴结数量脾切明显高于对照组，用ＱＨＳ法测定淋巴结细胞对ＳＲＢＣ初次抗体产生脾切组则明显低于对照组。     

不同生长期小鼠胸腺T细胞不同亚群的变化及ROCK抑制剂对衰老小鼠胸腺再生的促进作用

目的研究各发育阶段小鼠胸腺细胞、胸腺上皮细胞(TEC)和脾脏中T细胞的变化,探索Rho相关卷曲蛋白激酶(ROCK)抑制剂对衰老小鼠胸腺再生的作用。方法取幼年、青年、中年及老年期C57BL/6雌鼠的胸腺和脾脏,流式细胞术分析小鼠胸腺细胞、TEC及脾脏中T细胞亚群;ROCK抑制剂体外处理衰老进程中小鼠TEC,荧光酶联斑点分析仪观察细胞增殖情况;ROCK抑制剂处理20月龄衰老小鼠,流式细胞术检测小鼠胸腺细胞、TEC及脾脏中T细胞亚群的变化。结果随月龄增加,胸腺细胞、TEC总数及各细胞亚群数量均显著降低;脾脏中总CD4⁺T细胞及CD8⁺T细胞比例无显著性改变,但CD4+初始T细胞、CD8+初始T细胞和CD4+近期胸腺迁出细胞(RTE)在脾脏中所占比例呈下降趋势;ROCK抑制剂在体外促进衰老进程中小鼠TEC增殖,ROCK抑制剂在体内使衰老小鼠胸腺细胞、TEC以及外周脾脏中T淋巴细胞各亚群数量明显增加。结论随着小鼠月龄增加,胸腺呈进行性退化。ROCK抑制剂可以显著缓解衰老小鼠胸腺的萎缩,促进衰老小鼠胸腺再生。     

L783白血病小鼠体内的免疫活性细胞和免疫球蛋白的变化

本文报道沪白 1号(SW1)近交系小鼠腹腔接种L783白血病小鼠脾脏细胞悬液后,胸腺、脾脏和淋巴结中T和B淋巴细胞百分率以及血清中 IgG和 IgM含量的变化。结果表明,淋巴器官除胸腺外,脾脏和淋巴结中的T淋巴细胞百分率明显增高,而B淋巴细胞显著下降。血清中Ig 含量未见明显变化。上述免疫活性细胞有规律性的变化为进一步研究白血病发病机理提供线索,并有可能作为考核抗白血病药物疗效的指标之一。     

T细胞定向迁移诱导实验性自身免疫性脑脊髓炎小鼠胸腺萎缩

目的 初步探讨实验性自身免疫性脑脊髓炎(EAE)小鼠胸腺萎缩的机制.方法 髓鞘少突胶质细胞糖蛋白(MOG)免疫C57BL/6小鼠诱导EAE,卵清白蛋白(OVA)免疫的小鼠作为对照;不同时间点计数胸腺、脾脏、淋巴结细胞总数,检测脾脏中胸腺来源细胞及中枢神经系统(CNS)浸润细胞.结果 MOG肽成功诱导EAE动物模型,小鼠出现典型的肢体运动功能障碍,脊髓可见大量炎性细胞浸润;MOG和OVA免疫均诱导胸腺细胞增加,第5天达到高峰,随后逐渐下降;EAE发病后胸腺细胞迅速减少,发病高峰期几乎完全消失,胸腺严重萎缩;MOG和OVA免疫后脾脏和淋巴结细胞总数持续升高,新近胸腺来源的T细胞增加尤其明显;EAE发病后脾脏T细胞总数减少,CNS浸润淋巴细胞总数增加.结论 大量T细胞在胸腺发育成熟并释放到外周,进而定向迁移至CNS诱导EAE是胸腺萎缩的主要原因.     

过氧化物酶体增殖剂激活受体α对小鼠T、B细胞发育的影响

目的研究过氧化物酶体增殖剂激活受体α(PPARα)与小鼠免疫系统功能和发育的关系。方法喂养含过氧化物酶体增殖剂(PP)的食物后,观察野生型C57Bl/6小鼠和PPARα缺陷型小鼠胸腺、脾脏的重量及胸腺细胞、脾细胞数量的变化。用抗小鼠CD3、CD4、CD8a、CD19、IgM或CD45R/220单克隆抗体(mAb)进行细胞免疫荧光染色后,用流式细胞术分析骨髓细胞、胸腺细胞和脾细胞的表型变化。用刀豆蛋白A(ConA)和脂多糖(LPS)分别刺激小鼠的脾细胞,用3H-TdR掺入法检测淋巴细胞增殖活性的变化。用RT-PCR检测小鼠骨髓、胸腺和脾脏中PPARαmRNA的表达。结果与野生型小鼠相比较,PP对PPARα缺陷型小鼠胸腺、脾脏的重量和细胞数,以及ConA和LPS刺激对淋巴细胞增殖反应的影响很小。PP可致野生型小鼠胸腺中CD4 CD8 T细胞数,骨髓中B220 B细胞总数和原/前B细胞数明显减少,但对PPARα缺陷型小鼠的上述T、B细胞亚群无显著影响。PPARαmRNA在小鼠胸腺和脾脏中呈低表达,在骨髓中不表达。结论PPARα在PP诱导的免疫调节中起主要作用,其可通过间接途径影响T、B细胞的发育。     

胸腺细胞凋亡在EAE模型中的变化

目的研究EAE大鼠胸腺细胞凋亡的变化规律,为探讨胸腺在EAE发病中的作用提供理论依据。方法应用光镜、电镜观察正常组、EAE发病第3天组、恢复期组大鼠胸腺细胞变化情况,应用caspase-3免疫组化染色及流式细胞术观察比较胸腺细胞凋亡情况。结果电镜、光镜观察显示正常组和恢复期组胸腺结构基本一致,皮质髓质分界清楚,淋巴细胞密集,核呈蓝紫色,发病第3天组胸腺明显萎缩,皮质变薄,淋巴细胞数减少,并可见凋亡淋巴细胞;caspase-3免疫组化染色显示发病第3天组胸腺皮质凋亡淋巴细胞数目明显增多;流式细胞术显示发病第3天组大鼠胸腺细胞凋亡率显著升高(P<0.05),恢复期组凋亡率与正常组相比无统计学意义(P>0.05)。结论 EAE大鼠发病后第3天胸腺细胞明显减少并发生凋亡,随病情恢复淋巴细胞增加、凋亡细胞减少,表明胸腺具有自我恢复能力,其变化与EAE形成有关。     

HBV转基因小鼠肝脏NKT细胞功能与PD1、CD28表达分析

目的研究HBV转基因小鼠肝脏中NKT细胞的功能与表面PD1、CD28表达的关系。方法分离小鼠肝脏、脾脏、胸腺和腹膜淋巴结单个核细胞,利用流式细胞检测技术,分别检测其淋巴细胞中NKT细胞的频率,同时检测肝脏NKT细胞PD1、CD28的表达及IFN-γ、IL-4的分泌功能,比较肝脏、脾脏、胸腺和腹膜淋巴结这几个主要免疫组织淋巴细胞中NKT细胞所占的比例,并分析肝脏NKT细胞PD1、CD28的表达与细胞功能的关系。结果与正常同品系小鼠比较,HBV转基因小鼠肝脏、脾脏、胸腺和腹膜淋巴结NKT细胞数量明显减少(P<0.05),与脾脏、胸腺和腹膜淋巴结相比,肝脏淋巴细胞中含有大量的NKT细胞;与正常同品系小鼠比较,HBV转基因小鼠肝脏NKT细胞PD1的表达明显增多(P<0.05),CD28的表达明显减少(P<0.05),肝脏NKT细胞IFN-γ、IL-4的分泌功能明显降低(P<0.05)。结论肝脏中含有大量的NKT细胞,HBV转基因小鼠肝脏NKT细胞的功能存在明显的缺陷,并提示PD1的增加和CD28的降低可能与NKT细胞功能的下调密切相关。     

摄入不同剂量过量碘大鼠脾脏的形态学改变

目的对摄入不同剂量过量碘的大鼠脾脏进行形态学研究,探讨碘过量对大鼠机体免疫功能的影响。方法断乳1个月Wistar大鼠18只,雌雄各半,分为适碘组(NI)、10倍碘组(10HI组)和100倍碘组(100HI组)。给以不同浓度碘化钾水喂养3个月后,处死取脾脏并称重,对脾脏进行光镜下观察,并对脾小结、生发中心、动脉周鞘和边缘区进行体视学定量分析,对脾脏边缘区免疫细胞进行超微结构观察。结果与正常组比较,10HI组和100HI组大鼠脾脏绝对重量和脾脏指数未见明显改变,10HI组大鼠脾脏脾小结、生发中心、动脉周鞘和边缘区体密度未见明显改变,脾脏免疫细胞超微结构未见明显改变。100HI组大鼠脾脏脾小结、生发中心、动脉周鞘和边缘区体密度明显增加,脾脏免疫细胞超微结构出现明显改变,即呈现功能活跃的表现。结论低剂量过量碘摄入对大鼠免疫功能未产生明显影响,高剂量过量碘摄入可引起大鼠免疫功能亢进。     

加味四逆散对慢性心理应激大鼠胸腺糖皮质激素受体作用的研究

目的:观察加味四逆散对慢性心理应激大鼠胸腺细胞糖皮质激素受体作用的影响。方法:大鼠随机分为正常对照组(C组)、模型组(M组)、加味四逆散组(C₁组)、人参皂甙组(C₂组)。采用放射免疫方法检测大鼠胸腺细胞糖皮质激素受体的数目、核转移率。结果:M组胸腺重量指数显著低于C组,糖皮质激素受体(GCR)的数目差异不显著;胸腺糖皮质激素受体的核转移率显著高于C组(P<0.01)。C₁组与C₂组的大鼠胸腺细胞糖皮质激素受体复合物的核转移率均显著低于M组,胸腺重量指数显著高于M组(P<0.01或P<0.05)。在受体核转移率下降方面,C₁组作用大于C₂组(P<0.05)。此外,C₁组各项检测数据与C组相比无统计学差异。结论:加味四逆散能显著减轻糖皮质激素对胸腺的抑制作用。其作用途径可能是通过抑制慢性心理应激大鼠GCR由胸腺细胞胞浆向胞核转位来实现的。     

淫羊藿多糖致小鼠胸腺缩小的免疫药理机理研究

淫羊藿多糖(EPS)刺激外周T 细胞功能的同时,引起胸腺缩小。我们发现EFS10～50mg/kg 范围内.胸腺缩小伴随着胸腺内L3T4~+和Lyt2~+细胞数减少,对有丝分裂原的刺激反应降低;但此时胸腺细胞增殖和产生IL-2能力提高。单次大剂量(100mg/kg)皮下注射.能够引起小鼠胸腺细胞释放增加,P<0.01。由此我们认为,EPS 促进胸腺释放成熟细胞是其导致胸腺缩小的机理之一,也是其增强机体细胞免疫功能的重要机制。     

Hedgehog信号通路阻断剂环巴胺体外对佐剂性关节炎大鼠的关节软骨细胞增殖的影响

目的探讨Hedgehog(Hh)信号通路抑制剂环巴胺体外对佐剂性关节炎大鼠(AA)模型的关节软骨细胞增殖的影响和部分机制。方法弗氏完全佐剂诱导AA大鼠,测量关节炎指数和继发性足肿胀度,HE染色观察两组软骨组织生长情况;取AA大鼠踝关节软骨组织,采用胰蛋白酶-胶原酶法分离、培养、鉴定,环巴胺(0、0.05、0.5、5、20μmol/L)体外给药,MTT法检测AA大鼠踝关节软骨细胞增殖,Annexin V-FITC/PI双染检测AA大鼠踝关节软骨细胞凋亡,Western blotting检测AA大鼠踝关节软骨细胞Shh、Ptch1、Gli1的蛋白表达。结果弗氏完全佐剂诱导后,与正常大鼠相比,AA大鼠关节炎指数和继发性足肿胀度明显升高,HE染色显示,AA大鼠踝关节软骨组织有破坏;甲苯胺蓝和Ⅱ型胶原鉴定体外成功培养AA大鼠踝关节软骨细胞;体外给药环巴胺(0.05、0.5、5、20μmol/L)可升高AA模型关节软骨细胞增殖,流式细胞检测结果显示,环巴胺能降低AA模型软骨细胞凋亡率;与未用环巴胺组相比,环巴胺(0.5、5、20μmol/L)给药对AA软骨细胞中Hh信号通路相关蛋白(Shh、Ptch1、Gli1)表达显著下降。结论弗氏完全佐剂诱导建立AA大鼠模型成立,体外给药环巴胺可抑制AA大鼠软骨细胞的增殖,抑制软骨细胞的凋亡,该作用与抑制AA大鼠软骨细胞Hh信号有关。     

Effects of exogenous cytokines on the ethanol-mediated suppression of murine thymocyte proliferation

Although attempts have been made to assess the effect of ethanol on murine thymocyte proliferation, the mechanism which accounts for the immunosuppressive effect of ethanol on the thymocyte proliferation has not been elucidated. Thus, a mouse model was used to determine (1) whether there is a similarity in the effect of ethanol exposure in vitro and in vivo on the proliferative response of thymocytes to phytohemagglutinin (PHA), (2) whether ethanol exposure affects the responsiveness of thymocytes to exogenous interleukin (IL)-I and IL-2, and (3) whether ethanol affects IL-1 production by peritoneal macrophages. We found that the proliferative response of thymocytes from mice fed on an ethanol-containing diet was significantly inhibited (P<0.05) compared to that in mice red on maltose or standard diets. We also observed that low concentrations of ethanol (12.5 mM) appeared to enhance the mitogenic response of thymocytes to PHA, but the response was not significantly greater than that of controls (P>0.05). Ethanol at higher concentrations (25–100 mM) significantly suppressed the mitogenic response of thymocytes to PHA (P<0.05) in a dose-dependent manner. Our data also revealed that (1) ethanol did not significantly suppress IL-I secretion by adherent macrophages stimulated by LPS, and (2) the addition of exogenous IL-I was insufficient to restore full responsiveness in thymocytes from ethanol-fed mice. Taken together, these results suggest that the suppressive effect of ethanol on thymocyte proliferation is not mediated by insufficient IL-1. Finally, we present novel evidence that addition of exogenous IL-2 completely restores the impaired proliferative response of thymocytes from ethanol-fed mice to control levels. In summary, our results demonstrate that ethanol inhibits thymocyte proliferation in response to PHA, and that the inhibition is not due to insufficient IL-1. We also report that addition of exogenous IL-2 is sufficient to restore full proliferative capacity to thymocytes from ethanol-fed mice.     

Modulating effect of mast cells on the proliferation of cells from various lymphoid organs of rats]

There are some reasons for suggesting that mast cells (MC) can modulate lymph cell (LC) activity. The blast-transformation test was used to study the effect of serum MC of Wistar and ACI rats on the proliferation of splenic, thymus and lymph node LC in vitro. Rat MC modulate lymphocyte proliferation. Depending on their concentrations, serum MC increased spontaneous and T-mitogen induced proliferation of splenic and lymph node cells. T-mitogen thymocyte proliferation was also increased. The promoting effect was maximal when the MC proliferative response was induced by suboptimal doses of mitogens. No genetical restriction was found for development of the MC promoting effect.     

板蓝根凝集素对小鼠胸腺发育的影响

目的：实验研究板蓝根凝集素对小鼠胸腺细胞发育和胸腺细胞增殖的作用。方法：用生物化方法提取板蓝根凝集素,以昆明种小鼠为观察对象,用０．０５４ｍｇ／ｋｇ／ｄ剂量板蓝根凝集素灌胃,连续３０天,采用组织切片方法,江学显微镜检测胸腺皮质与髓质细胞,并进行生物学统计。结果：与对照组比较,实验组小鼠胸／体比值Ｐ＜０．０５,胸腺皮质与髓质胸腺细胞密度Ｐ＜０．０１,细胞直径＞０．０５。结论：实验证明板蓝根凝集素对小鼠胸腺的发育和胸腺细胞的增殖可能具有促进作用,对维持胸腺微环境可能起间接作用。     

通痹灵总生物碱抑制关节破坏的细胞免疫作用机制

目的:证实通痹灵总生物碱抑制胶原诱导型关节炎(CIA)大鼠关节炎症和关节破坏的药效,揭示其调节细胞免疫的作用机制。方法:牛II型胶原混合弗氏不完全佐剂于SPF级Wistar大鼠尾根部注射,足肿后随机分为造模加生理盐水组,甲氨蝶呤(MTX)治疗组和通痹灵总生物碱(以马钱子碱和士的宁为主的生物碱,TBL总碱)治疗组,容积法评价后肢肿胀度;给药后35d,乳腺钼靶机大鼠全身摄片,计分法评价各组大鼠X光片四肢96块骨侵蚀和100个关节间隙变化;处死动物,取左前肢和右后肢近端第3足趾关节苏木素-伊红(HE)染色,评价中性粒、淋巴细胞、浆细胞浸润、滑膜细胞增生的情况;MTT法检测TBL总碱对Jurkat细胞增殖的影响;Western blot检测TBL总碱对Jurkat细胞ERK1/2磷酸化的影响。结果:造模成功后,MTX和TBL总碱给药35d都可明显抑制大鼠关节肿胀和关节破坏,与造模加生理盐水组有统计学差异(P<0.01);TBL总碱可明显抑制CIA淋巴细胞、浆细胞的浸润和滑膜增生(P<0.05),MTX可抑制滑膜增生。TBL总碱可显著抑制T细胞增殖以及ERK1/2的磷酸化。结论:TBL总碱具有抑制关节免疫炎症和关节破坏的作用,其机制可能是通过干预T细胞MAPK信号转导,抑制T细胞的活化和增殖来实现的,为TBL总碱治疗RA提供了实验依据。     

Endocrine Pancreas Histology of Congenic BB-Rat Strains with Reduced Diabetes Incidence After Genetic Manipulation on Chromosomes 4, 6 and X

Congenic BB.SHR rat strains were established by crossing of spontaneously diabetic BB/OK rats and diabetes-resistant SHR rats. Chromosomal regions on which the genes Iddm 4 (BB.6s), Iddm6 (BB.Xs) and Iddm 2 (BB.LL) are located were exchanged. As a result of genetic manipulation diabetes incidence was markedly reduced from 80% in BB/OK to 50% in BB.SHR (Chr. X), to 14% in BB.SHR (Chr. 6) and to 0% in BB.LL rats. Pancreata of these newly generated BB.SHR rats were investigated histologically. In newly diagnosed diabetic rats of congenic strains pancreatic insulin content (BB.6s: p<0.05; BB.Xs p<0.01) and relative volume of insulin-positive cells (BB.Xs: p<0.001) were significantly higher than in BB/OK rats. The degree of insulitis was not different in 90-day-old and newly diagnosed diabetic animals. Surprisingly, in 30-day-old rats we observed an increase of the degree of insulitis with decreasing diabetes incidence. We suppose that by an earlier occurrence of the immunological β-cell destruction, a part of the animals is able to develop a secondary diabetes resistance. The exchange of the BB-lymphopenia gene by that of SHR-rats prevented the development of hyperglycaemia without altering the auto-reactive immune response, which could be observed in all animals investigated.     

Anti-inflammatory effects of serum isolated from animals on intermittent feeding in C6 glioma cell line

Multiple sclerosis (MS) is a demyelinating disease of the CNS. Early inflammation leads to later destruction of myelin in MS. Dietary restriction (DR) produces anti-inflammatory and immunomodulatory effects in many species. Based on the reported anti-inflammatory effects of DR, we investigated whether sera collected from rats fed on intermittent feeding (IF, a type of DR) diet could modulate cytokine secretion and matrix metalloproteinase (MMP-2) activity that are involved in MS pathogenesis. Cytokine levels (IL-6 and TGF-β1) were measured in supernatant from C6 glioma cell line cultures treated with IF and AL fed animals' sera by enzyme-linked immunosorbent assay (ELISA) and MMP-2 activity was detected by gelatin zymography. Our results indicated that sera of animals on IF diet significantly reduced IL-6 (p<0.05) and increased TGF-β1 (p<0.05) production by C6 glioma cells. A significant decrease (p<0.05) in MMP-2 activity was also found. These results indicate anti-inflammatory and immunomodulatory activity in the sera of animals on IF regimen on cells involved in multiple sclerosis pathogenesis. Further studies on the detection of factors responsible for such activities and their mechanism of action in MS pathogenesis are recommended.