当归不同炮制品多糖干预头孢噻呋钠联合LPS致鸡肝损伤效果与机制探讨

Chicken colibacillosis, one of the most serious diseases of chicken. After it is treated with antimicrobial drug, LPS will appear. The chicken liver injure induced with antimicrobial drug and LPS is common and serious. So it has great significance to research the medicine or additive for protecting liver. In prior research, We found that the polysaccharides from different processed products of Angelica sinensis have significant protecting effect on liver injury caused by carbon tetrachloride. The main mechanisms of protecting liver are mainly about intervening lipid metabolism, amino acid metabolism, and energy metabolism. Whether the polysaccharides from different processed products of Angelica Sinensis have significant protecting effect on liver injury caused by Ceftiofur Sodium and LPS in veterinary clinic? Which protecting effect is the best? And what is the protecting effect mechanism? We are not clear about. For further revealing these questions, the following studies will be carried out: For imitating the chicken liver injure model after Chicken colibacillosis is treated with Ceftiofur Sodium in veterinary clinic, to establish the chicken liver injure model caused by Ceftiofur Sodium and LPS, to measure the liver related physiological and biochemical index, anti-oxidation index, immunity index, combining the means of metabolomics and proteomics, to research the variation of liver protein spectrum and metabolites in serum and liver between the liver injure model and the intervention of polysaccharides from different processed products of Angelica sinensis on liver injury model, screen the target proteins and metabolites of protecting liver injure of polysaccharides from different processed products of Angelica sinensis, and discusses the mechanisms of protecting liver injure caused by Ceftiofur Sodium and LPS of polysaccharides from different processed products of Angelica sinensis.

鸡大肠杆菌病，危害养鸡业最严重疾病之一。采用抗生素治疗会产生内毒素（LPS），两者协同引起鸡肝损伤普遍存在且危害严重，因此研发保肝药物或添加剂意义重大。前期研究我们发现当归不同炮制品多糖对四氯化碳致肝损伤有较好保护作用，其机制与干预脂质代谢、氨基酸代谢与能量代谢有关。而当归不同炮制品多糖对头孢噻呋钠联合LPS致鸡肝损伤是否具有保肝效果，哪种炮制品多糖效果最佳，其机制如何都不清楚。为揭示这些问题，本研究拟模仿头孢噻呋钠干预鸡大肠杆菌后所引发的肝损伤，建立头孢噻呋钠联合LPS致鸡肝损伤模型，检测肝脏相关生理生化指标、抗氧化指标和免疫调节指标，结合代谢组学与蛋白组学手段，研究肝损伤模型和当归不同炮制品多糖对鸡肝损伤的肝脏差异性蛋白质谱及血清和肝脏的内源性代谢物变化，筛选当归不同炮制品多糖发挥保肝作用的蛋白质和代谢物，探讨当归不同炮制品多糖干预头孢噻呋钠联合LPS所致肝损伤机制。

鸡大肠杆菌病，危害养鸡业最严重疾病之一。采用抗生素治疗会产生细菌内毒素（脂多糖，LPS），两者协同引起鸡肝损伤危害严重。本课题主要研究内容：（1）头孢噻呋钠联合LPS致鸡肝损伤模型建立；（2）当归不同炮制品多糖对头孢噻呋钠联合LPS致鸡肝损伤干预效果研究；（3）分别从肝损伤、抗氧化、免疫炎症因子、病理组织学观察、代谢层面、蛋白层面阐明头孢噻呋钠联合LPS致鸡肝损伤发生机制及当归不同炮制品（炭当归、生当归）多糖的干预机制。结果表明：（1）给鸡皮下注射头孢噻呋钠（5 mg/kg），1次/d，连续3 d，第3天皮下注射完头孢噻呋钠后注射LPS（4 mL/kg）可复制鸡肝损伤模型；（2）证实当归不同炮制品多糖对头孢噻呋钠联合LPS致鸡肝损伤有保护效果，筛选出炭当归多糖可作为保肝药物；（3）头孢噻呋钠联合LPS造模后，产生氧化应激、炎症反应，严重影响鸡体内的跨膜转运、氨基酸代谢、亚油酸代谢、花生四烯酸代谢、mTOR信号通路等重要代谢通路，引发肝损伤，涉及10个重要代谢物与7个关键蛋白；（4）当归不同炮制品多糖保肝效果具有差异，炭当归多糖防治效果优于生当归多糖。炭当归多糖防治头孢噻呋钠联合LPS致鸡肝损伤的机制可能是通过恢复跨膜转运、氨基酸代谢、mTOR信号通路、花生四烯酸代谢、亚油酸代谢等通路紊乱，回调抗氧化、炎症相关酶与细胞因子表达而调节抗氧化能力与免疫反应，促进肝组织损伤修复，涉及9个重要差异代谢物和7个关键蛋白。生当归多糖防治头孢噻呋钠联合LPS致鸡肝损伤的机制主要涉及恢复跨膜转运、氨基酸代谢等代谢通路紊乱，部分回调免疫炎症细胞因子，促进肝组织损伤修复，涉及4个差异代谢物。本研究证实头孢噻呋钠与LPS共存可引发鸡肝损伤，炭当归多糖筛选与机制研究为研发新型兽用保肝药奠定基础，对绿色健康畜禽养殖业中减抗替抗、保护动物福利、保障动物性食品安全均具有重要意义，具有较好的应用前景。