【摘要】：背景：熱應(yīng)激是誘導(dǎo)動物機體產(chǎn)生氧化應(yīng)激的重要因素之一,多酚的特殊結(jié)構(gòu)使其具有較強的抗氧化性和清除自由基能力。本實驗室的前期生化研究發(fā)現(xiàn)板栗總苞多酚(PICB)具有很強的清除自由基和抗氧化能力,在高溫?zé)釕?yīng)激情況下PICB上調(diào)AA肉雞空腸粘膜促生長、抗氧化相關(guān)基因mRNA表達。目的：分別從體外細胞試驗和體內(nèi)試驗兩個方面研究板栗總苞多酚對IEC-6細胞的促生長、抗氧化作用和對熱應(yīng)激艾拔益加(AA)肉雞血清及空腸黏膜代謝組學(xué)的影響。闡明板栗總苞多酚的促生長、抗氧化作用機制,為板栗總苞多酚作為天然植物飼料添加劑的應(yīng)用奠定理論基礎(chǔ)。方法：1、利用MTT法檢測不同濃度板栗總苞多酚對IEC-6細胞增殖活力的影響并確定最佳添加量：2、利用熒光定量PCR和Western Blottin g蛋白免疫印跡技術(shù)檢測板栗總苞多酚對IEC-6細胞(Ⅰ為常溫對照組、Ⅱ為常溫板栗總苞多酚組、Ⅲ高溫對照組、Ⅳ為高溫板栗總苞多酚組)抗氧化、促生長相關(guān)因子nRNA轉(zhuǎn)錄和蛋白表達；3、利用氣相色譜-飛行時間質(zhì)譜法分析研究板栗總苞多酚對熱應(yīng)激AA肉雞空腸黏膜和血清中小分子代謝物差異變化的影響。結(jié)果：1、常溫情況下各濃度板栗總苞多酚作用于IEC-6細胞,不同濃度細胞增殖活力與對照組相比無顯著差異；高溫情況下對照組顯著低于多酚組(P0.05),且在濃度為180μg/ml時,多酚組細胞OD值達到最大(P0.05)并與常溫對照組差異不顯著。2、常溫條件下,Ⅰ組HSP70和EGF mRNA表達量與Ⅱ組相比較無顯著變化(P0.05)；Ⅱ組SOD、EGFR和ODC mRNA表達量顯著高于Ⅰ組(P0.05)；Ⅱ組SOD蛋白表達量顯著高于Ⅰ組(P0.05),Ⅰ組和Ⅱ組EGFR和ODC蛋白表達量基本相同(P0.05)；高溫?zé)釕?yīng)激狀態(tài)下,Ⅳ組HSP70、ODC、EGF、EGFR、SOD mRNA表達量均顯著高于Ⅲ組(P0.05)；Ⅳ組SOD、ODC和EGFR蛋白表達量均高于Ⅲ組。3、與對照組相比0.3%板栗總苞多酚組顯著降低了AA肉雞血清中谷氨酸含量(P0.05),并顯著降低空腸黏膜中肌醇和蘇糖醇含量(P0.05),同時顯著提高了鳥氨酸、絲氨酸和軟脂酸含量(P0.05)。結(jié)論：板栗總苞多酚能夠顯著提高熱應(yīng)激IEC-6細胞的生長性能和抗氧化性能,緩解高溫對IEC-6細胞造成的損傷；飼料中添加0.3%板栗總苞多酚通過影響AA肉雞的谷胱甘肽代謝、碳水化合物代謝、脂肪代謝以及蛋白質(zhì)代謝途徑,提高AA肉雞的抗氧化性能和促生長作用,緩解高溫對AA肉雞造成危害。
[Abstract]:Background: heat stress is one of the important factors that induce oxidative stress in animals. The special structure of polyphenols makes it have strong antioxidant and scavenging ability of free radicals. It was found that Chestnut involucre polyphenol (PICB) had strong free radical scavenging and antioxidant activity. Under high temperature heat stress, PICB up-regulated the growth of jejunal mucosa and the expression of antioxidant related gene mRNA in the jejunum of AA broilers. Objective: to study the effects of Chestnut involucre polyphenols on the growth of IEC-6 cells in vitro and in vivo, and on the metabolism of serum and jejunum mucosa of (AA) broilers under heat stress. The mechanism of promoting growth and antioxidation of Chestnut involucre polyphenols was expounded, which laid a theoretical foundation for the application of Chestnut involucre polyphenols as natural plant feed additives. Methods: 1. The effect of different concentrations of Chestnut involucre polyphenol on the proliferation of IEC-6 cells was detected by MTT assay and the optimum dosage was determined as follows: 2. Fluorescence quantitative PCR and Western Blottin G Western blot were used to detect the antioxidation of Chestnut involucre polyphenols to IEC-6 cells (鈪，

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