文献类型：期刊文献

中文题名：鸡内金多糖对高糖诱导的胰岛β细胞活性和Nrf2信号通路的影响

英文题名：Effects of polysaccharides from Endothelium Corneum Gigeriae Galli on high glucose?induced isletβcell activity and Nrf2 signaling pathway

作者：丁叶蓉[1,2];周春楠[2];王苑铭[2]

第一作者：丁叶蓉

机构：[1]兰州市七里河区人民医院公卫科,甘肃兰州730050;[2]甘肃中医药大学附属医院内分泌科,甘肃兰州730200

第一机构：兰州市七里河区人民医院公卫科,甘肃兰州730050

年份：2024

卷号：58

期号：11

起止页码：90

中文期刊名：上海中医药杂志

外文期刊名：Shanghai Journal of Traditional Chinese Medicine

收录：CSTPCD;;CSCD:【CSCD_E2023_2024】；

基金：甘肃省卫健委中医药科研课题(GZKP-2020-30);兰州市科技发展计划项目(2022-ZD-89)。

语种：中文

中文关键词：糖尿病;鸡内金多糖;胰岛β细胞;核因子E2相关因子;作用机制;中药研究

外文关键词：diabetes mellitus;polysaccharides from Endothelium Corneum Gigeriae Galli(PECG);isletβcell;nuclear factor E2-related factor(Nrf2);mechanism of action;traditional Chinese herbal medicine research

摘要：目的探究鸡内金多糖对高糖诱导后胰岛β细胞相关活性和对超氧化物歧化酶(SOD)、丙二醛(MDA)以及核因子E2相关因子2(Nrf2)等相关指标所产生的影响。方法取胰岛β细胞MIN6,培养检测其活性后分为正常组(5.5 mmol·L^(-1)正常糖处理4 h)、高糖组(高脂高糖处理1 h,33.3 mmol·L^(-1)葡萄糖+0.25 mmol·L^(-1)棕榈酸)、鸡内金多糖低剂量组(20μmol·L^(-1)鸡内金多糖预处理4 h+高脂高糖处理24 h)、鸡内金多糖高剂量组(80μmol·L^(-1)鸡内金多糖预处理4 h+高脂高糖处理24 h),采用5-乙炔基-2′脱氧尿嘧啶核苷(EdU)检测各组细胞增殖率,缺口末端标记法(TUNEL)检测细胞凋亡,分析细胞周期,细胞划痕实验检测MIN6细胞迁移功能,检测MIN6细胞培养液中SOD、MDA水平,免疫印迹法检测MIN6细胞中Nrf2、醌氧化还原酶1(NQO1)和血红素单加氧酶-1(HO-1)蛋白表达,实时荧光定量逆转录聚合酶链式反应(RT-qPCR)检测MIN6细胞中Nrf2、HO?1、NQO1 mRNA表达。结果与正常组比较,高糖组MIN6细胞增殖率,SOD及Nrf2、HO-1、NQO1 mRNA和蛋白均降低,MDA及细胞凋亡率升高(P<0.05);与高糖组比较,鸡内金多糖低剂量组MIN6细胞增殖率,SOD及Nrf2、HO-1、NQO1 mRNA和蛋白升高、MDA及细胞凋亡率降低(P<0.05);与鸡内金多糖低剂量组比较,鸡内金多糖高剂量组MIN6细胞增殖率,SOD及Nrf2、HO-1、NQO1 mRNA和蛋白升高,MDA及细胞凋亡率降低(P<0.05)。与正常组比较,高糖组MIN6细胞周期G_(0)/G_(1)升高,S、G_(2)/M降低(P<0.05);与高糖组比较,鸡内金多糖低剂量组MIN6细胞周期G_(0)/G_(1)降低,S和G_(2)/M升高(P<0.05);与鸡内金多糖低剂量组比较,鸡内金多糖高剂量组MIN6细胞周期G_(0)/G_(1)降低,S、G_(2)/M升高(P<0.05)。划痕结果显示,0 h各组空白区无差异;24 h高糖组空白区增大;鸡内金多糖低剂量组空白区域有所缩小;鸡内金多糖高剂量组空白区域总体显著变窄。结论鸡内金多糖能够促进胰岛β细胞MIN6的增殖,抑制其凋亡,可能与抑制Nrf2等相关通路相关。
Objective To investigate the effects of polysaccharides from Endothelium Corneum Gigeriae Galli(PECG)on the activity of isletβcells induced by high glucose,superoxide dismutase(SOD),malondialdehyde(MDA)and nuclear factor E2 related factor 2(Nrf2).Methods Isletβcells MIN6 were cultured to detect their activity and then divided into normal group(5.5 mmol·L^(-1) normal glucose treatmentfor for 4 h)and high glucose group(high lipid and high glucose treatment for 1 h,33.3 mmol·L^(-1) glucose+0.25 mmol·L^(-1) palmitic acid),low dose of PECG(20μmol·L^(-1) PECG pretreatment for 4 h+high fat and high sugar treatment for 24 h),high dose of PECG(80μmol·L^(-1) PECG pretreatment for 4 h+high fat and high sugar treatment for 24 h),cell proliferation rate was detected by 5-acetylidene-2'deoxyuracil ribonucleoside(EdU),apoptosis was detected by notch end labeling(TUNEL),cell cycle was analyzed,and migration function of MIN6 cells was detected by cell scratch assay.The levels of SOD and MDA in MIN6 cell culture medium were detected,and the protein expressions of Nrf2,quinone oxidoreductase 1(NQO1)and heme monooxygenase-1(HO-1)in MIN6 cells were detected by Western blot.Real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-qPCR)was used to detect Nrf2,HO?1 and NQO1 mRNA expression in MIN6 cells.Results Compared with the normal group,the high-glucose group showed decreased cell proliferation of MIN6 cells,as well as reduced levels of SOD and decreased expression of Nrf2,HO-1,and NQO1 mRNA and proteins.Additionally,there were increases in MDA levels and cell apoptosis(P<0.05).Relative to the high-glucose group,the low-dose PECG group showed higher proliferation of MIN6 cells and elevated levels of SOD,as well as increased expression of Nrf2,HO-1,and NQO1 mRNA and proteins.It also showed lower MDA levels and reduced cell apoptosis(P<0.05).Compared with the low-dose PECG group,the high-dose PECG group showed increased proliferation of MIN6 cells,elevated levels of SOD,and increased expression of Nrf2,HO-1,and NQO1 mRNA and proteins.There were reductions in MDA levels and cell apoptosis(P<0.05).Compared with the normal group,the high-glucose group showed an increased G_(0)/G_(1) phase and decreased S and G_(2)/M phases in the cell cycle of MIN6 cells(P<0.05).Compared with the high-glucose group,the low-dose PECG group showed a decrease in the G_(0)/G_(1) phase and increases in the S and G_(2)/M phases(P<0.05).Compared with the low-dose PECG group,the high-dose PECG group showed further reductions in the G_(0)/G_(1) phase and increases in the S and G_(2)/M phases(P<0.05).The scratch assay results revealed no differences in the gap area at 0 h among all groups.However,at 24 h,the gap area in the high-glucose group increased,whereas it decreased in the low-dose PECG group and was significantly narrowed in the high-dose PECG group.Conclusion PECG can promote the proliferation of MIN6 isletβcells and inhibit their apoptosis,and the effect is likely associated with the suppression of the Nrf2 signaling pathway and related pathways.