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作 者:范崇熙 张云水 孙兵兵 王晓莹 赵东林 杨全龙 石学汇 刘磊 FAN Chong-xi;ZHANG Yun-shui;SUN Bing-bing;WANG Xiao-ying;ZHAO Dong-lin;YANG Quan-long;SHI Xue-hui;LIU Lei(Department of Gastroenterology,Air Force Medical Center,Beijing 100142,China;Department of Critical Care Medicine,Air Force Medical Center,Beijing 100142,China)
机构地区:[1]空军特色医学中心消化内科,北京100142 [2]空军特色医学中心重症医学科,北京100142
出 处:《局解手术学杂志》2023年第10期839-845,共7页Journal of Regional Anatomy and Operative Surgery
基 金:国家自然科学基金青年基金项目(81702731);北京市海淀区卫生健康发展科研培育计划(HP2021-19-80801);空军特色医学中心博士助推项目(21ZT16);2021年空军军医大学临床研究项目(2021LC2201)。
摘 要:目的探讨紫檀芪(PTE)对过氧化氢(H_(2)O_(2))诱导HT22细胞氧化应激损伤的保护作用及调控机制。方法不同浓度的PTE处理HT22细胞12 h,并构建H_(2)O_(2)氧化应激损伤模型。CCK-8法检测细胞活力,比色法检测细胞乳酸脱氢酶(LDH)释放量,TUNEL法染色检测细胞凋亡,DCFH-DA测定活性氧(ROS)水平,JC-1标记线粒体膜电位(MMP),并通过Western blot检测内质网应激和凋亡相关蛋白表达情况。结果CCK-8结果显示,PTE浓度小于10.0μmol/L时,HT22细胞活性无明显变化。H_(2)O_(2)可显著抑制HT22细胞活性,5.0μmol/L和10.0μmol/L PTE可有效增加损伤后细胞活性,减少LDH释放量,抑制凋亡及相关蛋白的表达,同时减少细胞内ROS,稳定MMP和细胞色素C分布。Western blot检测发现,PTE降低H_(2)O_(2)损伤后PERK和eIF2α的磷酸化水平以及下调Bip、CHOP的表达。结论PTE通过维持线粒体稳态、抑制内质网应激,进而减轻氧化应激诱导的细胞凋亡,最终发挥保护HT22细胞抗H_(2)O_(2)损伤的作用。Objective To investigate the protective effect and regulatory mechanism of pterostilbene(PTE)on hydrogen peroxide(H_(2)O_(2))-induced oxidative stress injury in HT22 cells.Methods HT22 cells were treated with different concentrations of PTE for 12 hours,and the H_(2)O_(2)oxidative stress injury model was established.The cell viability was detected by CCK-8 assay.The release of lactate dehydroge⁃nase(LDH)was evaluated by colorimetry.The apoptosis was detected by TUNEL staining method.The reactive oxygen species(ROS)level was detected by DCFH-DA,and the mitochondrial membrane potential(MMP)was marked by JC-1.Western blot was used to detect the expression of endoplasmic reticulum stress and apoptosis-related proteins.Results The results of CCK-8 showed that the activity of HT22 cells was not significantly changed when the concentration of PTE was less than 10.0μmol/L.H_(2)O_(2)could significantly inhibit the activity of HT22 cells,5.0μmol/L and 10.0μmol/L PTE could effectively increase the activity of injured cells,reduce the release of LDH,inhibit the expression of apoptosis and related proteins,and reduce intracellular ROS,stabilize MMP and the distribution of cytochrome C.Western blot showed that PTE decreased the phosphorylations level of PERK and eIF2α,and down-regulated the expression of Bip and CHOP after H_(2)O_(2)injury.Conclusion PTE protects HT22 cells against H_(2)O_(2)injury through stabilizing mitochondrial homeostasis,inhibiting endoplasmic reticulum stress,and then reducing oxidative stress-induced apoptosis.
关 键 词:紫檀芪 HT22细胞 内质网应激 线粒体稳态 氧化应激
分 类 号:R741.05[医药卫生—神经病学与精神病学]
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