鸡球虫病红细胞免疫功能检测

用柔嫩艾美耳球虫人工感染８日龄雏鸡４０只,感染７日后测定红细胞Ｃ３６花环率和ＩＣ花环率,与健康对照组比,Ｃ３６花环率显著降低（Ｐ〈０．０１）,ＩＣ花环率显著升高（Ｐ〈０．０１）,表明鸡球虫病可引起红细胞免疫功能发生改变。     

大弹涂鱼TLR基因分子进化研究及其在鳗弧菌胁迫后的表达分析

大弹涂鱼(Boleophthalmus pectinirostris)是一种生活在潮间带的淤泥滩和红树林等两栖环境中的鱼类,其免疫系统面临比水生生活更大的选择压力。Toll样受体基因(简称TLR)是重要的先天免疫成员,一直是鱼类分子免疫学的研究热点之一。为了探究大弹涂鱼TLR基因是否因为其独特的生活环境而产生适应性进化以及其TLR基因在受到细菌攻击后的免疫应答模式,本研究从大弹涂鱼皮肤转录组中获得了TLR5, TLR8和TLR9完整序列以及TLR3和TLR7部分序列,采用分子生物信息学对大弹涂鱼TLR5, TLR8和TLR9基因序列以及氨基酸序列进行了分析,并根据所构建的系统发育树对5个TLR基因进行了分子进化分析,采用荧光定量PCR方法对大弹涂鱼5个TLR基因的组织表达分布和鳗弧菌攻击后5个TLR基因的免疫应答模式开展了研究。结果显示, TLR5基因全长3071 bp,包括长度为2646 bp的编码区,共编码882个氨基酸;TLR8基因全长3175 bp,包括长度为3033 bp的编码区,共编码1011个氨基酸;TLR9基因全长3398 bp,编码区长度为3093 bp,共编码1031个氨基酸。大弹涂鱼3个TLR基因与其他物种的TLR基因结构相似,具有高度保守性。位点模型结果表明,鱼类TLR3, TLR5和TLR8是高度保守的,而TLR7和TLR9在长期进化过程中产生了适应性进化;而进化枝-位点模型结果表明,为了适应更加复杂多变的两栖环境,大弹涂鱼TLR9基因可能产生了适应性进化。大弹涂鱼5个TLR基因在8个健康组织(肠,眼,肾,肝,脑,肌肉,脾和皮肤)中均有表达,在肝脏和脾脏中的表达量较高。在受到鳗弧菌(Vibrio anguillarum)攻击后的免疫表达模式表明了大弹涂鱼5个TLR基因在应对细菌入侵时起到了重要作用。     

A native picrotoxin-resistant GABA-gated chloride channel receptor subtype in cockroach neurons

Among insect GABA receptors, the GABA-gated chloride channel subtype is insensitive to bicuculline and has been thought to be composed of two populations because of differences in chloride conductance increase, GABA and picrotoxin (PTX) sensitivity. To characterize this possible diversity in GABA-gated chloride channels, electropharmacological experiments were performed on giant interneuron synaptic GABA receptors and on somatic GABA receptors of dorsal unpaired median (DUM) neuron and fast coxal depressor (D_f) motoneuron of the cockroach Periplaneta americana (L). Electrophysiological assays performed at cercal-afferent giant interneuron synapses demonstrated that a biphasic increase in membrane conductance, in response to long-lasting (30 s) neuropilar microapplication of GABA, could be explained by the existence of two GABA-operated chloride channel receptor subtypes. The low stable membrane conductance increase, representing less than 30% of the maximum reached during the early transient phase, was not desensitized quickly. It was reproduced by neuropilar microapplication of cis-4-aminocrotonic acid (CACA) and, in contrast to the fast phase, was not antagonized by bath application of 10⁻⁵ M PTX. Long-lasting (3 min) pneumatic pressure application of GABA on the cell body of motoneuron D_f evoked a fast transient hyperpolarization followed by a slower phase of further hyperpolarization. PTX (10⁻⁵ M ) blocked the fast transient phase and revealed a slow stable hyperpolarization. PTX (10⁻⁴ M ) blocked the major part of the remaining GABA response. The slow hyperpolarization was reproduced by application of CACA. Similar effects of GABA and CACA were recorded on DUM neuron cell bodies. All of these observations are consistent with the possible existence of two GABA-gated chloride channel subtypes in the insect CNS. © 1999 Society of Chemical Industry     

Bicyclophosphorothionate antagonists exhibiting selectivity for housefly GABA receptors

2,6,7-Trioxa-1-phosphabicyclo[2.2.2]octane 1-sulfides (bicyclophosphorothionates) with various C_1–4 alkyl groups at the 3- and 4-positions were synthesized and tested for their ability to compete with [³H]4′-ethynyl-4-n-propylbicycloorthobenzoate (EBOB), a non-competitive antagonist of γ-aminobutyric acid (GABA) receptors, for specific binding to rat-brain and housefly-head membranes, and for their insecticidal activity against houseflies. Among the 3,4-substituted analogues, 20 compounds were selectively active for housefly GABA receptors versus rat GABA receptors. The 3-alkyl groups of C₃ length and the 4-alkyl groups of C₄ length were tolerated in housefly receptors, whereas such bulky substituents were deleterious in rat receptors. The 4-isobutyl-3-isopropyl analogue was the most potent in housefly receptors (IC₅₀ = 45.2 nM ), and tert-butylbicyclophosphorothionate (TBPS), with the 4-tert-butyl group and no 3-substituent, was the most potent in rat receptors (IC₅₀ = 62.2 nM ). Their receptor selectivities (rat IC₅₀/housefly IC₅₀) were 52 and 0.038, respectively. The insecticidal activity (LD₅₀) of 20 active analogues was well correlated with their potency (IC₅₀) in inhibiting [³H]EBOB binding to housefly-head membranes (r = 0.93). The results obtained in the present study indicate that the introduction of appropriate alkyl groups into the 3- and 4-positions of bicyclophosphorothionate leads to non-competitive antagonists with increased affinity and selectivity for housefly ionotropic GABA receptors versus rat GABA_A receptors. © 1999 Society of Chemical Industry     

Activation of TGR5 reduces high glucose-induced cardiomyocyte hypertrophy by inhibiting CaN/NFAT3 signaling

AIM: To investigate the role of G-protein-coupled bile acid receptor 1(GPBR1; also known as TGR5) activation in high glucose-induced cardiomyocyte hypertrophy and calcineurin (CaN)/nuclear factor of activated T-cells 3 (NFAT3) signaling.METHODS: Primarily cultured mouse cardiomyocytes were used in the study. The cell surface areas of the cardiomyocytes were measured by an image analysis system. The cell protein content was detected by BCA method. The expression of TGR5, CaN and NFAT3 at mRNA and protein levels was determined by RT-PCR and Western blot.RESULTS: The mouse cardiomyocytes were successfully cultured. High glucose significantly induced the increases in the cell surface area, the cell protein content and the expression of CaN and NFAT3 (P<0.05) in the cardiomyocytes. TGR5 activation or a CaN antagonist cyclosporin A inhibited high glucose-induced cardiomyocyte hypertrophy and the expression of CaN and NFAT3 (P<0.05). These effects of TGR5 activation were abolished by TGR5 gene interference (P<0.05).CONCLUSION: TGR5 activation reduces high glucose-induced cardiomyocyte hypertrophy by inhibiting CaN/NFAT3 signaling.     

Expression profiles and functional prediction of ionotropic receptors in Asian corn borer,Ostrinia furnacalis (Lepidoptera: Crambidae)

Genes involved in chemosensation are essential for odorant-mediated insect behaviors. Odorant receptors (ORs) bind and respond to pheromones and plant volatiles, regulating insect behaviors such as mating and host-plant selection, while ionotropic receptors (IRs), which are present at lower levels in insects than ORs, influence ion channels, especially in agricultural pests. Asian corn borer, Ostrinia furnacalis, is the main pest of maize that causes huge economic losses in Asia. Twenty-one OfurIRs have been identified, but none has been characterized. In this study, tissue-specific expression profiling, phylogenetic analysis, and electroantennography (EAG) analysis were applied to characterize the evolution, expression, and the potential function of OfurIRs. It was found that 20 OfurIRs were highly expressed in the antennae, except for OfurIR75p3, whereas 10 and nine OfurIRs were highly expressed in the proboscis and genitalia, respectively, indicating that these OfurIRs were functionally associated with feeding and oviposition. EAG results showed that seven acids elicited responses in the antennae of O. furnacalis and that 2-oxopentanoic acid displayed a significant female-biased response. Combined with the phylogenetic analysis, 10 OfurIRs in clade 4 were roughly predicted to be candidate receptors for 2-oxopentanoic acid and other tested acids. These results provide basic information about OfurIRs and may help advance the knolwedge on the olfactory system of O. furnacalis.     

外源性γ-氨基丁酸抵抗仔猪氧化应激的效果及机制

【目的】 研究外源性γ-氨基丁酸（GABA）抵抗仔猪氧化应激的作用效果,及海马神经元GABA受体调节凋亡信号通路在其中可能的介导作用,为GABA作为动物应激调节剂的应用提供科学依据。【方法】 基于成功构建的仔猪活体和大鼠海马神经元氧化应激模型,考察外源性GABA对仔猪血清及海马组织中氧化/抗氧化相关指标、仔猪日增重、脑海马GABA受体以及大鼠海马神经元中GABA受体及凋亡信号通路相关基因表达水平的影响。【结果】 低、中、高浓度的GABA灌喂组（LD+OS; MD+OS;HD+OS）仔猪血清的MDA含量均极显著低于氧化应激（OS）组(P<0.01),而GSH水平均极显著高于OS组(P<0.01),同时HD+OS 组仔猪血清T-AOC水平极显著高于OS组和对照组(P<0.01);且高浓度(100 mg·kg^-1 BW)GABA降低仔猪血清MDA含量和增加GSH水平的幅度均高于低浓度(20 mg·kg^-1 BW)和中浓度(60 mg·kg^-1 BW)GABA。因此,后续研究仅考察100 mg·kg^-1 BW GABA的作用效果及其抗氧化应激的机制。OS组仔猪0-7、8-14和0-28日龄的日增重极显著低于对照组(P<0.01),而HD+OS组0-7、8-14和0-28日龄仔猪的日增重极显著高于OS组(P<0.01);OS组仔猪15-28日龄的日增重显著低于对照组(P<0.05),HD+OS组15-28日龄的日增重极显著高于OS组(P<0.01)。以上结果表明,100 mg·kg^-1 BW GABA的灌喂极显著地增加了仔猪的日增重。对照组、OS组和HD+OS组在前、中、后期的腹泻率均无显著差异(P>0.05)。氧化应激组海马组织的MDA含量极显著高于对照组和HD+OS组 (P<0.01),而T-AOC和GSH的水平均极显著低于另外两组(P<0.01),表明GABA能提高仔猪海马组织抗氧化能力。HD+OS组的海马组织GABA_A和GABA_B受体水平均极显著高于对照组和氧化应激组(P<0.01),表明GABA提高了海马组织GABA_A和GABA_B的水平。氧化应激组脑海马Bcl-2蛋白水平显著低于对照组(P<0.05),Bax和Caspase-3蛋白水平极显著高于对照组(P<0.01)。HD+OS组的Bcl-2蛋白水平极显著高于氧化应激组（P<0.01）,Bax蛋白水平极显著低于氧化应激组（P<0.01）,Caspase-3蛋白水平显著低于氧化应激组（P<0.05）。与此一致的是,氧化应激组、GABA+OS+Picrotoxin组和GABA+OS+CGP54626组大鼠海马神经元的Bax和Caspase-3蛋白水平均显著高于对照组和GABA+OS组（P<0.05）,表明GABA缓解了氧化应激状态下海马神经元的损伤,而GABA受体抑制剂的添加阻挡了GABA的抗应激损伤作用。【结论】 GABA降低了仔猪海马的氧化应激水平,而GABA抗应激的作用机制可能与其降低凋亡蛋白基因的表达相关,而GABA_A和GABA_B受体介导了该过程。