奈米矽片作為黴菌伏馬毒素B1吸附劑對雞隻肝臟蛋白質表現之影響

黴菌毒素係飼料穀物中真菌產生的有毒次級代謝產物，伏馬鐮孢毒素B1 (fumonisin B1, FB1)為Fusarium moniliforme產生之黴菌毒素，FB1汙染穀物，威脅畜禽的健康並造成經濟損失。矽鋁酸(aluminosilicates)鹽類常被作為黴菌毒素吸附劑，添加於飼料中以減少FB1吸收，為業界使用且符合成本效益的方法之一。本研究之目的為探討由蒙脫石(montmorillonite)脫層(exfoliated)製成之奈米矽片(nano silicate platelets, NSP)，作為FB1吸附劑對雞隻肝臟蛋白質表現之影響，以作為評估NSP生物毒性之基礎。試驗使用12隻一日齡雄性白肉雞，接受四種不同飼糧及飲水處理(n=3)，分別為對照組、FB1添加處理組、NSP水溶液處理組及FB1添加NSP水溶液處理組。雞隻於處理42日後犧牲並採集肝臟組織供蛋白質分析。每個肝臟樣品分別取800 μg總蛋白質進行二維電泳分析，電泳膠片上之蛋白質點以二維電泳影像分析軟體進行定量。結果顯示，在所有定量的679個肝臟蛋白質點中，有68個蛋白質點表現量具顯著的處理間差異(P<0.05)，經蛋白質胜肽指紋分析可成功鑑定63個蛋白質點之身份，此等蛋白質點分屬45個不同蛋白質。生物資訊分析結果顯示，在FB1處理組之差異表現蛋白質主要參與細胞構成(33%)、代謝過程(33%)與緊迫反應(17%)等生物過程：這些差異表現蛋白質大部份位於細胞核(33%)且與催化活性(67%)及蛋白質結合(16%)等分子功能有關，其中triosephosphate isomerase參與糖解作用。在NSP處理組中，差異表現蛋白質主要參與代謝過程(61%)，大部份位於細胞質(54%)且與催化活性(24%)、蛋白質結合(24%)及離子結合(20%)等分子功能有關，其中ATP synthase subunit beta、pyruvate carboxylase、fructose-bisphosphate aldolase C及malate dehydrogenase參與代謝過程，peroxiredoxin-1參與氧化磷酸化過程；肝臟解毒作用的關鍵酵素glutathione S-transferase及參與代謝過程的pyruvate dehydrogenase E1 component subunit beta則在FB1處理組及NSP處理組皆有顯著差異表現。由本研究結果可知，FB1可能影響與肝癌相關蛋白質之表現量，並可能促進肝臟解毒作用及影響肝細胞之代謝。NSP會改變雞隻肝臟蛋白質之表現，可能造成肝細胞氧化緊迫，而肝細胞可能藉由調控糖質新生、糖解作用與TCA cycle等路徑改變能量代謝，以應付此氧化緊迫。但當NSP與FB1結合時則會同時減低對肝細胞蛋白質表現的負面影響。Mycotoxins are a group of secondary fungal metabolites that are toxic to animals and thereby tend to cause significant financial losses in the industry. Mycotoxin fumonisin B1 (FB1) is a metabolites produced by Fusarium moniliforme. Dietary supplementation of aluminosilicates is a practical and effectively method in the industry to reduce absorption and ameliorate the toxicity of mycotoxins. This study was aimed to investigate the effect of nano silicate platelets (NSP) exfoliated from montmorillonites as a FB1 binder on the protein expression profile in the liver of broilers. Results of the study may further be applied to assess biosafety of NSP. One-day-old male broilers were assigned to the following four dietary treatments: control feed and water without NSP, feed containing 40 ppm of FB1 and water without NSP, control feed and water containing 40 ppm of NSP, and feed containing 40 ppm FB1 and water with 40 ppm NSP solution. Three birds from each group were sacrificed after 42 days of treatment and the liver samples were collected for proteomic analysis. A total of 800 μg proteins were subjected to two-dimensional gel electrophoresis and image analysis. Results showed 68 out of 679 quantified protein spots differing significantly among treatments (P