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
目的探讨2型糖尿病肾病（diabebes nephropathy，DN）血清氧化型低密度脂蛋白（oxidized low density lipopro-tein，ox—LDL）与尿清蛋白之间的关系。方法根据尿清蛋白排泄率（urinary albumin excretion rate，UAER）将103例2型糖尿...目的探讨2型糖尿病肾病（diabebes nephropathy，DN）血清氧化型低密度脂蛋白（oxidized low density lipopro-tein，ox—LDL）与尿清蛋白之间的关系。方法根据尿清蛋白排泄率（urinary albumin excretion rate，UAER）将103例2型糖尿病患者分为单纯糖尿病（A）组、早期糖尿病肾病（B）组和临床糖尿病肾病（C）组，另选30例健康体检者作为对照（D）组，采用酶联免疫吸附试验双抗体夹心法检测ox—LDL。结果血清ox—LDL水平在A，B，C纽与对照组之间比较差异有统计学意义（P〈0．05），B组和C组显著高于A组（P〈0．05），C组显著高于B组（P〈0．05）。ox-LDL与UAER高度相关（r=0．81，P〈0．01）。结论DN患者血清ox—LDL水平随着尿微量清蛋白的增加而升高，当尿微量清蛋白还在正常水平时，ox—LDL的水平已较对照组明显升高，提示对于DN的早期诊断，ox—LDL可能较尿微量清蛋白更敏感；ox-LDL可作为DN敏感的早期诊断指标。展开更多
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.
文摘目的探讨2型糖尿病肾病（diabebes nephropathy，DN）血清氧化型低密度脂蛋白（oxidized low density lipopro-tein，ox—LDL）与尿清蛋白之间的关系。方法根据尿清蛋白排泄率（urinary albumin excretion rate，UAER）将103例2型糖尿病患者分为单纯糖尿病（A）组、早期糖尿病肾病（B）组和临床糖尿病肾病（C）组，另选30例健康体检者作为对照（D）组，采用酶联免疫吸附试验双抗体夹心法检测ox—LDL。结果血清ox—LDL水平在A，B，C纽与对照组之间比较差异有统计学意义（P〈0．05），B组和C组显著高于A组（P〈0．05），C组显著高于B组（P〈0．05）。ox-LDL与UAER高度相关（r=0．81，P〈0．01）。结论DN患者血清ox—LDL水平随着尿微量清蛋白的增加而升高，当尿微量清蛋白还在正常水平时，ox—LDL的水平已较对照组明显升高，提示对于DN的早期诊断，ox—LDL可能较尿微量清蛋白更敏感；ox-LDL可作为DN敏感的早期诊断指标。
