葛根素对饲喂氧化大豆油肉鸡小肠黏膜屏障功能及抗氧化能力的影响

Effects of Puerarin on Intestinal Barrier Function and Antioxidant Capacity of Broilers Fed with Oxidized Soybean Oil

本试验旨在研究葛根素对饲喂氧化大豆油饲粮黄羽肉鸡小肠黏膜形态结构、紧密连接蛋白基因表达及抗氧化能力的影响。试验采用2×2因子设计,因子包括油脂质量(新鲜大豆油和氧化大豆油)和葛根素添加水平(0、750 mg·kg^(-1))。选取健康1日龄雌性黄羽肉鸡360只,随机分成4个处理组,分别为新鲜大豆油饲粮组、新鲜大豆油+葛根素(750 mg·kg^(-1))饲粮组、氧化大豆油饲粮组和氧化大豆油+葛根素(750 mg·kg^(-1))饲粮组,每组6个重复,每个重复15羽。在28和56日龄时,每个重复随机选取1只鸡,取十二指肠、空肠、回肠检测小肠形态结构和测定小肠黏膜紧密连接蛋白基因表达及抗氧化指标。结果表明:1)饲喂氧化大豆油显著降低28日龄肉鸡十二指肠的绒毛高度和绒毛高度/隐窝深度(V/C)、回肠V/C和claudin-1 mRNA表达量以及56日龄肉鸡3个肠段的绒毛高度及V/C(P<0.05),显著升高56日龄3个肠段的隐窝深度(P<0.05)。饲粮中添加葛根素显著提高28日龄肉鸡空肠绒毛高度和V/C、回肠闭合小环蛋白1(ZO-1)mRNA表达量和56日龄肉鸡十二指肠V/C、空肠和回肠的绒毛高度及V/C(P<0.05),显著降低3个肠段的隐窝深度(P<0.05)。2)饲喂氧化大豆油显著升高28日龄十二指肠还原型谷胱甘肽(GSH)含量和空肠还原型谷胱甘肽过氧化物酶(GSH-Px)活性(P<0.05),并显著降低28日龄回肠GSH含量、超氧化物歧化酶(SOD)活力和总抗氧化能力(T-AOC)(P<0.05)。饲粮中添加葛根素显著提高28日龄肉鸡十二指肠GSH-Px活性(P<0.05)。饲喂氧化大豆油显著降低56日龄回肠SOD活性和T-AOC(P<0.05),饲粮中添加葛根素显著提高56日龄肉鸡十二指肠SOD和回肠SOD、GSH-Px活性(P<0.05),显著降低56日龄肉鸡空肠GSH含量、SOD活性、T-AOC和回肠T-AOC(P<0.05)。综上所述,饲喂氧化大豆油破坏肠道黏膜形态结构、降低紧密连接蛋白基因的表达量和抗氧化能力,添加葛根素可提高肉鸡小肠黏膜紧密连接蛋白基因的表达量,改善氧化损伤条件下肠道黏膜形态结构,提高抗氧化酶的活性而提高其抗氧化能力。

The experiment was conducted to investigate the effects of puerarin on mucosal morphology,mRNA expression of tight junction protein and antioxidant capacity of small intestine in yellow-feathered broilers fed oxidized soybean oil.The experiment was designed with 2×2 factors including oil quality(fresh soybean oil and oxidized soybean oil)and puerarin addition levels(0,750 mg·kg^(-1)).A total of 3601-day-old healthy female yellow-feathered broilers were randomly divided into 4 treatment groups with 6 replicates per group and 15 chickens per replicate,which were fresh soybean oil diet group,fresh soybean oil+puerarin(750 mg·kg^(-1))diet group,oxidized soybean oil diet group and oxidized soybean oil+puerarin(750 mg·kg^(-1))diet group,respectively.At 28 and 56 days of age,one chicken per replicate was randomly selected,and the morphology of small intestinal mucosa,the mRNA expression of tight junction protein and antioxidant indices of small intestine mucosa were determined.The results showed that:1)Consumption of oxidized soybean oil significantly decreased villus height and villus height/crypt depth(V/C)in duodenum of 28-day-old broilers,V/C and mRNA expression levels of claudin-1 in ileum,villus height and V/C in three intestinal segments of 56-day-old broilers(P<0.05).Consumption of oxidized soybean oil significantly increased crypt depth of three intestinal segments at 56 days of age(P<0.05).Dietary supplementation of puerarin significantly increased villus height and V/C in jejunum and Zonula occluden1(ZO-1)mRNA expression level in ileum of broilers at 28 days of age,and V/C in duodenum,jejunum and ileum and villus height in jejunum and ileum of broilers at 56 days of age(P<0.05),and significantly decreased crypt depth in three intestinal segments(P<0.05).2)Oxidized soybean oil significantly increased the glutathione(GSH)content in duodenum and activity of glutathion peroxidase(GSH-Px)in jejunum at 28 days of age(P<0.05),and significantly decreased the GSH content,superoxide dismutase(SOD)activity and total antioxidant capacity(T-AOC)in ileum at 28 days of age(P<0.05).Dietary supplementation of puerarin significantly increased the activity of GSH-Px in duodenum of broilers at 28 days of age(P<0.05).Dietary oxidized soybean oil significantly decreased the activity of SOD and T-AOC in ileum at 56 days of age(P<0.05),dietary supplementation of puerarin significantly increased the activities of SOD in duodenum and ileum,and GSH-Px in ileum(P<0.05),and significantly decreased the content of GSH and the activity of SOD,T-AOC in jejunum and T-AOC in ileum(P<0.05)of broilers at 56 days of age.In conclusion,consumption of oxidized soybean oil damaged intestinal mucosal morphology and decreased the mRNA expression of tight junction protein and antioxidant capacity of small intestinal.Puerarin could increase the mRNA expression of tight junction protein in small intestinal mucosa of broilers,improve the morphological structure of intestinal mucosa under oxidative injury,increase the activity of antioxidant enzymes and enhance the antioxidant capacity of broilers.