本文選題：猴頭菇多糖 + 過氧化氫�。� 參考：《畜牧獸醫(yī)學(xué)報(bào)》2017年09期

【摘要】：旨在探究在H_2O_2誘導(dǎo)的氧化應(yīng)激損傷狀態(tài)下,猴頭菇多糖(HEPs)對(duì)IPEC-J2細(xì)胞的保護(hù)作用。將IPEC-J2細(xì)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(yàn)用的H_2O_2和HEPs的最佳劑量;利用DCFH-DA染色觀察并測(cè)定細(xì)胞內(nèi)活性氧簇(ROS)的含量;通過比色法檢測(cè)培養(yǎng)細(xì)胞上清液中丙二醛(MDA)的含量,細(xì)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(yīng)用實(shí)時(shí)熒光定量PCR(qRT-PCR)檢測(cè)緊密連接蛋白ZO-1基因的轉(zhuǎn)錄水平。結(jié)果顯示,(1)構(gòu)建IPEC-J2細(xì)胞氧化應(yīng)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(xì)胞內(nèi)的ROS含量,誘導(dǎo)細(xì)胞凋亡,同時(shí)極顯著地降低了SOD和CAT的活力,極顯著地升高M(jìn)DA含量與LDH釋放率。但是,與模型組相比,經(jīng)過HEPs預(yù)處理后,細(xì)胞內(nèi)ROS的含量極顯著地降低(P0.01);同時(shí)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(shí)時(shí)熒光定量PCR結(jié)果顯示,H_2O_2能夠極顯著地降低ZO-1基因的轉(zhuǎn)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(zhuǎn)錄量(P0.01);Western blot結(jié)果顯示ZO-1蛋白的表達(dá)量與ZO-1基因類似�？梢�,猴頭菇多糖對(duì)H_2O_2誘導(dǎo)氧化損傷狀態(tài)下的IPEC-J2細(xì)胞具有保護(hù)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(xué)福建省獸醫(yī)中藥與動(dòng)物保健重點(diǎn)實(shí)驗(yàn)室;中西獸醫(yī)結(jié)合與動(dòng)物保健福建省高等學(xué)校重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金(31502110)
【分類號(hào)】：S853.7

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