中华内分泌代谢杂志
中華內分泌代謝雜誌
중화내분비대사잡지
CHINESE JOURNAL OF ENDOCRINOLOGY AND METABOLISM
2013年
10期
849-852
,共4页
糖尿病肾病%1,25-二羟维生素D3%HK-2细胞%解耦联蛋白2%氧化应激
糖尿病腎病%1,25-二羥維生素D3%HK-2細胞%解耦聯蛋白2%氧化應激
당뇨병신병%1,25-이간유생소D3%HK-2세포%해우련단백2%양화응격
Diabetic nephropathy%1,25-dihydroxyvitamin D3%Human renal tubular epithelial cells%Uncoupling protein 2%Oxidative stress
体外培养的HK-2细胞分为正常对照组(NG组,5.5 mmol/L D-葡萄糖)、高糖处理组(HG组,30 mmol/L D-葡萄糖)、高渗对照组(MG组,5.5 mmol/L D-葡萄糖+24.5 mmol/L D-甘露醇)、高糖培养液中又加入不同浓度的1,25-二羟维生素D3[1,25-(OH)2D3]组(V1～V3组)、N-乙酰半胱氨酸药效对照组(NAC组,30 mmoL/L D-葡萄糖+1.0 mmol/L N-乙酰半胱氨酸)和无水乙醇溶剂对照组(SG组,30 mmol/LD-葡萄糖+6.86×10-2 mol/L乙醇).检测细胞活性氧簇荧光强度、线粒体膜电位、解耦联蛋白2(UCP2)mRNA、蛋白表达、总超氧化物歧化酶(SOD)活力和丙二醛水平.结果HG组细胞的线粒体膜电位明显低于NG组(P＜0.01),1,25-(OH)2D3处理后细胞的线粒体膜电位与HG组比较显著下降(均P＜0.01)；HG组总SOD活力显著低于NG组(P＜0.01),丙二醛含量明显高于NG组(P＜0.01),而1,25-(OH)2D3处理后细胞总SOD活力较HG组显著增高(P＜0.05),丙二醛水平显著低于HG组(P＜0.01)；HG组UCP2 mRNA与蛋白表达显著高于NG组(P＜0.05),而1,25-(OH)2D3处理后细胞UCP2的表达与HG组比较显著降低(P＜0.01).结果表明高糖可诱导体外培养的HK-2细胞氧化应激损伤；1,25-(OH)2D3可能通过降低线粒体膜电位、活性氧簇生成及调节胞内UCP2表达抑制高糖诱导的氧化应激反应.
體外培養的HK-2細胞分為正常對照組(NG組,5.5 mmol/L D-葡萄糖)、高糖處理組(HG組,30 mmol/L D-葡萄糖)、高滲對照組(MG組,5.5 mmol/L D-葡萄糖+24.5 mmol/L D-甘露醇)、高糖培養液中又加入不同濃度的1,25-二羥維生素D3[1,25-(OH)2D3]組(V1～V3組)、N-乙酰半胱氨痠藥效對照組(NAC組,30 mmoL/L D-葡萄糖+1.0 mmol/L N-乙酰半胱氨痠)和無水乙醇溶劑對照組(SG組,30 mmol/LD-葡萄糖+6.86×10-2 mol/L乙醇).檢測細胞活性氧簇熒光彊度、線粒體膜電位、解耦聯蛋白2(UCP2)mRNA、蛋白錶達、總超氧化物歧化酶(SOD)活力和丙二醛水平.結果HG組細胞的線粒體膜電位明顯低于NG組(P＜0.01),1,25-(OH)2D3處理後細胞的線粒體膜電位與HG組比較顯著下降(均P＜0.01)；HG組總SOD活力顯著低于NG組(P＜0.01),丙二醛含量明顯高于NG組(P＜0.01),而1,25-(OH)2D3處理後細胞總SOD活力較HG組顯著增高(P＜0.05),丙二醛水平顯著低于HG組(P＜0.01)；HG組UCP2 mRNA與蛋白錶達顯著高于NG組(P＜0.05),而1,25-(OH)2D3處理後細胞UCP2的錶達與HG組比較顯著降低(P＜0.01).結果錶明高糖可誘導體外培養的HK-2細胞氧化應激損傷；1,25-(OH)2D3可能通過降低線粒體膜電位、活性氧簇生成及調節胞內UCP2錶達抑製高糖誘導的氧化應激反應.
체외배양적HK-2세포분위정상대조조(NG조,5.5 mmol/L D-포도당)、고당처리조(HG조,30 mmol/L D-포도당)、고삼대조조(MG조,5.5 mmol/L D-포도당+24.5 mmol/L D-감로순)、고당배양액중우가입불동농도적1,25-이간유생소D3[1,25-(OH)2D3]조(V1～V3조)、N-을선반광안산약효대조조(NAC조,30 mmoL/L D-포도당+1.0 mmol/L N-을선반광안산)화무수을순용제대조조(SG조,30 mmol/LD-포도당+6.86×10-2 mol/L을순).검측세포활성양족형광강도、선립체막전위、해우련단백2(UCP2)mRNA、단백표체、총초양화물기화매(SOD)활력화병이철수평.결과HG조세포적선립체막전위명현저우NG조(P＜0.01),1,25-(OH)2D3처리후세포적선립체막전위여HG조비교현저하강(균P＜0.01)；HG조총SOD활력현저저우NG조(P＜0.01),병이철함량명현고우NG조(P＜0.01),이1,25-(OH)2D3처리후세포총SOD활력교HG조현저증고(P＜0.05),병이철수평현저저우HG조(P＜0.01)；HG조UCP2 mRNA여단백표체현저고우NG조(P＜0.05),이1,25-(OH)2D3처리후세포UCP2적표체여HG조비교현저강저(P＜0.01).결과표명고당가유도체외배양적HK-2세포양화응격손상；1,25-(OH)2D3가능통과강저선립체막전위、활성양족생성급조절포내UCP2표체억제고당유도적양화응격반응.
[Summary] The HK-2 cells with different culture media were divided into normal glucose group (NG group,5.5 mmol/L D-glucose) ; high glucose group (HG group,30 mmol/L D-glucose) ; mannitol group (MG group,5.5mmol/L D-glucose+24.5 mmol/L mannitol) ; 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] groups (V1-V3 group)which were exposed to medium containing 30 mmol/L D-glucose and different concentrations of 1,25-(OH)2D3 ;Nethyl-cysteim control group (NAC group,30 mmoL/L D-glucose + 1.0 mmol/L N-Nethyl-cysteim) ; and ethanol control group(SG group,30 mmol/L D-glucose+6.86 × 10-2 mol/L ethanol).The level of intracellular reactive oxygen species,mitochondrial membrane potential,activity of total-superoxide dismutase (T-SOD),level of malondialdehyde,expression of UCP2 mRNA and protein in HK-2 cells were detected.Compared with NG group,the mitochondrial membrane potential significantly decreased in HG group (P＜0.01),and the mitochondrial membrane potential in V group was lower than that in HG group(P＜0.01).The activity ofT-SOD in HG group was significantly lower than that in NG group(P＜0.01),while its level of malondialdehyde was significantly higher than that in NG group(P＜0.01).Compared with HG group,the activity of T-SOD in V groups was significantly increased (P＜0.05)and the level of malondialdehyde in these groups significantly decreased (P＜0.01).The mRNA expression of UCP2 in HG group was increased significantly in comparison with NG group (P ＜ 0.05) and the expression in V groups was significantly decreased in comparison with HG group (P＜0.01).The results suggest that 1,25-(OH)2D3 could reduce the mitochondrial membrane potential,the production of reactive oxygen species,and regulate the expression of UCP2 in order to suppress the oxidative stress induced by high glucose.
