桑叶多糖含药血清对体外高糖培养INS-1细胞氧化应激损伤的保护作用研究  

Protective Effects of Serum Containing Mulberry Leaf Polysaccharide on Oxidative Stress Injury in INS⁃1 Cells Cultured in High Glucose in vitro

作　　者：刘冉 刘杨阳 孙晓梅 黄淑贤 任春久[1,2] 高绘菊 张升祥[1,2] 殷培超 Liu Ran;Liu Yangyang;Sun Xiaomei;Huang Shuxian;Ren Chunjiu;Gao Huiju;Zhang Shengxiang;Yin Peichao(College of Forest,Shandong Agricultural University,Tai’an Shandong 271018,China;Key Laboratory of Silviculture in Downstream Areas of the Yellow River,National Forestry and Grassland Administration,Tai’an Shandong 271018,China;Hairun Cocoon Silk Co.,Ltd,Chuxiong Yunnan 675000,China)

机构地区：[1]山东农业大学林学院,山东泰安271018 [2]黄河下游森林培育国家林业和草原局重点实验室,山东泰安271018 [3]云南海润茧丝绸有限公司,云南楚雄675000

出　　处：《蚕业科学》2023年第4期349-358,共10页ACTA SERICOLOGICA SINICA

基　　金：山东省蚕桑产业技术体系建设项目(SDAIT-18-08);财政部和农业农村部国家现代农业产业技术体系项目;云南省院士专家工作站项目(202105AF150059)。

摘　　要：采用含桑叶多糖(mulberry leaf polysaccharide,MLP)的大鼠血清干预处理体外高糖培养的INS-1细胞,分别测定含MLP血清对高糖培养INS-1细胞中胰岛素含量、氧化应激标志物含量、细胞形态、抗氧化酶活性的影响,结果表明,MLP可明显提高高糖培养INS-1细胞中胰岛素含量,有效降低8-OHdG和丙二醛(MDA)含量,细胞形态明显改善,细胞中抗氧化酶SOD、CAT和GPx的活性明显升高,ROS含量显著降低。采用酶联免疫吸附测定MLP对高糖培养INS-1细胞线粒体呼吸链酶活性的影响,结果表明,MLP可显著提高高糖培养INS-1细胞线粒体呼吸链ComplexⅠ-Ⅳ复合酶的活性。最后采用Hoechst 33342染色、JC-1染色和免疫荧光组化法检测MLP对高糖培养INS-1细胞凋亡的影响,结果表明,MLP可明显提高线粒体膜电位,减少细胞色素C(Cyt-C)在胞浆中的表达,抑制细胞凋亡。上述结果表明,MLP可能通过提高细胞中抗氧化酶含量,改善线粒体能量代谢来减少氧化应激导致的细胞凋亡。The rat serum containing mulberry leaf polysaccharide(MLP)was used to intervene in the treatment of INS⁃1 cells cultured in high glucose in vitro,and the effects of MLP on the insulin content,oxidative stress marker content,cell morphology,and antioxidant enzyme activity of INS⁃1 cells cultured in high glucose were measured respectively.The re⁃sults showed that MLP could markedly increase the insulin content,effectively reduce the content of 8⁃OHdG and malon⁃aldehyde(MDA),significantly improve the cell morphology,increase the activity of antioxidant enzymes SOD,CAT,and GPx,and reduce the ROS content in INS⁃1 cells cultured in high glucose.Furthermore,the effects of MLP on the ac⁃tivity of mitochondrial respiratory chain enzymes of INS⁃1 cells cultured in high glucose were detected by enzyme⁃linked immunosorbent assay.The results suggested that MLP could significantly increase the activity of complexⅠ-Ⅳin mito⁃chondrial respiratory chain of INS⁃1 cells cultured in high glucose.Finally,Hoechst 33342 staining,JC⁃1 staining and im⁃munofluorescence histochemistry were used to detect the effect of MLP on the apoptosis of INS⁃1 cells cultured in high glucose.As the results displayed,MLP could significantly promote mitochondrial membrane potential,reduce the ex⁃pression of cytochrome C(Cyt⁃C)in the cytoplasm,and inhibit cell apoptosis.In all,the above results suggested that MLP may reduce cell apoptosis caused by oxidative stress by increasing the content of antioxidant enzymes in cells and improving the mitochondrial energy metabolism.

关 键 词：桑叶多糖 INS-1细胞 氧化应激 线粒体 凋亡 

分 类 号：S886.9[农业科学—特种经济动物饲养]