Gilani S, Howarth G S, Kitessa S M, Tran C D, Forder R E A, Hughes R J
1School of Animal and Veterinary Sciences,University of Adelaide,Roseworthy Campus,Adelaide, SA 5371,Australia.
3PPPI Nutrition Research Laboratory,South Australian Research & Development Institute,Roseworthy, SA 5371,Australia.
Animal. 2017 Jul;11(7):1174-1179. doi: 10.1017/S1751731116002470. Epub 2016 Nov 24.
Increased intestinal permeability (IP) can lead to compromised health. Limited in vivo IP research has been conducted in chickens. The objectives of the current study were to develop a model of increased IP utilizing lipopolysaccharide (LPS Escherichia coli O55:B5) and to evaluate IP changes using the lactulose, mannitol and rhamnose (LMR) sugar permeability test. In addition, fluorescein isothiocyanate dextran (FITC-d), d-lactate, zonula occludens (ZO-1) and diamine oxidase (DAO) permeability tests were employed. Male Ross chickens were reared until day 14 on the floor in an animal care facility and then transferred to individual cages in three separate experiments. In each of experiments 1 and 2, 36 chicks were randomly allocated to receive either saline (control) or LPS (n=18/group). Lactulose, mannitol and rhamnose sugar concentration in blood was measured at 0, 30, 60, 90, 120 and 180 min in experiment 1, at 60, 90 and 120 min in experiment 2 and at 90 min in experiment 3 (n=16/group). Lipopolysaccharide was injected intraperitoneally at doses of 0.5, 1 and 1 mg/kg BW in experiments 1, 2 and 3, respectively, on days 16, 18 and 20, whereas control received sterile saline. On day 21, only birds in experiments 1 and 2 were fasted for 19.5 h. Chicks were orally gavaged with the LMR sugars (0.25 gL, 0.05 gM, 0.05 gR/bird) followed by blood collection (from the brachial vein) as per time point for each experiment. Only in experiment 3, were birds given an additional oral gavage of FITC-d (2.2 mg/ml per bird) 60 min after the first gavage. Plasma d-lactate, ZO-1 and DAO concentrations were also determined by ELISA in experiment 3 (n=10). Administration of LPS did not affect IP as measured by the LMR sugar test compared with control. This was also confirmed by FITC-d and DAO levels in experiment 3 (P>0.05). The plasma levels of d-lactate were decreased (P<0.05). Plasma levels of ZO-1 were increased in the third experiment only and did not change in the first two experiments. Lipopolysaccharide at doses of 0.5 and 1 mg/kg did not increase IP in this model system. In conclusion, the LMR sugar can be detected in blood 90 min after the oral gavage. Further studies are needed for the applicability of LMR sugars tests.
肠道通透性增加(IP)会损害健康。在鸡身上进行的体内IP研究有限。本研究的目的是利用脂多糖(LPS,大肠杆菌O55:B5)建立IP增加的模型,并使用乳果糖、甘露醇和鼠李糖(LMR)糖通透性试验评估IP变化。此外,还采用了异硫氰酸荧光素葡聚糖(FITC-d)、d-乳酸、紧密连接蛋白(ZO-1)和二胺氧化酶(DAO)通透性试验。雄性罗斯鸡在动物饲养设施的地面饲养至14日龄,然后在三个独立实验中转至个体笼中。在实验1和实验2中,每组36只雏鸡被随机分配接受生理盐水(对照)或LPS(每组n = 18)。在实验1中,于0、30、60、90、120和180分钟测量血液中乳果糖、甘露醇和鼠李糖的浓度;在实验2中,于60、90和120分钟测量;在实验3中,于90分钟测量(每组n = 16)。在实验1、2和3中,分别于第16、18和20天腹腔注射脂多糖,剂量分别为0.5、1和1 mg/kg体重,而对照组接受无菌生理盐水。在第21天,仅实验1和实验2中的鸡禁食19.5小时。雏鸡经口灌胃LMR糖(每只鸡0.25 g乳果糖、0.05 g甘露醇、0.05 g鼠李糖),然后根据每个实验的时间点从臂静脉采血。仅在实验3中,鸡在首次灌胃60分钟后额外经口灌胃FITC-d(每只鸡2.2 mg/ml)。在实验3中(n = 10),还通过ELISA测定血浆d-乳酸、ZO-1和DAO浓度。与对照组相比,LPS给药后通过LMR糖试验测得的IP未受影响。实验3中的FITC-d和DAO水平也证实了这一点(P>0.05)。d-乳酸的血浆水平降低(P<0.05)。仅在第三个实验中血浆ZO-1水平升高,在前两个实验中未发生变化。在该模型系统中,0.5和1 mg/kg剂量的脂多糖未增加IP。总之,经口灌胃90分钟后可在血液中检测到LMR糖。LMR糖试验的适用性还需要进一步研究。