饵料维生素E含量对酸应激中华鳖幼鳖血清补体C3和C4的影响

为探讨维生素E(VE)对中华鳖(Pelodiscus sinensis)幼鳖血清补体C3和C4的影响及补体在酸应激下的变化,在6组(对照组,实验Ⅰ、Ⅱ、Ⅲ、Ⅳ和Ⅴ组)幼鳖的饵料中依次添加0、50、250、500、1000和5000mg／kg的VE,喂食4周,每组取半数幼鳖经酸应激处理24h。取幼鳖血清,用透射比浊法测定血清补体C3和C4的含量。经和未经酸应激的实验Ⅱ、Ⅲ和Ⅳ组幼鳖血清补体C3的含量均明显高于对照组,实验Ⅰ和Ⅱ组C4含量也明显高于对照组;经酸应激的幼鳖与未经酸应激的比较,对照组和实验Ⅰ组C3和C4的含量显著下降,其余4组没有变化。分析说明,VE在一定剂量范围内能促进血清补体C3和C4的合成,酸应激能导致其下降;而高剂量的VE对酸应激导致的不利影响有拮抗作用。     

维生素C和酸应激对中华鳖幼鳖血清补体C3和C4含量的影响

为研究维生素C对中华鳖(Pelodiscus sinensis)血清补体C3和C4的影响及其在酸应激条件下的变化,我们设置了6个实验组,饵料中维生素C的添加量依次为0、250、500、2500、5000和10000mg／kg,喂食4周后取其血清,用透射比浊法测定酸应激前后中华鳖血清补体C3和C4的含量。结果表明,维生素C添加量为250mg／kg时,血清补体C3的含量与对照组间没有明显不同;维生素C添加量为500、2500、5000和10000mg／kg的4组,血清补体C3的含量明显高于对照组和维生素C添加量为250mg／kg组;维生素C添加量为500mg／kg的一组,血清补体CA含量明显高于其它5组;维生素C添加量为250mg／kg组明显高于10000mg／kg组。酸应激后,补体C3的含量没有明显下降,将维生素C添加量为0、250和500mg／kg的三组并为一组处理,则应激后有明显下降。维生素C添加量为0、250和500mg／kg的3组,血清补体CA的含量在酸应激后明显下降,而维生素C添加量为2500、5000和10000mg／kg的3组,应激后血清补体C4没有明显变化。维生素C和酸应激对中华鳖血清补体C3和CA含量的影响没有交互作用。这说明,维生素C在一定剂量范围内,能提高中华鳖血清补体C3和CA的水平,酸应激能导致其含量降低,而高剂量的维生素C对其下降有颉颃作用[动物学报49(6)：769～774,2003]。     

维生素C和E混合饲喂对中华鳖幼鳖抗酸应激能力的影响

来自甲鱼养殖场的60只中华鳖(Pelodiscus sinensis)幼鳖驯养3周后,实验设5组：对照组、处理Ⅰ、Ⅱ、Ⅲ和Ⅳ组。各组设2个平行,依次在饵料中混合添加维生素C(Vc)和E(VE)为0和0、250和50、2500和50、250和250、2500和250mg／kg,喂食4周后,每组取半数幼鳖经酸应激处理24h。取幼鳖血液,用镜检法测定血细胞的吞噬率,透射比浊法测定血清溶菌活力、杀菌活力以及补体C3和C4含量。①经酸应激与未经酸应激处理相比：对照组血细胞吞噬率显著降低,而处理Ⅰ-Ⅳ组无显著变化;对照组和处理Ⅰ组血清溶菌活力和补体C3含量显著下降,而处理Ⅱ-Ⅳ组无显著变化;血清杀菌活力均有显著下降(对照组、处理Ⅰ和Ⅲ组极显著,处理Ⅱ组和Ⅳ组显著);对照组、处理Ⅰ和Ⅲ组血清补体C4显著下降,而处理Ⅱ和Ⅴ组无显著变化。②经酸应激处理,血细胞吞噬率、血清溶菌活力、杀菌活力和补体C3含量,处理Ⅰ～Ⅳ组的均显著高于对照组,处理Ⅳ组显著高于其他4组;血清杀菌活力,处理Ⅱ组又高于处理Ⅰ和Ⅲ组;血清补体C4,对照组显著低于处理Ⅰ-Ⅳ组,而处理Ⅰ-Ⅳ组间无显著相异。Vc和VE混合饲喂对酸应激后中华鳖血细胞吞噬率、血清溶菌活力、杀菌活力和补体C3含量有显著协同促进作用,对血清补体C4的合成无协同作用。说明Vc和VE混合饲喂能显著增强中华鳖抗酸应激能力,缓解或部分缓解酸应激造成的不利影响。     

维生素C和E合用对应激和非应激中华鳖幼鳖生长、肝脏维生素C和E以及血清皮质醇含量的影响

为探讨维生素C (VC)和维生素E (VE)联用对应激和非应激中华鳖幼鳖的生长、肝脏VC 和VE 以及血清皮质醇含量的影响 ,作者使用了 5组饵料 ,VC 和VE 的添加量依次为 0和 0mg/kg (对照组 )、 2 5 0和 5 0mg/kg (实验Ⅰ组 )、2 5 0 0和 5 0mg/kg (实验Ⅱ组 ) ;2 5 0和 2 5 0mg/kg (实验Ⅲ组 ) ;2 5 0 0和 2 5 0mg/kg(实验Ⅳ组 )。中华鳖幼鳖的生长、肝脏VC 和VE以及血清皮质醇分别通过特定生长率、高压液相色谱法和放免法来测定。结果实验Ⅰ -Ⅳ组中华鳖的特定生长率明显高于不加VC 和VE的对照组 ,但实验Ⅰ -Ⅳ组间没有明显不同。非应激中华鳖肝脏VC 和VE的含量随饵料中VC 和VE 含量的增加而明显升高 ,并且实验Ⅱ -Ⅳ组肝脏VC 和VE都明显高于对照组和实验Ⅰ组。酸应激后 ,对照组和实验组中华鳖肝脏VC 和VE都有下降的趋势 ,但无显著差异 ;应激后实验Ⅱ -Ⅳ组肝脏VC 和VE均明显高于对照组 ,实验Ⅳ组明显高于其它 4组。血清皮质醇的含量在实验Ⅰ -Ⅳ组间没有明显不同 ,实验Ⅰ -Ⅲ组与对照组相比虽有降低的趋势但没有变化 ,实验Ⅳ组则明显低于对照组。酸应激后 ,对照组血清皮质醇明显升高 ,其他 4组虽有升高的趋势 ,但没有明显变化。应激后实验Ⅰ -Ⅳ组血清皮质醇的含量均明显低于对照组 ,实验Ⅰ -Ⅳ组间没有明显     

维生素C和E合用对中华鳖幼鳖非特异性免疫功能的影响

本研究旨在探讨维生素C和E合用对中华鳖幼鳖非特异性免疫功能的影响。实验设 5组 ,5组饵料中维生素C和E的添加量依次为 0和 0mg/kg(对照组 ) ;2 5 0和 5 0mg/kg(实验Ⅰ组 ) ;2 5 0 0和 2 5 0mg/kg(实验Ⅱ组 ) ;2 5 0和2 5 0mg/kg(实验Ⅲ组 ) ;2 5 0 0和 2 5 0mg/kg(实验Ⅳ组 )。喂食 4周 ,取其血液 ,测定中华鳖血细胞的吞噬率、血清溶菌活力、杀菌活力、补体C3和C4。结果表明 ,实验Ⅱ—Ⅳ组 ,血细胞吞噬率和血清杀菌活力明显高于对照组 ,实验Ⅳ组明显高于其他 4组 ,并且维生素C和E之间有明显的协同作用。血清溶菌活力和补体C3的含量 ,在实验Ⅰ—Ⅳ组明显高于对照组 ,实验Ⅳ组明显高于其他 4组。补体C4在实验Ⅰ—Ⅳ组明显高于对照组 ,但实验Ⅰ—Ⅳ组间没有明显差别 ,维生素C和E间也没有协同作用。     

外源维生素C及冬眠对中华鳖稚鳖生长、维生素C合成和肝脏维生素C含量的影响

本文研究了饲料中维生素C(Vc)对中华鳖稚鳖冬眠前后Vc合成能力及肝脏中Vc含量的影响。将体重为29.41g±5.44g的稚鳖180只,按其饲料中Vc含量(0、500、2500mgVc/kg饲料)分为Vc0、Vc500和Vc2500共3组,每组6个平行,每平行10只鳖。实验开始前称重得各组初始平均体重。25℃±1℃下用实验饲料饲喂4周后,缓慢降温至7℃,人工诱导其进入冬眠,此时为冬眠期开始,每组取6只鳖,称重后快速处死,取其肝脏和肾脏。其余鳖全部称重。冬眠8周末取样,处理同上。用高效液相色谱(HPLC)法测定肝脏Vc含量和肝脏、肾脏古洛糖酸内酯氧化酶(GLO)活力。结果发现中华鳖自身能够合成Vc,且合成部位在肾脏。饲喂4周不同Vc水平的饲料后,各组间特定生长率差异不显著。冬眠8周后,各组体重均有所降低,组间冬眠期间特定生长率无显著性差异。冬眠开始时,稚鳖肝脏Vc蓄积量有随外源Vc浓度升高而递增的趋势,但差异未达到显著性水平;经历冬眠后各组稚鳖肝脏Vc含量降低,Vc500组下降不显著,而Vc0与Vc2500组显著下降;肾脏GLO活力在不同Vc梯度组间及冬眠前后均无显著差异。以上结果表明:中华鳖冬眠过程中要消耗较多的肝脏Vc,外源Vc和冬眠对肾脏Vc合成能力影响不显著。     

慢性氨暴露对中华鳖幼鳖生长及非特异性免疫反应的影响

在高密度养殖模式下,中华鳖经常暴露于高氨环境中。本研究评价了慢性氨暴露对中华鳖(Pelodiscussinensis)幼鳖生长及非特异性免疫功能的影响。将鳖(体重90.5±20.5g)饲养于非离子氨浓度分别为1.49mg/L(C1)、2.61mg/L(C2)和4.14mg/L(C3)的环境中84d,以不外加氨氮的自来水饲养组为对照(C0),实验期间各处理组均保持恒定的温度(29.5±0.5℃)和pH(7.8±0.1)值。氨暴露21、42和84d后测体重,氨暴露84d后取各实验组鳖的血样。在本实验设定浓度范围内,氨对中华鳖的生长及非特异性免疫指标如血清溶血活性、血清杀菌活性、血清溶菌活性和脾脏系数没有显著影响。但氨暴露组中华鳖血液和脾脏淋巴细胞α-乙酸萘酯酶(ANAE)阳性率与对照组相比均显著下降。研究结果表明中华鳖对高氨耐受能力比鱼类更强。     

过敏性紫癜合并肺炎支原体感染患儿呼吸道菌群多样性与补体C3、C4及免疫指标的相关性

目的研究儿童过敏性紫癜(HSP)合并肺炎支原体(MP)感染患儿呼吸道菌群多样性与补体C3、C4及免疫指标的相关性。方法选择2017年3月至2020年3月于我院接受治疗的HSP患儿96例,其中48例合并MP感染的患儿设为合并MP组,48例未合并MP感染的患儿设为HSP组。另外选择同期我院健康体检儿童45例为对照组。比较3组对象呼吸道菌群多样性,血清免疫球蛋白IgG、IgA、IgM、IgE及补体C3、C4水平,并分析呼吸道菌群多样性与免疫指标间的相关性。结果合并MP组患儿呼吸道菌群Simpson指数显著高于对照组,Shannon指数显著小于对照组(均P<0.05)。HSP组患儿呼吸道菌群Simpson指数及Shannon指数与对照比较差异无统计学意义(均P>0.05)。合并MP组患儿血清IgA水平较对照组显著上升,IgM水平显著下降;HSP组患儿血清IgA、IgE水平较对照组显著上升;合并MP组患儿血清IgE、IgM水平显著低于HSP组(均P<0.05);而3组对象血清IgG水平比较差异无统计学意义(均P>0.05)。合并MP组和HSP组患儿血清补体C3水平显著低于对照组;合并MP组患儿血清补体C4水平显著低于对照组;合并MP组患儿血清补体C3、C4水平均显著低于HSP组(均P<0.05)。IgM、IgE及补体、C3、C4水平与Simpson指数呈负相关,与Shannon指数呈正相关(均P<0.05)。结论HSP合并MP感染患儿呼吸道菌群多样性与补体C3、C4及IgM、IgE水平具有显著相关性。     

流行性出血热患者血清补体C3.C4的检测及其动态观察

对96例EHF患者血清补体C3、c4的检测及动态观察结果表明:EHF患者补体C3、C4均值都高于正常人,其中以C4增高较明显。根据EHF患者血清中补体含量的不同,可分为C3水平低下组,C3或C4水平增高组,C3或C4水平正常组。EHF患者补体水平的变化是极为复杂的,既存在有补体水平的下降,又存在有补体水平的升高。本文对此研究结果进行了讨论。     

补体C3对大肠杆菌疫苗的免疫增强作用

从SPF鸡血清中分离纯化C3,用戊二醛将其与大肠杆菌抗原连接,免疫注射SPF鸡;对照组鸡免疫注射弗氏完全佐剂大肠杆菌疫苗。分别在免疫后的第2、3、4、5、6、7、8、9周采血,用ELISA方法测定血清中的抗体含量,同时测定血清中总补体活性。结果表明,免疫后3周,弗氏佐剂大肠杆菌疫苗诱导鸡体产生的抗体效价高于C3佐剂疫苗组,但至第4周,弗氏佐剂疫苗组的抗体水平达到高峰(OD值=0.270±0.004),然后迅速下降,到第9周降至0.200±0.005,而C3疫苗组鸡免疫后的抗体水平持续上升,从免疫后第2周的0.098±0.003上升到第8周的0.275±0.002。证明C3能够促进免疫记忆细胞的产生,并能够使细菌抗原给予免疫细胞持续稳定的刺激,从而使鸡体维持高水平抗体的时间延长。研究结果为研制有效的家禽细菌性疫苗提供了理论依据。     

The effects of dietary vitamin C on growth,liver vitamin C and serum cortisol in stressed and unstressed juvenile soft-shelled turtles (Pelodiscus sinensis)

We studied the effect of dietary vitamin C on growth, liver vitamin C and serum cortisol levels in stressed and unstressed juvenile soft-shelled turtles. Turtles were fed with vitamin C supplementation at dosages of 0, 250, 500, 2500, 5,000 or 10,000 mg/kg diet for 4 weeks. Vitamin C supplementation exerted significant effects on specific growth rate and liver vitamin C concentrations. The specific growth rate peaked in the group fed at 500 mg/kg diet, while liver vitamin C levels increased with increasing dietary vitamin C levels. Serum cortisol concentrations did not differ between groups of turtles fed diets supplemented with vitamin C in the range of 0-10,000 mg/kg. After acid stress, hepatic vitamin C levels were unaffected, while serum cortisol in the control group was significantly elevated (P<0.01). The other five groups of turtles did not show significant changes in serum cortisol compared with pre-stress levels.     

Complement C3 opsonization of Chlamydia trachomatis facilitates uptake in human monocytes

Chlamydia trachomatis is an obligate intracellular bacterium that causes severe infections, which can lead to infertility and ectopic pregnancy. Although both innate and adaptive immune responses are elicited during chlamydial infection the bacterium succeeds to evade host defense mechanisms establishing chronic infections. Thus, studying the host–pathogen interaction during chlamydial infection is of importance to understand how C. trachomatis can cause chronic infections. Both the complement system and monocytes play essential roles in anti-bacterial defense, and, therefore, we investigated the interaction between the complement system and the human pathogens C. trachomatis D and L2.Complement competent serum facilitated rapid uptake of both chlamydial serovars into monocytes. Using immunoelectron microscopy, we showed that products of complement C3 were loosely deposited on the bacterial surface in complement competent serum and further characterization demonstrated that the deposited C3 product was the opsonin iC3b. Using C3-depleted serum we confirmed that complement C3 facilitates rapid uptake of chlamydiae into monocytes in complement competent serum. Complement facilitated uptake did not influence intracellular survival of C. trachomatis or C. trachomatis-induced cytokine secretion. Hence, C. trachomatis D and L2 activate the complement system leading to chlamydial opsonization by iC3b and subsequent phagocytosis, activation and bacterial elimination by human monocytes.     

Measurement by ELISA of Complement Factor 4 (C4) in the Rat Brain: Necessity for Removal of Cerebrovascular Proteins

Assessment of complement 4 (C4) levels in experimental animals is used as a marker for activation of the classical complement pathway. The objective of this study was to develop a method for measuring C4 concentrations in the rat brain. An ELISA (sensitivity = 0.5 ng C4/ml) was used to measure C4 in regional brain homogenates from Fischer rats cardiac-perfused with phosphate buffered saline to remove cerebrovascular contents, and from sham-perfused rats. Ventral midbrain C4 levels were increased (p < 0.001) versus frontal cortex and striatum in sham-perfused rats, whereas after perfusion there were no differences between brain regions. Removal of cerebrovascular contents decreased C4 by 43% in striatum, 52% in frontal cortex, and 69% in ventral midbrain (all p < 0.01 versus sham-perfused means). These results indicate that C4 in the rat brain can be measured quantitatively by ELISA provided that cerebrovascular proteins are removed by perfusion.     

The effects of vitamin E on antiacid stress ability in juvenile soft-shelled turtles (Pelodiscus sinensis)

We determined the effect of dietary supplementation with vitamin E (0-, 50-, 250-, 500-, 1000- and 5000-mg/kg diet for 4 weeks) on antistress ability in juvenile soft-shelled turtle (Pelodiscus sinensis). Half of the turtles per dose group were treated by acid stress for 24 h. The results showed that phagocytosis of blood cells in the control group significantly decreased after acid stress while the other five groups had no significant changes compared with those of before stress. Serum bacteriolytic activity in the control group and the group supplemented with 50-mg vitamin E/kg diet significantly decreased after acid stress. The other four groups showed no significant differences compared with those before stress. Serum bactericidal activities in all groups notably decreased after acid stress, but the difference of serum bactericidal activity in before and after stress had a decreased tendency from the control group to the highest dose group. Serum cortisol levels in the control group were significantly increased while the other five groups had no notable increases after acid stress. Liver vitamin E levels in all groups had no notable changes compared with those before stress but there was a tendency to decrease after acid stress. These results suggest that acid stress depress immune function and increase serum cortisol levels in turtles while vitamin E alleviate the adverse effects caused by acid stress.     

右美托咪啶对苏醒期小儿血清肌钙蛋白I、C 反应蛋白及补体的影响

目的:探究不同浓度右美托咪啶对小儿麻醉后肌钙蛋白I、C反应蛋白以及补体变化的影响。方法:收集我院下腹股沟斜疝、急性阑尾炎以及肠套叠手术患儿75例,根据治疗方法不同分为三组,每组25例,其中A组实施0.5μg/kg负荷剂量右美托咪啶治疗,B组实施0.25μg/kg负荷剂量右美托咪啶治疗,C组实施右美托咪啶0.2~0.7μg/kg/h持续泵入。对比三组患者苏醒情况、血清肌钙蛋白I、C反应蛋白、补体水平、镇定及躁动评分。结果:手术后,B组自主呼吸恢复时间、气管导管拔管时间、解除监护时间明显缩短,且与其他两组比较,差异有统计学意义(P0.05);与A、C两组相比,B组手术后C反应蛋白及血清肌钙蛋白I水平较低,而C3及C4水平较高,差异具有统计学意义(P0.05);B组镇静、躁动评分明显降低,差异有统计学意义(P0.05)。结论:0.25μg/kg负荷剂量右美托咪啶能够有效减轻患儿术后炎性反应,缓解麻醉后神经损伤,有助于脏器功能的恢复。     

The pH-regulated antigen 1 of Candida albicans binds the human complement inhibitor C4b-binding protein and mediates fungal complement evasion

Candida albicans binds and utilizes human complement inhibitors, such as C4b-binding protein (C4BP), Factor H, and FHL-1 for immune evasion. Here, we identify Candida pH-regulated antigen 1 (Pra1) as the first fungal C4BP-binding protein. Recombinant Pra1 binds C4BP, as shown by ELISA and isothermal titration calorimetry, and the Pra1-C4BP interaction is ionic in nature. The Pra1 binding domains within C4BP were localized to the complement control protein domain 4 (CCP4), CCP7, and CCP8. C4BP bound to Pra1 maintains complement-inhibitory activity. C4BP and Factor H bind simultaneously to Candida Pra1 and do not compete for binding at physiological levels. A Pra1-overexpressing C. albicans strain, which had about 2-fold Pra1 levels at the surface acquired also about 2-fold C4BP to the surface, compared with the wild type strain CAI4. A Pra1 knock-out strain showed ～22% reduced C4BP binding. C4BP captured by C. albicans from human serum inhibits C4b and C3b surface deposition and also maintains cofactor activity. In summary, Candida Pra1 represents the first fungal C4BP-binding surface protein. Pra1, via binding to C4BP, mediates human complement control, thereby favoring the immune and complement evasion of C. albicans.     

Adiponectin binds C1q and activates the classical pathway of complement

The adipose-specific protein adiponectin binds to a number of target molecules, including damaged endothelium and the surface of apoptotic cells. However, the significance of this binding remains unclear. This study demonstrates the binding of purified C1q to recombinant adiponectin under physiological conditions, and the dependence of this upon Ca⁺⁺ and Mg⁺⁺. Binding was enhanced by metaperiodate-mediated destruction of glucosylgalactosyl sugars on adiponectin. Adiponectin was bound by the globular domain of the A chain of collagenase-digested C1q, and C1q binding induced deposition of C4 and C3 through activation of the classical complement pathway. After Western blotting, affinity-purified adiponectin from human serum bound C1q, whereas adiponectin in whole serum did not, unless pre-treated with metaperiodate. These results suggest adiponectin is member of the pattern-recognition family of defence collagens, able to bind target molecules and activate complement. It may therefore play an important role in innate immunity and autoimmune phenomena.     

黄芪对左肾静脉狭窄大鼠肾组织TGF-β1和PAI-1表达的影响

目的观察黄芪对左肾静脉狭窄大鼠肾组织胶原含量及促纤维化因子表达的影响,为临床药物治疗左肾静脉受压综合征提供实验依据。方法将大鼠分为3组,假手术组、模型组和黄芪治疗组。采用左肾静脉不全结扎的方法建立大鼠左肾静脉狭窄模型,用黄芪进行干预,于治疗6周后处死动物。取肾组织行羟脯氨酸含量测定,检测TGF-β1和PAI-1mRNA及蛋白的表达水平。结果模型组大鼠左肾组织羟脯氨酸含量较假手术组增多,TGF-β1和PAI-1的表达增加。黄芪治疗可较少羟脯氨酸含量,减少TGF-β1和PAI-1表达。结论左肾静脉狭窄大鼠可出现肾组织纤维化,黄芪可通过减少TGF-β1和PAI-1的表达来减轻肾组织纤维化损伤。     

中华眼镜蛇蛇毒因子(CVF)对补体系统的作用及与人补作C_3和其他蛇毒抗原性关系

中华眼镜蛇蛇毒经DEAE-Sepharose CL-6B。HPLC等多次柱层析分离出有抗补体及溶血活性的眼镜蛇蛇毒因子(Cobra venom factor,CVF),纯化后的CVF在聚丙烯酰胺凝胶电泳图谱上呈单一区带,分子量为225000—230000,等电点为6.20。用二硫苏糖醇还原经SDS-聚丙烯酰胺凝胶电泳得三类亚基,其分子量总和为237,000。 体外抗补体及溶血试验表明,CVF的作用是通过补体旁路途经使总补体活力下降。双向免疫电泳鉴定,发现CVF与人血清作用后,其中补体成分C_3分子的抗原性发生改变,则表明CVF的作用是通过激活补体成分C_3而发挥的。给豚鼠腹腔注射CVF(0.15ug/g体重)后,其血清总补体水平下降到正常值的3％以下,7天后回升,13天后恢复到正常水平。 单相免疫电泳表明,CVF与人补体C_3抗血清间无任何交叉免疫反应,但人血清与CVF抗血清间有微弱的免疫沉淀反应。另外,CVF的氨基酸组成与人补体C_3也较为相似。鉴定还表明眼镜蛇科中四种蛇毒与CVF抗血清有强烈的免疫沉淀反应,蝰蛇毒及海蛇毒也有免疫沉淀反应,但只有眼镜蛇毒具有抗补体活性。