千金子制霜前后天然多元微乳对IEC-6细胞线粒体自噬性死亡的影响OA
Effects of natural microemulsion from Euphorbia Semen and Euphorbia Semen Pulveratum on mitochondrial autophagic death in IEC-6 cells
目的 探讨千金子制霜前后天然多元微乳对大鼠小肠隐窝上皮细胞(IEC-6)线粒体自噬性死亡的影响.方法 将 IEC-6 细胞分为对照组、千金子天然多元微乳组(50,100,200,400,800 μg/mL)、千金子霜天然多元微乳组(50,100,200,400,800 μg/mL),MDC 染色法观察 IEC-6 细胞自噬小体形成情况,Western blot 法检测 IEC-6 细胞中线粒体融合蛋白2(Mfn2)、视神经萎缩蛋白1(OPA1)、线粒体动力相关蛋白1(Drp1)、微管相关蛋白1 轻链 3-Ⅱ/Ⅰ(LC3-Ⅱ/Ⅰ)、螯合体1(p62)蛋白表达情况;另将 IEC-6 细胞分为对照组、3-MA 组、千金子天然多元微乳组(50,100,200,400,800 μg/mL)、3-MA 联合千金子天然多元微乳组(10 μmol/L 3-MA 预处理后加入相应浓度千金子天然多元微乳)、千金子霜天然多元微乳组(50,100,200,400,800 μg/mL)、3-MA 联合千金子霜天然多元微乳组(10 μmol/L 3-MA 预处理后加入相应浓度千金子霜天然多元微乳),MTT 法检测 IEC-6 细胞存活率.结果 与对照组比较,千金子天然多元微乳各浓度组IEC-6 细胞内自噬小体数量明显增加,绿色相对荧光强度均明显升高(P 均<0.05),细胞中 Mfn2、OPA1、p62 蛋白相对表达量均明显降低(P 均<0.05),细胞中 Drp1、LC3-Ⅱ/Ⅰ(除 50 μg/mL 浓度)蛋白相对表达量均明显升高(P 均<0.05).与千金子天然多元微乳各浓度组比较,千金子霜天然多元微乳各浓度组 IEC-6 细胞内自噬小体数量明显减少,绿色相对荧光强度均明显降低(P均<0.05),细胞中Mfn2、OPA1、p62 蛋白相对表达量均明显升高(P 均<0.05),细胞中 Drp1、LC3-Ⅱ/Ⅰ(除50 μg/mL 浓度)蛋白相对表达量均明显降低(P 均<0.05).3-MA 联合千金子天然多元微乳各浓度组IEC-6 细胞存活率均明显高于千金子天然多元微乳相应浓度组(P 均<0.05),3-MA 联合千金子霜天然多元微乳各浓度组 IEC-6 细胞存活率均明显高于千金子霜天然多元微乳相应浓度组(P 均<0.05).结论 千金子霜天然多元微乳诱导细胞线粒体融合/分裂失衡和自噬性细胞死亡的作用明显减弱,可以有效减轻千金子天然多元微乳对 IEC-6 细胞的毒性作用.
Objective It is to explore the effects of natural microemulsion from Euphorbia Semen and Euphorbia Semen Pulveratum on mitochondrial autophagic death in rat intestinal epithelioid cell line No.6(IEC-6).Methods IEC-6 cells were divided into control group,Euphorbia Semen natural microemulsion group(50,100,200,400 and 800 μg/mL)and Euphorbia Semen Pulveratum natural microemulsion group(50,100,200,400 and 800 μg/mL),the formation of autoph-agic vesicles in IEC-6 cells was observed by MDC staining,the protein expressions of mitofusin 2(Mfn2),optic atrophy 1(OPA1),dynamin-related protein1(Drp1),microtubule-associated protein1 light chain 3-Ⅱ/Ⅰ(LC3-Ⅱ/Ⅰ)and se-questosome 1(p62)in the IEC-6 cells were determined by Western blot.In addition,IEC-6 cells were divided into the follow-ing groups:control group,3-MA group,Euphorbia Semen natural microemulsion group(50,100,200,400 and 800 μg/mL),3-MA+Euphorbia Semen natural microemulsion group(treated with respective dose of Euphorbia Semen natural microemul-sion after pretreatment with 10 μmol/L 3-MA),Euphorbia Semen Pulveratum natural microemulsion group(50,100,200,400,and 800 μg/mL),and 3-MA+Euphorbia Semen Pulveratum natural microemulsion group(treated with respective dose of Euphorbia Semen Pulveratum natural microemulsion after pretreatment with 10 μmol/L 3-MA),the IEC-6 cell via-bility in each group was detected by MTT.Results Compared with the control group,the number of autophagosomes and the relative intensity of green fluorescence in IEC-6 cells were significantly increased in all concentration groups of Euphorbia Se-men natural microemulsion(all P<0.05),the relative protein expressions of Mfn2,OPA1 and p62 in the cells were all sig-nificantly reduced(all P<0.05),while the relative protein expressions of Drp1,LC3-Ⅱ/Ⅰ(except for the 50 μg/mL concentration)were all significantly increased(all P<0.05).Compared with the corresponding concentration groups of Euphorbia Semen natural microemulsion,the number of autophagosomes in IEC-6 cells of the corresponding concentration groups of Euphorbia Semen Pulveratum natural microemulsion was significantly decreased,and the relative intensity of green fluorescence was significantly decreased(all P<0.05),the relative protein expressions of Mfn2,OPA1 and p62 in the cells were all significantly increased(all P<0.05),while the relative protein expressions of Drp1,LC3-Ⅱ/Ⅰ(except for the 50 μg/mL concentration)were all significantly decreased(all P<0.05).The IEC-6 cell survival rates in all corre-sponding concentration groups of 3-MA+Euphorbia Semen natural microemulsion were significantly higher than those in the corresponding concentration groups of Euphorbia Semen natural microemulsion(all P<0.05).Similarly,the IEC-6 cell survival rates in all corresponding concentration groups of 3-MA+Euphorbia Semen Pulveratum natural microemulsion were significantly higher than those in the corresponding concentration groups of Euphorbia Semen Pulveratum natural microemul-sion(all P<0.05).Conclusion Euphorbia Semen Pulveratum natural microemulsion induced cellular mitochondrial fu-sion/disintegration imbalance and autophagic cell death were significantly weaker than those induced by Euphorbia Semen natural microemulsion,which could effectively attenuate the cytotoxic effects of Euphorbia Semen natural microemulsion on IEC-6 cells.
王慧楠;王英姿;朱海婷;张俊丽;王晓宇;陈文静;毛易凡;姜明瑞;张新宁;徐鹏
北京中医药大学中药学院,北京 102488北京中医药大学中药学院,北京 102488北京中医药大学中药学院,北京 102488北京中医药大学中药学院,北京 102488北京中医药大学中药学院,北京 102488北京中医药大学中药学院,北京 102488北京中医药大学中药学院,北京 102488北京中医药大学中药学院,北京 102488北京中医药大学中药学院,北京 102488北京中医药大学中药学院,北京 102488
医药卫生
千金子千金子霜天然多元微乳大鼠小肠隐窝上皮细胞线粒体自噬性死亡
Euphorbiae SemenEuphorbia Semen Pulveratumnatural microemulsionrat intestinal epithelioid cellmitochondrial autophagic death
《现代中西医结合杂志》 2026 (15)
2045-2050,2056,7
北京市科技新星计划项目(20240484548)中央高校基本科研业务费专项基金项目(2024-JYB-JBZD-047)国家自然科学基金资助项目(82374040)国家中医药管理局高水平建设学科项目(zyyzdxk-2023272)
评论