中华肾脏病杂志
中華腎髒病雜誌
중화신장병잡지
2009年
5期
381-386
,共6页
许钟镐%贾冶%崔英春%吴曼%马福哲%许圣淳%郭桥艳%苗里宁
許鐘鎬%賈冶%崔英春%吳曼%馬福哲%許聖淳%郭橋豔%苗裏寧
허종호%가야%최영춘%오만%마복철%허골순%곽교염%묘리저
脂氧合酶%糖尿病肾病%p38丝裂原活化蛋白激酶类%肾小球系膜细胞%p27Kip1%肾小球
脂氧閤酶%糖尿病腎病%p38絲裂原活化蛋白激酶類%腎小毬繫膜細胞%p27Kip1%腎小毬
지양합매%당뇨병신병%p38사렬원활화단백격매류%신소구계막세포%p27Kip1%신소구
Lipoxygenase%Diabetic nephropathies%p38 mitogen-activated proteinkinases%Mesangial cells%p27Kip1%Kidney glomerulus
目的 探讨12-脂氧化酶(12-LO)对糖尿病肾病肾小球内p27Kip1表达的影响.方法 在有或无p38 MAPK(p38)抑制剂(SB203580,1 μmol/L)的条件下,用12-LO作用产物12羟二十烷四烯酸[12(S)-HETE,10-7mmol/L]刺激系膜细胞24 h.雄性SD大鼠分为普通饮食对照组、普通饮食+12-LO抑制剂(CDC,8 mg/kg,3次/周,皮下注射)处理组、高脂饮食结合小剂量链脲菌素(STZ,35 mg/kg,腹腔注射)诱导2型糖尿病组、高脂饮食结合小剂量STZ诱导2型糖尿病+CDC处理组,连续皮下注射CDC 1个月.野生型和12-LO基因敲除C57BL/6小鼠随机分成野生型对照组、12-LO基因敲除组、野生型STZ(200 mg/kg,腹腔注射)诱导1型糖尿病组、12-LO基因敲除STZ诱导1型糖尿病组,饲养16周.实验结束后收集尿、血液、提取肾脏,用系列过筛方法 分离肾小球.Western印迹和免疫组织化学方法 检测p38活性和p27Kip1蛋白表达的变化.结果 抑制p38活性可有效阻断12(S)-HETE诱导的系膜细胞内p27Kip1的表达(P<0.01).CDC可抑制2型糖尿病大鼠肾小球体积增大,降低尿白蛋白量(24 h),阻止肾小球内p38活性和p27Kip1蛋白表达增加,与CDC未处理2型糖尿病大鼠比较差异均有统计学意义(均P<0.01).与野生型糖尿病小鼠比较,12-LO基因敲除糖尿病小鼠p38活性、p27Kip1蛋白表达及细胞外基质积聚显著减少,差异有统计学意义(均P<0.01).结论 12-LO可通过p38通路上调糖尿病肾小球内p27Kip1的表达.
目的 探討12-脂氧化酶(12-LO)對糖尿病腎病腎小毬內p27Kip1錶達的影響.方法 在有或無p38 MAPK(p38)抑製劑(SB203580,1 μmol/L)的條件下,用12-LO作用產物12羥二十烷四烯痠[12(S)-HETE,10-7mmol/L]刺激繫膜細胞24 h.雄性SD大鼠分為普通飲食對照組、普通飲食+12-LO抑製劑(CDC,8 mg/kg,3次/週,皮下註射)處理組、高脂飲食結閤小劑量鏈脲菌素(STZ,35 mg/kg,腹腔註射)誘導2型糖尿病組、高脂飲食結閤小劑量STZ誘導2型糖尿病+CDC處理組,連續皮下註射CDC 1箇月.野生型和12-LO基因敲除C57BL/6小鼠隨機分成野生型對照組、12-LO基因敲除組、野生型STZ(200 mg/kg,腹腔註射)誘導1型糖尿病組、12-LO基因敲除STZ誘導1型糖尿病組,飼養16週.實驗結束後收集尿、血液、提取腎髒,用繫列過篩方法 分離腎小毬.Western印跡和免疫組織化學方法 檢測p38活性和p27Kip1蛋白錶達的變化.結果 抑製p38活性可有效阻斷12(S)-HETE誘導的繫膜細胞內p27Kip1的錶達(P<0.01).CDC可抑製2型糖尿病大鼠腎小毬體積增大,降低尿白蛋白量(24 h),阻止腎小毬內p38活性和p27Kip1蛋白錶達增加,與CDC未處理2型糖尿病大鼠比較差異均有統計學意義(均P<0.01).與野生型糖尿病小鼠比較,12-LO基因敲除糖尿病小鼠p38活性、p27Kip1蛋白錶達及細胞外基質積聚顯著減少,差異有統計學意義(均P<0.01).結論 12-LO可通過p38通路上調糖尿病腎小毬內p27Kip1的錶達.
목적 탐토12-지양화매(12-LO)대당뇨병신병신소구내p27Kip1표체적영향.방법 재유혹무p38 MAPK(p38)억제제(SB203580,1 μmol/L)적조건하,용12-LO작용산물12간이십완사희산[12(S)-HETE,10-7mmol/L]자격계막세포24 h.웅성SD대서분위보통음식대조조、보통음식+12-LO억제제(CDC,8 mg/kg,3차/주,피하주사)처리조、고지음식결합소제량련뇨균소(STZ,35 mg/kg,복강주사)유도2형당뇨병조、고지음식결합소제량STZ유도2형당뇨병+CDC처리조,련속피하주사CDC 1개월.야생형화12-LO기인고제C57BL/6소서수궤분성야생형대조조、12-LO기인고제조、야생형STZ(200 mg/kg,복강주사)유도1형당뇨병조、12-LO기인고제STZ유도1형당뇨병조,사양16주.실험결속후수집뇨、혈액、제취신장,용계렬과사방법 분리신소구.Western인적화면역조직화학방법 검측p38활성화p27Kip1단백표체적변화.결과 억제p38활성가유효조단12(S)-HETE유도적계막세포내p27Kip1적표체(P<0.01).CDC가억제2형당뇨병대서신소구체적증대,강저뇨백단백량(24 h),조지신소구내p38활성화p27Kip1단백표체증가,여CDC미처리2형당뇨병대서비교차이균유통계학의의(균P<0.01).여야생형당뇨병소서비교,12-LO기인고제당뇨병소서p38활성、p27Kip1단백표체급세포외기질적취현저감소,차이유통계학의의(균P<0.01).결론 12-LO가통과p38통로상조당뇨병신소구내p27Kip1적표체.
Objective To investigate the effect of 12-lipoxygenase(12-LO) on the p27Kip1 expression in diabetic glomeruli. Methods Mesangial cells were exposed to 12-LO product 12 (S)-HETE (10-7 mmol/L) with or without p38 MAPK (p38) inhibitor (SB203580, 1 μmol/L) for 24 hours. Rats fed with high fat diet received low dose streptozotoein (ST-Z, 35 mg/kg, IP injection) to develop type 2 diabetes and were divided into 2 groups: low dose STZ, low dose STZ+12-LO inhibitor cinnamyl-3,4-dihydroxy-α-cynanocinnamate (CDC, 8 mg/kg) treatment. Rats fed with regular chow were divided into two groups: controls, CDC treatment. The rats received injection of CDC or vehicle subcutaneously in the hind leg. CDC or vehicle injection was performed three times weekly on alternate days. All the rats were sacrificed after 4 weeks, Wild type and 12-LO knockout C57BL/6 mice were divided into 4 groups: wild type control, 12-LO knockout, STZ-induced wild type type 1 diabetes and STZ-induced 12-LO knockout type 1 diabetes. All the mice were sacrificed after 16 weeks. Urine, blood, kidney cortical tissue and isolated glomeruli by sieving method were collected at the end of study respectively. Western blot and immunohistochemistry for target protein were performed respectively. Results Inhibition of p38 activation could significantly reduce p27Kip1 expression induced by 12 (S)-HETE in mesangial cells (P<0.01). Increased glomerular volume, microalbuminuria, elevated glomeluli p38 activation, p27Kip1 expresssion in type 2 diabetic glomeruli was decreased after CDC treatment (P<0.01). Compared with wild type diabetic mice, glomerular p38 activation, p27Kip1 exprcsssion and extracellular matrix accumulation in the 12-LO knockout diabetic mice were significantly decreased (P <0.01, respectively). Conclusions 12-LO induces p27kipl expression via p38 pathway in diabetic glomeruli.
