【摘要】 为探究饲料添加剂麻叶荨麻(Urtica cannabina L.,UCL)对雏鸡的抗炎效果，试验将64只3日龄三黄鸡(雌雄各半)随机分为8组，分别为对照组(CO组)、模型组[脂多糖(LPS)组]、1%UCL组、1%UCL+LPS组、3%UCL组、3%UCL+LPS组、9%UCL组、9%UCL+LPS组，每组8只。LPS组和UCL+LPS组肉仔鸡按体重腹腔注射750μg/kg LPS,CO组和UCL组注射生理盐水，CO组和LPS组饲喂基础日粮，UCL组和UCL+LPS组在基础日粮中分别添加相应剂量的UCL粉末。试验第22天，所有雏鸡宰杀后解剖并采集十二指肠样本进行苏木精-伊红(H.E.)染色和糖原PAS染色，使用光学显微镜拍照并用Image J 1.48v软件测量十二指肠绒毛高度(VH)、隐窝深度(CD)及杯状细胞数，并统计VH/VD。采用实时荧光定量PCR检测白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、白细胞介素-1β(IL-1β)、核转录因子(NF)-κB基因mRNA相对表达量；采用Western-blot分析IL-6、IL-8、肿瘤坏死因子-α(TNF-α)、NF-κB p65蛋白相对表达量；分析各组微生物多样性指数及在属水平上的群落组成，并进行肠道微生物群的主成分分析。结果表明：UCL富含黄酮类化合物、酚酸、氨基酸、有机酸、脂肪酸和维生素等营养物质。与CO组相比，LPS组VH、VH/VD和杯状细胞数显著降低(P<0.05),LPS组IL-6、IL-8、IL-1β和NF-κB基因mRNA相对表达量均显著增加(P<0.05);除TNF-α外，LPS组IL-6、IL-8和NF-κB p65蛋白相对表达量显著增加(P<0.05)。与LPS组相比，1%UCL+LPS组、3%UCL+LPS组和9%UCL+LPS组VH、VH/VD显著增加(P<0.05),CD显著降低(P<0.05);1%UCL+LPS组、3%UCL+LPS组杯状细胞数显著增加(P<0.05);1%UCL+LPS组、3%UCL+LPS组和9%UCL+LPS组IL-8、IL-1β基因mRNA相对表达量显著降低(P<0.05);1%UCL+LPS组、3%UCL+LPS组NF-κB基因mRNA相对表达量显著降低(P<0.05);除9%UCL组、9%UCL+LPS组外，其余各组IL-6、IL-8、NF-κB p65蛋白相对表达量均显著降低(P<0.05);9%UCL组IL-8蛋白相对表达量显著降低(P<0.05)。LPS组的Ace指数低于CO组(P>0.05),1%UCL组、3%UCL组的Ace指数极显著高于CO组(P<0.01),3%UCL+LPS组的Ace指数极显著高于LPS组(P<0.001)。与CO组相比，LPS组雏鸡肠道菌群中乳杆菌属相对丰度下降，1%UCL组、3%UCL组、3%UCL+LPS组拟杆菌属、布劳特菌属相对丰度升高；与LPS组相比，9%UCL组和9%UCL+LPS组乳杆菌属相对丰度均升高。说明在日粮中添加1%～3%UCL对LPS诱导的雏鸡十二指肠结构的损伤有恢复作用，并对LPS造成的肠道炎症因子表达的异常增加有显著抑制作用。更多还原

【Abstract】 In order to explore the anti-inflammatory effects of the feed additive hemp leaf nettle(Urtica cannabina L., UCL), 64 three-day-old Sanhuang chickens(male-to-female ratio was 1∶1) were randomly divided into 8 groups: the control group(CO), the model group(lipopolysaccharide [LPS] group), 1% UCL group, 1% UCL+LPS group, 3% UCL group, 3% UCL+LPS group, 9% UCL group, and 9% UCL+LPS group, with 8 chickens in each group. Broiler chickens in the LPS group and the UCL+LPS group were intraperitoneally injected with 750 μg/kg LPS according to their body weight, while those in the CO group and the UCL group were injected with saline. The animals in the CO group and the LPS group were fed with basal diet, while the animals in the UCL group and UCL+LPS group were given the basal diet supplemented with corresponding dose powder. On the 22nd day of the experiment, all chicks were slaughtered, and duodenal samples were collected for hematoxylin and eosin(H.E.) staining and glycogen PAS staining. Photos were taken using an optical microscope, and the duodenal villus height(VH), crypt depth(CD), and number of goblet cells were measured using Image J 1.48v software, and VH/VD was counted. The relative mRNA expression levels of interleukin-6(IL-6), interleukin-8(IL-8), interleukin-1β(IL-1β), and nuclear factor(NF)-κB genes were detected by real-time quantitative PCR. The relative protein expression levels of IL-6, IL-8, tumor necrosis factor-α(TNF-α), and NF-κB p65 were analyzed by Western-blot. The microbial diversity indices and genus-level community composition of each group were analyzed, and principal component analysis of the gut microbiota was conducted. The results showed that UCL was rich in flavonoids, phenolic acids, amino acids, organic acids, fatty acids, vitamins, and other nutrients. Compared with the CO group, the VH, VH/VD, and goblet cell numbers in the LPS group were significantly decreased(P<0.05), while the relative mRNA expression levels of IL-6, IL-8, IL-1β, and NF-κB genes were significantly increased(P<0.05). Except for TNF-α, the relative protein expression levels of IL-6, IL-8, and NF-κB p65 in the LPS group were significantly increased(P<0.05). Compared with the LPS group, VH and VH/VD in the 1% UCL+LPS group, 3% UCL+LPS group, and 9% UCL+LPS group were significantly increased(P<0.05) and CD was significantly decreased(P<0.05); the number of goblet cells was significantly increased in the 1% UCL+LPS group and 3% UCL+LPS group(P<0.05); the relative mRNA expression levels of IL-8 and IL-1β genes in the 1% UCL+LPS group, 3% UCL+LPS group, and 9% UCL+LPS group were significantly reduced(P<0.05); the relative mRNA expression level of NF-κB gene was significantly reduced in the 1% UCL+LPS group and 3% UCL+LPS group(P<0.05). Compared with the LPS group, except for the 9% UCL+LPS group, the relative expression levels of IL-6, IL-8, and NF-κB p65 proteins were significantly reduced in all other groups(P<0.05); the relative expression levels of IL-8 protein were significantly reduced in the 9%UCL group(P<0.05). The Ace index in the LPS group was lower than that in the CO group(P>0.05); the Ace index in the 1%UCL, 3%UCL groups was extremely higher than that in the CO group(P<0.001); the Ace index in the 3% UCL+LPS group was extremely significantly higher than that in the LPS group(P<0.001). Compared with the CO group, the relative abundance of Lactobacillus in the intestinal microbiota of chicks in the LPS group was decreased, while the relative abundance of Bacteroides and Blautia was relatively high in the 1% UCL group, 3% UCL group, and 3% UCL + LPS group. Compared with the LPS group, the relative abundance of Lactobacillus was increased in both the 9% UCL group and the 9% UCL+LPS group. The results indicated that adding 1%-3% UCL to the diet had a restorative effect on the damage of duodenal structural in chicks induced by LPS, and significantly inhibited the abnormal increase in the expression of intestinal inflammatory factors caused by LPS.更多还原