Staphylococcus aureus (S. aureus) is one of the most infamous and widespread bacterial pathogens, causing a hard-to-estimate number of uncomplicated skin infections and probably hundreds of thousands to millions of more severe, invasive infections globally per year. S. aureus may also be acquired from animals, especially in the livestock industry. The interaction mechanism of host and S. aureus has significance for finding ways to against S. aureus infection and control inflammatory response of host, while the molecular biological activities after S. aureus infection, particular in inflammatory and immune cells are not fully clear. The present study aimed to explore whether pattern recognition receptors (PRRs) mediate prostaglandin D2 (PGD2) synthesis and PGD2 participates in the regulation of inflammatory response in macrophages during S. aureus infection or synthetic bacterial lipopeptide (Pam2CSK4) stimulation. PGD2 secretion level was enhanced by mice peritoneal macrophages infected with the S. aureus. The results indicated that PGD2 secretion was impaired in S. aureus infected-macrophages from toll-like receptors 2 (TLR2)-deficient and NLR pyrin domain-containing 3 (NLRP3)-deficient mice. PGD2 synthetase (hematopoietic PGD synthase, HPGDS) inhibitors could reduce the activation of macrophage mitogen-activated protein kinase (MAPK)/nuclear factor-κ-gene binding (NF-κB) signaling pathways. HPGDS inhibition impaired cytokines (TNF-α, IL-1β, IL-10 and RANTES) secretion and macrophage phagocytosis during S. aureus infection. In addition, inhibition of endogenous PGD2 synthesis was unable to affect the TLR2 and NLRP3 expression in S. aureus-infected macrophages. Taken together, macrophage PGD2 secretion after S. aureus infection depended on receptors TLR2 and NLRP3, and the induced PGD2 participated in the regulation of inflammatory response in S. aureus-infected macrophages. Interestingly, it was found that exogenous PGD2 down-regulated the cytokines secretion and had no effect on phagocytosis in the S. aureus-infected macrophages.

布鲁菌病(简称布病)是最广泛的细菌性人畜共患病之一,会导致家畜大规模感染从而造成重大经济损失,其在人群中的传播也给人类社会造成危害.布鲁菌可以通过吸入、皮肤擦伤、摄入或黏膜直接接触等多种方式进入宿主细胞,是一种细胞内病原体,可以逃避宿主免疫反应从而在细胞内增殖,引起持续感染.本文主要从布鲁菌病的病原体特征、临床症状、检测技术、疫苗研发及防治净化等方面进行阐述,以期为布病的预防控制提供理论基础.

为探究枯草芽孢杆菌细胞壁能否对绵羊瘤胃上皮细胞(ovine ruminal epithelial cells,ORECs)中绵羊β-防御素-1(sheep β-defesin-1,SBD-1)的表达产生影响,先利用反复冻融和超声波破碎相结合的方法破碎枯草芽孢杆菌并成功收集其细胞壁,将不同浓度(0、25、50、100、200、400 μg·mL-1)的枯草芽孢杆菌细胞壁与ORECs共培养8h后,采用荧光定量PCR和酶联免疫吸附测定(ELISA)方法筛选出枯草芽孢杆菌细胞壁诱导SBD-1表达的最佳刺激浓度,最后用该浓度条件下的枯草芽孢杆菌细胞壁对瘤胃上皮细胞进行不同时间(2、4、8、12、24 h)的刺激,采用荧光定量PCR和ELISA方法筛选出枯草芽孢杆菌细胞壁诱导SBD-1表达的最佳刺激时间.结果 显示:枯草芽孢杆菌细胞壁成功制备,不同浓度(0、25、50、100、200、400 μg· mL-1)的枯草芽孢杆菌细胞壁对ORECs均无毒性作用,并能极显著促进ORECs中SBD-1 mRNA和蛋白的表达(P＜0.01),以浓度为50 μg·mL-1的枯草芽孢杆菌细胞壁刺激ORECs 12 h时SBD-1 mRNA和蛋白的表达量达到最大且与对照组呈极显著差异(P＜0.001).研究表明,枯草芽孢杆菌细胞壁刺激ORECs诱导SBD-1表达的最佳刺激浓度为50 μg·mL-1,最佳刺激时间为12 h.本研究为枯草芽孢杆菌细胞壁诱导调节SBD-1机制的研究提供理论依据.

[目的]建立永生化绵羊瘤胃成纤维细胞系,为基础研究和动物病毒的研究提供稳定的体外细胞模型.[方法]采用组织块贴壁法培养绵羊瘤胃成纤维细胞(ovine ruminal fibroblasts cells,ORFCs),利用PEI试剂将带有hTERT基因的pCI-neo-hTERT质粒转染ORFCs,经G418筛选得到阳性克隆细胞.采用RT-PCR、Western Blot检测F15和F35代阳性克隆细胞的hTERT基因的转录和表达情况;利用血球计数法绘制原代ORFCs和F35代阳性克隆细胞的生长曲线比较其增殖能力.[结果]将hTERT基因成功转入ORFCs并稳定表达;阳性克隆细胞的增殖能力显著高于原代ORFCs.[结论]该试验成功建立了永生化绵羊瘤胃成纤维细胞系.

以体外培养的绵羊瘤胃上皮细胞(ovine rumen epithelial cells,ORECs)为试验模型,旨在探究灭活枯草芽孢杆菌对ORECs绵羊β-防御素-1(sheepβ-defesin-1,SBD-1)表达的影响.首先用不同浓度的灭活枯草芽孢杆菌(107、108、109、1010、1011 CFU/mL)诱导ORECs 8 h,通过荧光定量PCR(qPCR)和ELISA的方法检测ORECs SBD-1表达量在mRNA水平和蛋白水平的变化,确定灭活枯草芽孢杆菌诱导ORECs SBD-1表达最高的菌液浓度为最佳刺激浓度.以该浓度的灭活枯草芽孢杆菌诱导ORECS不同时间(2 h、4 h、8 h、12 h),通过qPCR和ELISA的方法检测ORECs SBD-1表达量在mRNA水平和蛋白水平的变化,从而筛选出诱导SBD-1表达的最佳时间.结果表明,108～1011 CFU/mL的灭活枯草芽孢杆菌可以显著的促进ORECs SBD-1的表达,其中灭活枯草芽孢杆菌的浓度为1010 CFU/mL时诱导ORECs 8h时SBD-1 mRNA表达量最高,与对照组相比差异极显著(P<0.01);SBD-1蛋白表达量以1010 CFU/mL灭活枯草芽孢杆菌诱导ORECs 12 h时达到最大,与对照组相比差异极显著(P<0.01).结果证明了热灭活后的枯草芽孢杆菌仍可以诱导SBD-1表达,其诱导SBD-1表达的有效成分并未被热灭活,为后续确定枯草芽孢杆菌诱导SBD-1表达的有效成分的研究提供了理论基础.

为了探索膜受体Dectin-2和TLR-2在酿酒酵母甘露聚糖诱导绵羊瘤胃上皮细胞(ovine ruminal epithelial cells,ORECs)β-防御素-1(sheep β-defensin-1,SBD-1)表达过程中的作用.本研究首先利用免疫组化、RT-PCR和Western blot方法确定Dectin-2是否在ORECs内表达;然后采用qPCR和Western blot方法检测甘露聚糖刺激ORECs后Dectin-2和TLR-2的表达变化;最后用qPCR和ELISA方法检测不同质量浓度(0.1,1.0,10.0 mg/L)的Dectin-2和TLR-2特异性封闭抗体对甘露聚糖诱导SBD-1表达的影响.结果 显示:ORECs内表达Dectin-2,且甘露聚糖刺激ORECs后Dectin-2和TLR-2的表达量均显著增加(P＜0.01);同时甘露聚糖可以诱导SBD-1的表达(P＜0.01),但是该表达过程可以被不同质量浓度的Dectin-2封闭抗体和TLR-2封闭抗体所抑制(P＜0.05),且随着抗体浓度的增加,这种抑制作用越明显.此外,在封闭抗体质量浓度相同时,Dectin-2封闭抗体比TLR-2封闭抗体抑制SBD-1表达的作用更明显(P＜0.05).结果 表明,酿酒酵母甘露聚糖诱导SBD-1的表达与Dectin-2和TLR-2受体有关,但是Dectin-2在此过程中发挥更主要的作用.

In this study, we investigated whether β-glucan from Saccharomyces cerevisiae exerts beneficial effects on mucosal immunity in an ovine ruminal explant (ORE) model. Once the ORE model was established, viability was assessed through histological change, E-cadherin expression, CK-18 and Ki-67 distribution. Then, the OREs were co-cultured with β-glucan, following which, gene and protein expression levels of sheep β-defensin-1 (SBD-1), pro-inflammatory interleukin (IL)-6, and anti-inflammatory IL-10 were detected using quantitative real-time polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA). Hematoxylin & eosin staining, qPCR, and immunohistochemistry showed that the overall ORE structure was intact after 96 hours in culture, but explants cultured for more than 24 hours showed epithelial degradation. Therefore, we performed the follow-up test within 24 hours. qPCR and ELISA revealed that the gene and protein expression levels of SBD-1, IL-6, and IL-10 in the OREs significantly increased (P < 0.05) after treatment with β-glucan compared with controls. This study identified the feasibility and optimal conditions of ORE culture and demonstrated that β-glucan activates SBD-1, IL-6, and IL-10 secretion in OREs to promote mucosal immunity.

建立绵羊瘤胃上皮细胞(ovine ruminal epithelial cells,ORECs)培养体系为体外试验模型,旨在探索酿酒酵母培养物和提取物对ORECs中绵羊β-防御素-1(sheepp-defesin-1,SBD-1)表达效果的影响.首先,分别用200 mg/L酿酒酵母培养物和提取物诱导ORECs不同时间段(0,6,12,18,24,30 h)后,经qPCR扩增反应检测SBD-1 mRNA表达量,以分别确定酿酒酵母培养物和提取物诱导SBD-1 mRNA表达最高的刺激时间;然后用不同质量浓度梯度(0,100,200,300,400 mg/L)酿酒酵母培养物和提取物分别诱导ORECs最佳时间段后,同样地采用qPCR扩增技术检测SBD-1 mRNA表达量,从而筛选出诱导SBD-1mRNA表达最高的酿酒酵母培养物浓度和酿酒酵母提取物浓度.结果 显示,酿酒酵母培养物和提取物均可诱导SBD-1 mRNA表达.当酿酒酵母培养物质量浓度为400 mg/L、酿酒酵母提取物质量浓度为200 mg/L分别刺激ORECs 6 h时,ORECs SBD-1 mRNA表达量分别达到最高,且均极显著高于对照组(P＜0.01).最后在相同试验条件下,分别使用最佳诱导质量浓度的酿酒酵母培养物(即400 mg/L)和酿酒酵母提取物(即200 mg/L)诱导ORECs最佳时间(即6 h),比较两者诱导ORECs SBD-1 mRNA表达效果,qPCR结果显示酿酒酵母提取物诱导SBD-1 mRNA表达显著高于酿酒酵母培养物(P＜0.05).结果 表明,酿酒酵母培养物和提取物均能够诱导ORECs SBD-1表达,400 mg/L酿酒酵母培养物和200 mg/L酿酒酵母提取物分别刺激ORECs 6 h后SBD-1表达量最高,且酿酒酵母提取物诱导效果优于酿酒酵母培养物.

Ovine ruminal epithelial cells (ORECs) not only have a physical barrier function but also can secrete host defence peptides (HDPs), such as sheep β-defensin-1 (SBD-1). As a feed additive, Saccharomyces cerevisiae can enhance the host’s innate immunity. β-glucan, a cell wall component of Saccharomyces cerevisiae , can stimulate innate immune responses and trigger the up-regulation of SBD-1 in ORECs. The signaling mechanisms involved in β-glucan-induced SBD-1 expression are not completely understood. The aim of this study was to identify the receptors and intracellular pathways involved in the up-regulation of SBD-1 induced by β-glucan. ORECs were cultured, and the regulatory mechanisms of β-glucan-induced up-regulation of SBD-1 were detected using quantitative real-time PCR (qPCR), enzyme-linked immunosorbent assay (ELISA), and western blotting. TLR-2 and MyD88 knockdown or inhibition attenuated β-glucan-induced SBD-1 expression. We also showed that inhibition of MAPK and NF-κB pathways significantly reduced β-glucan-induced SBD-1 expression. These results demonstrate that β-glucan-induced SBD-1 expression is TLR-2-MyD88-dependent and may be regulated by both MAPK and NF-κB pathways. Since NF-κB inhibition had a greater effect on the down-regulation of β-glucan-induced SBD-1 expression, the NF-κB pathway may be the dominant signaling pathway involved in the regulation of defensin expression. Our studies demonstrate that β-glucan-induced SBD-1 expression is mediated through the TLR-2-MyD88-NF-κB/MAPK pathway. Our results would contribute to the understanding of immunological modulations in the gastrointestinal tract triggered by probiotic yeast cell wall components.

The rumen epithelium of sheep serves as an immune interface with the environment and secretes antimicrobial peptides with bactericidal function against various pathogens. Sheep beta-defensin-1 (SBD-1), an antimicrobial peptide, is secreted from ovine ruminal epithelial cells (OREC) in response to microbial infections. Mannan, the main component of the Saccharomyces cerevisiae cell wall can stimulate innate and regulatory immune responses that could improve the gastrointestinal environment. We aimed at investigating the effects of mannan on SBD-1 expression and the downstream signaling pathways stimulated in OREC. We cultured OREC; assessed the effects of mannan on SBD-1 expression by qPCR and ELISA; and then investigated the underlying signaling pathways using qPCR, ELISA, Western blotting, immunohistochemistry, and immunohistofluorescence. Interestingly, mannan markedly upregulated SBD-1 expression in a concentration- and time-dependent manner. Dectin-2 Mouse mAb, Syk specific inhibitor R406, and specific inhibitors of the p38, ERK1/2, JNK, and NF-κB pathways attenuated mannan-induced SBD-1 expression to varying degrees. These results demonstrate that SBD-1 is upregulated by mannan via the Dectin-2-Syk axis, and this is regulated to a large extent through the mitogen-activated protein kinase (MAPK) p38 and less so through the ERK1/2 and JNK or the NF-κB pathway. Our findings highlight the immunomodulatory effects of mannan on OREC in terms of mannan-induced SBD-1 expression.

旨在研究酿酒酵母细胞壁对体外培养绵羊瘤胃上皮细胞β-防御素-1 (sheep beta-defensin-1,SBD-1)表达的影响及其调控途径,为揭示酿酒酵母细胞壁对机体免疫的调控机制提供理论依据.本研究将不同质量浓度的酿酒酵母细胞壁提取物(0,25,50,100,200,400 mg/L)作用于绵羊瘤胃上皮细胞不同时间(0,2,4,8,12,24 h)后,检测上皮细胞SBD-1和Toll样受体-2(TLR2)mRNA表达水平,并进一步通过TLR2阻断试验来证实TLR2是否介导酵母细胞壁促进SBD-1表达.结果 表明,不同浓度的酵母细胞壁刺激瘤胃上皮细胞时,随着细胞壁浓度的增加SBD-1mRNA表达量呈先升高后降低的趋势,且酿酒酵母细胞壁质量浓度为200 mg/L刺激12h时,SBD-1 mRNA表达量达到峰值,与对照组和其他时间组相比差异显著(P＜0.05);用质量浓度为200 mg/L的酿酒酵母细胞壁刺激绵羊瘤胃上皮细胞不同时间,同样在12 h时,TLR2的mRNA表达量达到最大;阻断试验表明TLR2可介导SBD-1的表达.本研究结果表明酿酒酵母细胞壁可促进绵羊瘤胃上皮细胞SBD-1的表达,且质量浓度200 mg/L时,刺激瘤胃上皮细胞12 h时SBD-1的表达量达到最高,而TLR2参与该调控过程.

<span id="ChDivSummary" name="ChDivSummary" class="abstract-text">本研究旨在建立绵羊瘤胃上皮细胞（Ovine rumen epithelial cells,ORECs）的分离培养、冻存和复苏方法,为反刍动物瘤胃功能的研究和相关瘤胃疾病的发病机制研究提供功能性的细胞模型。以绵羊瘤胃为研究对象,采用胰蛋白酶连续消化法对瘤胃组织进行不同时间（40、30、20、10、10、8和5 min）的消化,并对每次消化后的瘤胃组织和消化液分别进行H.E染色分析和显微镜观察,以确定细胞开始收集及终止消化的时间,最后将收集的瘤胃上皮细胞进行原代培养。在传代培养时,运用差时消化法和差速贴壁法对ORECs进行纯化,并从细胞形态学、细胞生长曲线、角蛋白18（CK-18）免疫荧光染色和上皮细胞标志物E-钙黏蛋白和碱性磷酸酶基因表达等方面对ORECs进行鉴定。然后设置不同体积配方的冻存液,将传代细胞进行冻存,通过检测复苏后的细胞活力和24 h贴壁率确定最佳冻存液配方。结果表明:第4次消化后就基本消化到瘤胃组织的棘层,此时即可开始收集细胞;第7次消化完后瘤胃上皮的基底层细胞基本被消化掉,此时即可终止消化细胞。经纯化后传代的细胞呈现均一的"铺路石"形态,生长曲线明显呈"S"形,且经CK-18免疫荧光染色后鉴定为阳性,同时PCR检测到上皮细胞特异性标志蛋白E-钙黏蛋白和碱性磷酸酶均有表达。此外,ORECs经不同体积配方冻存液冻存复苏后发现,冻存液配方为V_FBS∶V_DMEM∶V_DMSO=9∶0∶1时ORECs的细胞活力及贴壁率最高,且传代后的细胞生长状况良好。因此本试验成功建立了ORECs的体外分离培养、冻存和复苏方法。</span>

据报道,全球每年仅因奶牛乳腺炎造成的经济损失就高达350亿美元,我国每年因奶牛乳腺炎造成的经济损失也达15亿人民币[1].奶牛乳腺炎是由革兰阴性菌和阳性菌以及真菌引起的在奶牛中最常见也是最有经济破坏力的疾病,对产奶质量和数量都有较大影响[2].而在兽医治疗过程中使用抗生素和牲畜淘汰仍为治疗奶牛乳腺炎的主要手段.但也由于乳腺炎治疗过程中抗生素的大量使用对牛奶质量造成了显著影响,如乳制品中抗生素残留,会对饮用这类问题牛奶的人造成生理伤害,从而造成更大范围内的经济损失.到目前为止,尽管奶牛乳腺炎已经得到了较为有效的控制,但它仍是现阶段全球乳业所面临的主要挑战之一.

β-防御素是机体的内源性抗菌肽.酿酒酵母(Saccharomyces cerevisiae)甘露聚糖能够诱导绵羊(Ovis aries)瘤胃上皮细胞(ruminal epithelial cells,RECs)β-防御素-1 (sheepβ-defensin-1,SBD-1)的表达,但是诱导机制尚不明确,限制了甘露聚糖制剂的开发利用.为了探讨丝裂原活化蛋白激酶(mitogenactivated protein kinase,MAPK)信号传导通路(p38,ERK1/2,JNK)和核因子κB (nuclear factor κB,NF-κB)通路是否参与甘露聚糖诱导绵羊RECs SBD-1的表达,首先利用qPCR和Western blot方法检测甘露聚糖对p38、ERK1/2、JNK和NF-κB表达的影响;然后用甘露聚糖分别刺激RECs 5、15、30、45和60 min后,通过Western blot检测p38、ERK 1/2、JNK、IκB和p65的磷酸化水平;最后采用qPCR和ELISA方法检测p38、ERK1/2、JNK和NF-κB的特异性抑制剂对甘露聚糖诱导SBD-1表达的影响.结果 显示,甘露聚糖刺激使RECs中p38、ERK1/2、JNK和NF-KB的mRNA水平以及p38、JNK、ERK1/2、IκB和p65的磷酸化水平均显著提高(P＜0.01);同时,甘露聚糖刺激RECs不同时间可以使p38、ERK1/2、JNK、IκB和p65发生磷酸化;此外,p38、ERK1/2、JNK和NF-κβ的抑制剂均能极显著降低甘露聚糖诱导SBD-1的表达(P＜0.01),且p38抑制剂的抑制效果最明显.上述结果提示,酿酒酵母甘露聚糖诱导绵羊RECs SBD-1表达的过程可能由p38、ERK1/2、JNK和NF-κB信号通路共同调节,并且p38信号通路可能发挥更主要的作用.该研究探讨了甘露聚糖诱导防御素表达的机理,为解释甘露聚糖促免疫功能提供了新线索,也为更好地开发和利用甘露聚糖制剂提供了参考依据.

为了探索酿酒酵母β-葡聚糖诱导绵羊瘤胃上皮细胞(ovine ruminal epithelial ceils,ORECs)β-防御素-1(sheep β-defensin-1,SBD-1)表达过程中膜受体Dectin-1和TLR-2可能存在的作用.本研究首先利用免疫组化、RT-PCR、免疫荧光和Western blot等方法检测Dectin-1是否在ORECs内表达,并且采用qPCR和Western blot方法对β-葡聚糖刺激ORECs后细胞膜受体Dectin-1和TLR-2的表达变化进行检测.而后用不同浓度的Dectin-1阻断剂昆布多糖或TLR-2特异性封闭抗体分别预处理ORECs后,采用qPCR和ELISA方法检测SBD-1的表达变化,以确定β-葡聚糖诱导SBD-1表达过程中Dectin-1和TLR-2的参与情况.结果 显示:1)绵羊瘤胃组织及ORECs内存在Dectin-1表达,且β-葡聚糖刺激ORECs后Dectin-1和TLR-2的表达水平显著增加(P＜0.05);2)不同浓度的昆布多糖和TLR-2封闭抗体均可以极显著降低β-葡聚糖诱导SBD-1的表达(P＜0.01),且随着阻断剂和封闭抗体浓度的增加,其抑制SBD-1表达的作用越明显.结果 表明,Dectin-1在绵羊瘤胃组织及ORECs内均表达,并且酿酒酵母β-葡聚糖在ORECs中诱导SBD-1的表达是由Dectin-1和TLR 2介导产生的.

近年来,随着国际市场自由贸易的冲击,国内更多的养殖企业大力引进或培育产肉性能突出的肉用绵羊,如杜柏、萨福克等,以获取更可观的经济利润.据调查,很多肉羊养殖基地陆续出现直肠脱垂或直肠黏膜水肿外脱的病例,由于未达到有效控制,导致病羊死亡或提前淘汰,给养殖企业带来严重的经济损失,给养殖工作者带来一定困扰.

The ruminal mucosal epithelium can secrete defensins, which play a key role in innate and adaptive immunity and are considered potential replacements for antibiotics. Of these, sheep β-defensin-1 (SBD-1) is one of the most potent molecules produced by ovine ruminal epithelial cells (ORECs). β-glucan, safe and effective immune activators, can stimulate innate and adaptive immune responses. Here we examined whether β-glucan from Saccharomyces cerevisiae can induce SBD-1 expression in ORECs, as well as the underlying mechanism. First, ORECs were cultured, and quantitative real-time PCR (qPCR) and enzyme-linked immunosorbent assay (ELISA) were used to study the effects of different β-glucan concentrations. Then western blotting, immunohistochemistry, and immunocytofluorescence were performed to investigate the regulatory mechanism of β-glucan-induced SBD-1 upregulation. We show that β-glucan can induce the release of SBD-1 from ORECs; the highest SBD-1 mRNA and protein expression was achieved after treatment with 10 μg/mL at 2 and 4 h. Moreover, β-glucan-induced SBD-1 production was mediated by the activation of dendritic-cell-associated C-type lectin 1 (Dectin-1) receptors, Syk, and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB). These findings highlight the immunomodulatory effects of β-glucan on ORECs.

建立绵羊瘤胃外植体(ovine ruminal explants,OREs)模型,并根据组织学变化和E-钙黏蛋白(E-cadherin)的表达及CK-18和Ki-67的分布评估其活力.培养方案建立后,采用qPCR和ELISA方法检测β-葡聚糖刺激OREs对绵羊β-防御素-1(β-defensin-1,SBD-1)mRNA和蛋白表达水平的影响.HE染色,qPCR和免疫组织化学结果显示培养基内不添加血清培养24 h内的OREs的总体结构较完整,且具有活性.qPCR和ELISA显示,与对照组相比用β-葡聚糖刺激OREs后显著增加SBD-1的mRNA和蛋白表达(P＜0.05).本试验确定了OREs体外培养的可行性和最佳条件,并证明β-葡聚糖可以激活瘤胃中抗菌肽SBD-1的分泌.

为研究Ghrelin在马鹿体内可能表达的器官,通过反转录-聚合酶链式反应(RT-PCR)和实时荧光定量PCR(Real-time quantitative PCR,qPCR)方法检测马鹿的舌、食管、瘤胃、网胃、瓣胃、皱胃、十二指肠、空肠、回肠、盲肠、结肠、直肠、肝、胰、颌下腺、腮腺、气管、肺、肾、输尿管、膀胱、卵巢、输卵管、子宫、阴道、大脑、下丘脑、垂体、松果体、甲状腺、肾上腺、心肌、心内膜、脾、淋巴结和骨骼肌中Ghrelin表达情况.结果显示,Ghrelin在皱胃内的表达量明显高于其他器官(P＜0.05);胰、子宫、卵巢、十二指肠和食管次之,与其余30种器官的表达量相比差异也显著(P＜0.05);其余器官内的表达量相对较少.结论:Ghrelin在所检测的36种器官内均有表达.