目的探讨金雀根和黄芪配伍对糖尿病肾病(DKD)大鼠的影响。方法将SD大鼠随机分为正常组、模型组、恩格列净组、金雀根组、黄芪组、配伍组,每组10只。采用单侧切除肾脏联合腹腔注射链脲佐菌素(STZ)复制DKD大鼠模型,造模成功后灌胃相应剂...目的探讨金雀根和黄芪配伍对糖尿病肾病(DKD)大鼠的影响。方法将SD大鼠随机分为正常组、模型组、恩格列净组、金雀根组、黄芪组、配伍组,每组10只。采用单侧切除肾脏联合腹腔注射链脲佐菌素(STZ)复制DKD大鼠模型,造模成功后灌胃相应剂量药物8周。第0、4、8周检测24 h尿微量白蛋白(24 h U-mALB);ELISA法检测Scr、BUN、CysC、MDA水平及SOD活性;荧光探针法检测肾组织ROS表达;HE、PAS、Masson、PASM-Masson染色观察肾组织病理结构改变;免疫组织化学法检测肾组织NOX4、Drp1、MFN2、P62表达;Western blot法检测肾组织PINK1、MFN2、Parkin、LC3-Ⅱ/Ⅰ、P62、p-Drp1蛋白表达。结果与模型组比较,各给药组大鼠24 h U-mALB、BUN、Scr、CysC水平均降低(P<0.01),肾组织病理性结构损伤改善;血清和组织MDA水平降低(P<0.01),SOD活性升高(P<0.01);肾组织PINK1、MFN2、Parkin、LC3-Ⅱ/Ⅰ蛋白表达升高(P<0.05,P<0.01),p-Drp1、P62蛋白表达降低(P<0.01),其中配伍组作用优于单用药组(P<0.05,P<0.01)。结论金雀根和黄芪配伍可能是通过调控PINK1/MFN2/Parkin通路改善线粒体动力学,激活线粒体自噬,抑制氧化应激,减轻肾脏病理损伤,改善DKD大鼠肾功能。展开更多
目的观察重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对缺血/再灌注损伤大鼠心肌细胞Mitofusin2(Mfn2)蛋白表达的影响及其抗心肌细胞凋亡的作用。方法选取成年SD大鼠35只,随机分为正常组(Normal),假手术组(Sham),缺...目的观察重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对缺血/再灌注损伤大鼠心肌细胞Mitofusin2(Mfn2)蛋白表达的影响及其抗心肌细胞凋亡的作用。方法选取成年SD大鼠35只,随机分为正常组(Normal),假手术组(Sham),缺血再灌注组(I/R),缺血再灌注EPO治疗组(I/R+EPO)。各组分别于再灌注3h和24h后,剪取心脏缺血/再灌注损伤区域,用脱氧核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测心肌细胞凋亡,免疫组化法检测Mfn2蛋白的表达。结果再灌注3h和24h后,与正常组和假手术组相比,I/R组Mfn2蛋白的表达和心肌细胞凋亡均显著增加;与I/R组相比,I/R+EPO组Mfn2蛋白的表达和心肌细胞凋亡均显著降低。结论EPO可以下调缺血再灌注损伤后心肌细胞Mfn2蛋白的表达,抑制心肌细胞的凋亡。展开更多
Mitofusin2 (Mfn2) plays a pivotal role in the proliferation and apoptosis of vascular smooth muscle cells (VSMCs). The purpose of this study was to investigate the effects of Mfn2 on the traffick- ing of intracell...Mitofusin2 (Mfn2) plays a pivotal role in the proliferation and apoptosis of vascular smooth muscle cells (VSMCs). The purpose of this study was to investigate the effects of Mfn2 on the traffick- ing of intracellular cholesterol in the foam ceils derived from rat VSMCs (rVSMCs) and also to investigate the effects of Mfn2 on the expression of adenosine triphosphate-binding cassette subfamily A member 1 (ABCA1), adenosine triphosphate-binding cassette subfamily G member 1 (ABCG1) and peroxisome proliferator-activated receptor gamma (PPARy). The rVSMCs were co-cultured with oxi- dized low density lipoprotein (LDL, 80 ~tg/mL) to produce foam cells and cholesterol accumulation in cells. Before oxidized LDL treatment, different titers (20, 40 and 60 pfu/cell) of recombinant adenovirus containing Mfn2 gene (Adv-Mfn2) were added into the culture medium for 24 h to transfect the Mfn2 gene into the rVSMCs. Then the cells were harvested for analyses. The protein expression of Mfn2 was significantly higher in Adv-Mfn2-transfected group than in untransfected group (P〈0.05), and the ex- pression levels significantly increased when the titer of Adv-Mfn2 increased (P〈0.05). At 24 or 48 h af- ter oxidized LDL treatment, rVSMCs became irregular and their nuclei became larger, and their plasma abounded with red lipid droplets. However, the number of red lipid droplets was significantly decreased in Adv-Mfn2-transfected group as compared with untransfected group. At 48 h after oxidized LDL treatment, the intracellular cholesterol in rVSMCs was significantly increased (P〈0.05), but it was sig- nificantly decreased in Adv-Mfn2-transfected group as compared with untransfected group (P〈0.05), and it also significantly decreased when the titer of Adv-Mfn2 increased (P〈0.05). The mRNA and pro- tein expression levels of ABCA1 and ABCG1 were significantly increased in Adv-Mfn2-transfected group as compared with untransfected group (P〈0.05). Though the mRNA and protein expression levels of PPARy was not significantly increased (P〉0.05), the phosporylation levels of PPARy were signifi- cantly decreased in Adv-Mfn2-transfected group as compared with untransfected group (P〈0.05). These results suggest that the transfection of Adv-Mfn2 can significantly reduce intracellular cholesterol in oxidized LDL-induced rVSMCs possibly by decreasing PPAR'/phosporylation and then increasing pro- tein expression levels of ABCAI and ABCG1, which may be helpful to suppress the formation of foam cells.展开更多
文摘目的探讨金雀根和黄芪配伍对糖尿病肾病(DKD)大鼠的影响。方法将SD大鼠随机分为正常组、模型组、恩格列净组、金雀根组、黄芪组、配伍组,每组10只。采用单侧切除肾脏联合腹腔注射链脲佐菌素(STZ)复制DKD大鼠模型,造模成功后灌胃相应剂量药物8周。第0、4、8周检测24 h尿微量白蛋白(24 h U-mALB);ELISA法检测Scr、BUN、CysC、MDA水平及SOD活性;荧光探针法检测肾组织ROS表达;HE、PAS、Masson、PASM-Masson染色观察肾组织病理结构改变;免疫组织化学法检测肾组织NOX4、Drp1、MFN2、P62表达;Western blot法检测肾组织PINK1、MFN2、Parkin、LC3-Ⅱ/Ⅰ、P62、p-Drp1蛋白表达。结果与模型组比较,各给药组大鼠24 h U-mALB、BUN、Scr、CysC水平均降低(P<0.01),肾组织病理性结构损伤改善;血清和组织MDA水平降低(P<0.01),SOD活性升高(P<0.01);肾组织PINK1、MFN2、Parkin、LC3-Ⅱ/Ⅰ蛋白表达升高(P<0.05,P<0.01),p-Drp1、P62蛋白表达降低(P<0.01),其中配伍组作用优于单用药组(P<0.05,P<0.01)。结论金雀根和黄芪配伍可能是通过调控PINK1/MFN2/Parkin通路改善线粒体动力学,激活线粒体自噬,抑制氧化应激,减轻肾脏病理损伤,改善DKD大鼠肾功能。
文摘目的观察重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对缺血/再灌注损伤大鼠心肌细胞Mitofusin2(Mfn2)蛋白表达的影响及其抗心肌细胞凋亡的作用。方法选取成年SD大鼠35只,随机分为正常组(Normal),假手术组(Sham),缺血再灌注组(I/R),缺血再灌注EPO治疗组(I/R+EPO)。各组分别于再灌注3h和24h后,剪取心脏缺血/再灌注损伤区域,用脱氧核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测心肌细胞凋亡,免疫组化法检测Mfn2蛋白的表达。结果再灌注3h和24h后,与正常组和假手术组相比,I/R组Mfn2蛋白的表达和心肌细胞凋亡均显著增加;与I/R组相比,I/R+EPO组Mfn2蛋白的表达和心肌细胞凋亡均显著降低。结论EPO可以下调缺血再灌注损伤后心肌细胞Mfn2蛋白的表达,抑制心肌细胞的凋亡。
基金supported by the National Natural Science Foundation of China(No.30971244)
文摘Mitofusin2 (Mfn2) plays a pivotal role in the proliferation and apoptosis of vascular smooth muscle cells (VSMCs). The purpose of this study was to investigate the effects of Mfn2 on the traffick- ing of intracellular cholesterol in the foam ceils derived from rat VSMCs (rVSMCs) and also to investigate the effects of Mfn2 on the expression of adenosine triphosphate-binding cassette subfamily A member 1 (ABCA1), adenosine triphosphate-binding cassette subfamily G member 1 (ABCG1) and peroxisome proliferator-activated receptor gamma (PPARy). The rVSMCs were co-cultured with oxi- dized low density lipoprotein (LDL, 80 ~tg/mL) to produce foam cells and cholesterol accumulation in cells. Before oxidized LDL treatment, different titers (20, 40 and 60 pfu/cell) of recombinant adenovirus containing Mfn2 gene (Adv-Mfn2) were added into the culture medium for 24 h to transfect the Mfn2 gene into the rVSMCs. Then the cells were harvested for analyses. The protein expression of Mfn2 was significantly higher in Adv-Mfn2-transfected group than in untransfected group (P〈0.05), and the ex- pression levels significantly increased when the titer of Adv-Mfn2 increased (P〈0.05). At 24 or 48 h af- ter oxidized LDL treatment, rVSMCs became irregular and their nuclei became larger, and their plasma abounded with red lipid droplets. However, the number of red lipid droplets was significantly decreased in Adv-Mfn2-transfected group as compared with untransfected group. At 48 h after oxidized LDL treatment, the intracellular cholesterol in rVSMCs was significantly increased (P〈0.05), but it was sig- nificantly decreased in Adv-Mfn2-transfected group as compared with untransfected group (P〈0.05), and it also significantly decreased when the titer of Adv-Mfn2 increased (P〈0.05). The mRNA and pro- tein expression levels of ABCA1 and ABCG1 were significantly increased in Adv-Mfn2-transfected group as compared with untransfected group (P〈0.05). Though the mRNA and protein expression levels of PPARy was not significantly increased (P〉0.05), the phosporylation levels of PPARy were signifi- cantly decreased in Adv-Mfn2-transfected group as compared with untransfected group (P〈0.05). These results suggest that the transfection of Adv-Mfn2 can significantly reduce intracellular cholesterol in oxidized LDL-induced rVSMCs possibly by decreasing PPAR'/phosporylation and then increasing pro- tein expression levels of ABCAI and ABCG1, which may be helpful to suppress the formation of foam cells.