摘要
【目的】从ERK/MAPK通路的角度探讨加味四君子汤对脾虚犬小肠紧密连接蛋白表达的影响。【方法】将24只1岁龄健康比格犬随机均分为对照组、脾虚组、自然恢复组和加味四君子汤组。采用番泻叶药液复制脾虚模型犬,模型构建成功后,脾虚组犬处死采样,对照组和自然恢复组犬饲喂基础日粮,加味四君子汤组犬在基础日粮中添加2%(以生药计)加味四君子汤。试验期14 d。试验结束时各组试验犬解剖采集十二指肠、空肠和回肠,应用免疫组织化学(IHC)、荧光定量PCR、蛋白质印迹法(Western-blot,WB)检测相关指标。【结果】脾虚组试验犬小肠ZO-1和Occludin蛋白阳性表达水平,ZO-1和Occludin mRNA相对表达量,ZO-1、Occludin及ERK/MAPK相关通路蛋白表达量均显著低于对照组试验犬(P<0.01);经加味四君子汤治疗后试验犬上述指标较治疗前均显著升高(P<0.01),趋近对照组水平。【结论】试验犬脾虚证候与小肠紧密连接蛋白的低表达密切相关,小肠机械屏障功能降低;加味四君子汤可以显著提高脾虚犬小肠紧密连接蛋白的表达水平,其调节机制可能是通过活化ERK/MAPK通路来实现。
[Purpose]To determine the effects of supplementary Sijunzi decoction(SJZD) on the expression of tight junction proteins in small intestine of spleen deficiency canines from the perspective of ERK/MAPK pathway.[Method]Twenty-four one-year-old healthy beagle canines were randomly divided into control group, spleen deficiency group, natural recovery group and supplementary SJZD treated group. The senna solution was used to replicate spleen deficiency model canines. After the model was developed, the canines in spleen deficiency group were killed and collecting samples.The canines in the control group and the natural recovery group were fed a basal diet, the canines in supplementary SJZD group were fed a basal diet with 2%(dry herbs) supplementary SJZD. The trial lasted for 14 days. At the end of the experiment, the duodenum, jejunum and ileum were dissected and collected, and the related indexes were detected by immunohistochemistry(IHC), fluorescence quantitative PCR and Western-blot(WB).[Result]The positive expression levels of ZO-1, Occludin proteins, the relative expression values of ZO-1 and Occludin mRNA, and the expression amount of ZO-1, Occludin and ERK/MAPK-related pathway proteins in small intestine of canines in spleen deficiency group were significantly decreased than those in control group(P<0.01). After the treatment with supplementary SJZD, the above indexes of the canines were significantly higher than those before treatment(P<0.01), which were close to the control group.[Conclusion]The syndrome of spleen deficiency was closely related to the low expression of tight junction proteins in the small intestine of the experiment canines, and the mechanical barrier function of the small intestine was reduced. The regulation mechanism may be achieved by activating the ERK/MAPK pathway.
作者
冯士彬
李成
贺濛初
桂雪儿
何嫣婷
李玉
王希春
吴金节
FENG Shibin;LI Cheng;HE Mengchu;GUI Xue’er;HE Yanting;LI Yu;WANG Xichun;WU Jinjie(College of Animal Science&Technology,Anhui Agricultural University,Hefei 230036,China)
出处
《云南农业大学学报（自然科学版）》
CSCD
北大核心
2019年第6期971-979,共9页
Journal of Yunnan Agricultural University:Natural Science
基金
国家重点研发计划项目（2016YFD0501009）
关键词
脾虚
加味四君子汤
紧密连接蛋白
ERK/MAPK通路
spleen deficiency
supplementary Sijunzi decoction(SJZD)
tight junction protein
ERK/MAPK pathway
