槲皮素干预猪流行性腹泻病毒感染幼龄仔猪回肠蛋白质组的研究

Effect of quercetin on ileal proteome of young piglets infected with porcine epidemic diarrhea virus

试验基于蛋白质组分析探究槲皮素抑制幼龄仔猪肠道内猪流行性腹泻病毒(PEDV)增殖的机制。选取18头体重相近的7日龄健康断奶仔猪,随机分为3组,每组6个重复,每个重复1头猪。试验期11 d,第0~3 d为预试期;第4~10 d,槲皮素+PEDV组仔猪口腔灌服10 mg/kg BW溶解在人工乳中的槲皮素,对照组和PEDV组仔猪灌服同体积的人工乳;第8 d,PEDV组和槲皮素+PEDV组的仔猪灌服10^(4.5)TCID_(50)的PEDV,对照组仔猪灌服等体积的PBS。结果显示:与对照组相比,PEDV上调了感染仔猪十二指肠、空肠、回肠、结肠的PEDV-N基因的相对表达量(P<0.05)。与PEDV组相比,槲皮素抑制了仔猪空肠、回肠PEDV-N基因的相对表达量(P<0.05)。与对照组相比,PEDV组仔猪肠道(S)-3-氨基-2-甲基丙酸转氨酶(ABAT)表达显著上调(P<0.05),精氨琥珀酸合酶(ASS1)、鸟氨酸氨基转移酶(OAT)、Y+L氨基酸转运蛋白(y^(+)LAT1)、兴奋性氨基酸转运载体1(EAAC1)、b(0,+)型氨基酸转运蛋白1(b^((0,+))AT)、高亲和力阳离子氨基酸转运蛋白1(CAT-1)表达显著下调(P<0.05)。与PEDV组相比,槲皮素+PEDV组仔猪ABAT、ASS1、OAT、y^(+)LAT1、EAAC1、b^((0,+))AT、CAT-1的表达显著上调(P<0.05)。研究表明,槲皮素可能通过调控肠道氨基酸的转运和代谢抑制PEDV在仔猪体内的增殖。

The experiment was based on proteomic analysis to explore the mechanism of quercetin inhibiting the proliferation of porcine epidemic diarrhea virus(PEDV) in intestinal tract of young piglets. A total of 18 seven-day-old healthy weaned piglets with similar body weight were selected and randomly divided three groups, with six replicates in each group and one pig in each replicate. The experimental period was 11 d, and 0~3 d was pre-test period. On 4~10 d, the piglets in quercetin+PEDV group were orally administered 10 mg/kg BW quercetin dissolved in artificial milk, the piglets in control group and PEDV group were fed with the same volume of artificial milk. On 8 d, the piglets in PEDV group and quercetin+PEDV group were fed with 10^(4.5)TCID_(50) of PEDV, and piglets in control group were fed with the same volume of PBS. The results showed that compared with control group, PEDV increased the relative expression of PEDV-N gene in duodenum, jejunum, ileum and colon of infected piglets(P<0.05). Compared with PEDV group, quercetin inhibited the relative expression of PEDV-N gene in jejunum and ileum of piglets in quercetin+PEDV group(P<0.05). Compared with control group, the intestinal ABAT expression of piglets in PEDV group was significantly increased(P<0.05), and the expressions of ASS1, OAT, y^(+)LAT1, EAAC1, b^(0,+)AT, CAT-1 were significantly reduced(P<0.05). Compared with PEDV group, the expressions of ABAT, ASS1, OAT, y^(+)LAT1, EAAC1, b^(0,+)AT, CAT-1 were significantly reduced in quercetin +PEDV group(P<0.05). The study indicates that quercetin may inhibit the proliferation of PEDV in piglets by regulating intestinal amino acid transport and metabolism.