Our previous study showed the early molecular responses of bone in response to obstructive nephropathy in a unilateral ureteral obstruction （UUO） mouse model. Here, we addressed the changes in trabecular bone proper...Our previous study showed the early molecular responses of bone in response to obstructive nephropathy in a unilateral ureteral obstruction （UUO） mouse model. Here, we addressed the changes in trabecular bone properties at greater trochanter, the proximal and the distal metaphysis of femur in UUO mice. The male mice were subjected to UUO （n= 10） or sham operation （n= 10）. All mice were killed on day 7 after the surgical operation. The micro-computed tomography （micro-CT） analysis for different femoral trabecular bone sites demonstrated pathological alterations of trabecular bone mass and micro-networks at greater trochanter as shown by decreases in bone mineral density/bone volume （P〈O.05） and trabecular number （P〈O.05） and increases in trabecular separation （P〈O.01） and bone surface/bone volume （P〈O.05） in UUO mice. The present study demonstrates that UUO-induced unilateral obstructivenephropathy has markedly detrimental effects on the trabecular trochanter of the femur.展开更多
目的探讨硫化氢(hydrogen sulfide,H 2S)改善单侧输尿管梗阻(unilateral ureteral obstruction,UUO)小鼠肾脏纤维化的作用机制。方法采用C57BL/6小鼠单侧输尿管结扎建立肾脏纤维化小鼠模型。随机分为假手术(Sham)组、UUO组和UUO+硫氢化...目的探讨硫化氢(hydrogen sulfide,H 2S)改善单侧输尿管梗阻(unilateral ureteral obstruction,UUO)小鼠肾脏纤维化的作用机制。方法采用C57BL/6小鼠单侧输尿管结扎建立肾脏纤维化小鼠模型。随机分为假手术(Sham)组、UUO组和UUO+硫氢化钠(NaHS)组。ELISA法检测血清H 2S浓度;HE染色和Masson染色观察肾脏病理改变;免疫组织化学和Western blot检测Klotho和DNA甲基转移酶1(DNA methyltransferase 1,DNMT1)表达;RT-PCR检测双加氧酶(ten-eleven translocation,TET)表达;比色法检测TET活性;焦磷酸测序检测肾组织Klotho甲基化水平;羟甲基化DNA免疫共沉淀联合实时定量PCR法检测肾组织Klotho羟甲基化水平。结果与Sham组相比,UUO组血清H 2S浓度显著降低[(5.18±0.34)μmol/L vs.(4.23±0.21)μmol/L, P <0.05]。NaHS显著减轻UUO模型小鼠的肾小管间质纤维化( P <0.01),下调α平滑肌肌动蛋白( P <0.05)和纤连蛋白( P <0.05)表达,同时上调UUO小鼠肾组织Klotho表达( P <0.05),下调DNMT1表达( P <0.01),升高TET活性[(0.03±0.01) ng·min^-1·mg^-1 vs.(0.43±0.08) ng·min^-1·mg^-1 , P <0.05],降低 Klotho 基因启动子甲基化水平(11.83%±0.53% vs .7.39%±0.70%, P <0.01),升高 Klotho 基因启动子羟甲基化水平( P <0.05)。结论H 2S可通过增强TET活性,诱导 Klotho 基因启动子去甲基化,上调 Klotho 基因表达,从而减轻UUO小鼠肾脏纤维化。展开更多
文摘Our previous study showed the early molecular responses of bone in response to obstructive nephropathy in a unilateral ureteral obstruction （UUO） mouse model. Here, we addressed the changes in trabecular bone properties at greater trochanter, the proximal and the distal metaphysis of femur in UUO mice. The male mice were subjected to UUO （n= 10） or sham operation （n= 10）. All mice were killed on day 7 after the surgical operation. The micro-computed tomography （micro-CT） analysis for different femoral trabecular bone sites demonstrated pathological alterations of trabecular bone mass and micro-networks at greater trochanter as shown by decreases in bone mineral density/bone volume （P〈O.05） and trabecular number （P〈O.05） and increases in trabecular separation （P〈O.01） and bone surface/bone volume （P〈O.05） in UUO mice. The present study demonstrates that UUO-induced unilateral obstructivenephropathy has markedly detrimental effects on the trabecular trochanter of the femur.
文摘目的探讨硫化氢(hydrogen sulfide,H 2S)改善单侧输尿管梗阻(unilateral ureteral obstruction,UUO)小鼠肾脏纤维化的作用机制。方法采用C57BL/6小鼠单侧输尿管结扎建立肾脏纤维化小鼠模型。随机分为假手术(Sham)组、UUO组和UUO+硫氢化钠(NaHS)组。ELISA法检测血清H 2S浓度;HE染色和Masson染色观察肾脏病理改变;免疫组织化学和Western blot检测Klotho和DNA甲基转移酶1(DNA methyltransferase 1,DNMT1)表达;RT-PCR检测双加氧酶(ten-eleven translocation,TET)表达;比色法检测TET活性;焦磷酸测序检测肾组织Klotho甲基化水平;羟甲基化DNA免疫共沉淀联合实时定量PCR法检测肾组织Klotho羟甲基化水平。结果与Sham组相比,UUO组血清H 2S浓度显著降低[(5.18±0.34)μmol/L vs.(4.23±0.21)μmol/L, P <0.05]。NaHS显著减轻UUO模型小鼠的肾小管间质纤维化( P <0.01),下调α平滑肌肌动蛋白( P <0.05)和纤连蛋白( P <0.05)表达,同时上调UUO小鼠肾组织Klotho表达( P <0.05),下调DNMT1表达( P <0.01),升高TET活性[(0.03±0.01) ng·min^-1·mg^-1 vs.(0.43±0.08) ng·min^-1·mg^-1 , P <0.05],降低 Klotho 基因启动子甲基化水平(11.83%±0.53% vs .7.39%±0.70%, P <0.01),升高 Klotho 基因启动子羟甲基化水平( P <0.05)。结论H 2S可通过增强TET活性,诱导 Klotho 基因启动子去甲基化,上调 Klotho 基因表达,从而减轻UUO小鼠肾脏纤维化。
