Use of laser capture microdissection for the assessment of equine lamellar basal epithelial cell signalling in the early stages of laminitis.
Use of laser capture microdissection for the assessment of equine lamellar basal epithelial cell signalling in the early stages of laminitis.
复制标题
使用激光捕获显微切割评估蹄叶炎早期阶段的马层状基底上皮细胞信号传导。
DOI：
10.1111/evj.12283
复制
发表时间：
2015
影响因子：
2.2
通讯作者：
Belknap,JK
中科院分区：
文献类型：
--
作者：
Leise,BS;Watts,MR;Roy,S;Yilmaz,AS;Alder,H;Belknap,JK
进行研究的原因马椎板基底层上皮细胞(LBECs)的黏附异常是导致马椎板炎结构失效的中心事件。研究设计实验研究方法8匹马通过鼻胃管给予过量的含有玉米淀粉混合物的碳水化合物(CHO)。在注射CHO之前，取左前足的板层活检组织(对照[Con])。在发烧时(发育期[DEV])从左后足取活组织，在Obel分级1跛行(OG1)时从右前足取材。用激光捕获显微解剖(LCM)显微镜从冰冻切片中分离板层基底上皮细胞。结果40个基因(22个增加/18个减少)与≤时间相比有显著差异，10 7个基因(57个增加/50个减少)与OG1时间与CON有显著差异。除与细胞外基质组成、稳定性和转换率相关的基因表达显著改变外，炎症相关基因的表达也显著增加。结论与炎症反应和细胞外基质调节相关的信号转导信号在DEV和OG1时段有较强的表达。这些结果表明，LBEC不仅是一个受害者，而且是导致马椎板结构破坏的板层事件的积极参与者。
Reasons for performing studyDysadhesion of laminar basal epithelial cells (LBECs) from the underlying dermis is the central event leading to structural failure in equine laminitis. Although many studies of sepsis‐related laminitis have reported multiple events occurring throughout the lamellar tissue, there is minimal information regarding signalling events occurring specifically in LBECs.ObjectivesTo determine signalling events in the LBECs during the early stages of carbohydrate‐induced laminitis.Study designExperimental study.MethodsEight horses were given an overload of carbohydrate (CHO) consisting of corn starch mixture via nasogastric tube. Prior to administration of CHO, lamellar biopsies were taken from the left forefoot (control [CON]). Biopsies were taken from the left hind foot at the onset of fever (developmental [DEV]) and from the right forefoot at the onset of Obelgrade 1lameness (OG1). Laminar basal epithelial cells were isolated from cryosections using a laser capture microdissection (LCM) microscope. Next generation sequencing (RNA‐seq) was used to identify transcripts expressed in the LBECs for each time point and bioinformatic analysis was performed with thresholds for between group comparisons set at a greater than 2‐fold change and P value ≤0.05.ResultsForty genes (22 increased/18 decreased) were significantly different from DEV time vs. CON and 107 genes (57 increased/50 decreased) were significantly different from OG1 time vs. CON. Significant increases in inflammatory genes were present in addition to significantly altered expression of genes related to extracellular matrix composition, stability and turnover.ConclusionsSignalling related to inflammatory response and extracellular matrix regulation was strongly represented at the DEV and OG1 times. These results indicate that the LBEC is not only a casualty but also an active participant in lamellar events leading to structural failure of the digital lamellae in equine laminitis.