本文選題：猴頭菇多糖 + 過氧化氫　； 參考：《畜牧獸醫(yī)學報》2017年09期

【摘要】：旨在探究在H_2O_2誘導的氧化應激損傷狀態(tài)下,猴頭菇多糖(HEPs)對IPEC-J2細胞的保護作用。將IPEC-J2細胞分為5組:空白組、模型組、HEPs低劑量組、HEPs中劑量組、HEPs高劑量組。采用MTT法分別選取試驗用的H_2O_2和HEPs的最佳劑量;利用DCFH-DA染色觀察并測定細胞內(nèi)活性氧簇(ROS)的含量;通過比色法檢測培養(yǎng)細胞上清液中丙二醛(MDA)的含量,細胞中超氧化物歧化酶(SOD)和過氧化氫酶(CAT)的活力以及乳酸脫氫酶(LDH)的釋放率;應用實時熒光定量PCR(qRT-PCR)檢測緊密連接蛋白ZO-1基因的轉錄水平。結果顯示,(1)構建IPEC-J2細胞氧化應激模型的H_2O_2濃度為0.4mmol·L-1。(2)H_2O_2能夠極顯著地增加細胞內(nèi)的ROS含量,誘導細胞凋亡,同時極顯著地降低了SOD和CAT的活力,極顯著地升高MDA含量與LDH釋放率。但是,與模型組相比,經(jīng)過HEPs預處理后,細胞內(nèi)ROS的含量極顯著地降低(P0.01);同時極顯著地提高了SOD和CAT的活力(P0.01),極顯著地降低了MDA含量與LDH釋放率(P0.01)。(3)實時熒光定量PCR結果顯示,H_2O_2能夠極顯著地降低ZO-1基因的轉錄量(P0.01),與模型組比較,HEPs能夠極顯著的提高ZO-1基因的轉錄量(P0.01);Western blot結果顯示ZO-1蛋白的表達量與ZO-1基因類似。可見,猴頭菇多糖對H_2O_2誘導氧化損傷狀態(tài)下的IPEC-J2細胞具有保護作用。
[Abstract]:To investigate the protective effect of hericium Erinaceus polysaccharides (Hep) on IPEC-J2 cells under oxidative stress induced by H_2O_2.IPEC-J2 cells were divided into five groups: blank group, model group, low dose group, middle dose group and high dose group.MTT method was used to select the best dosage of H_2O_2 and HEPs, DCFH-DA staining was used to observe and measure the content of reactive oxygen species (Ros) in cells, and the content of malondialdehyde (MDA) in supernatant of cultured cells was detected by colorimetric method.The activity of superoxide dismutase (SOD) and catalase (catalase) and the release rate of lactate dehydrogenase (LDH) were determined by real-time fluorescence quantitative analysis of ZO-1 gene transcription level.The results showed that the H_2O_2 concentration of 0.4mmol L-1.(2)H_2O_2 in the oxidative stress model of IPEC-J2 cells was 0.4mmol L-1.(2)H_2O_2, which could significantly increase the ROS content, induce apoptosis, decrease the activity of SOD and CAT, and significantly increase the content of MDA and the release rate of LDH.However, compared with the model group, after HEPs pretreatment,The results of real-time fluorescence quantitative PCR analysis showed that H2O-2 could significantly decrease the activity of ZO-1 gene, and increase the activity of SOD and CAT, and decrease the content of MDA and the release rate of LDH.Compared with the model group, Hep could significantly increase the transcription quantity of ZO-1 gene. The results of Western blot showed that the expression of ZO-1 protein was similar to that of ZO-1 gene.Therefore, hericium Erinaceus polysaccharides can protect IPEC-J2 cells from oxidative damage induced by H_2O_2.
【作者單位】： 福建農(nóng)林大學福建省獸醫(yī)中藥與動物保健重點實驗室;中西獸醫(yī)結合與動物保健福建省高等學校重點實驗室;
【基金】：國家自然科學基金(31502110)
【分類號】：S853.7

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