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12-脂氧化酶和血管紧张素Ⅱ对糖尿病大鼠肾小球p21、p27和p57表达的影响

Effects of 12-lipoxygenase and angiotensin Ⅱ on p21, p27 and p57 in rat diabetic glomeruli

摘要:

目的 探讨12-脂氧化酶(12-1ipoxygenase,12-LO)和血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)对细胞肥大相关的细胞周期素依赖激酶抑制蛋白p21、p27、p57表达的影响.方法 高糖处理肾小球系膜细胞24 h和48 h;雄性SD大鼠皮下包埋微型渗透泵,分别持续恒速注入12-LO代谢产物12(S)-HETE(1 mg· kg-1·d-1)1周和AngⅡ(400 ng·kg-1·min-1)2周.Western印迹检测p21、p27、p57蛋白表达.雄性SD大鼠分为普通饮食对照组、高脂饮食结合小剂量链脲菌素(STZ)诱导2型糖尿病(DN)组、DN+AngⅡ1型受体阻滞剂(ARB)或12-LO抑制剂(CDC)治疗组,分别给药6周和8周,采用ELISA法检测尿白蛋白,Western印迹检测p21、p27、p57蛋白表达;PAS染色观察肾小球病变程度,免疫组织化学法检测肾小球内p21、p27的表达.结果 与相应正常对照组比较,高糖组系膜细胞内p21、p27蛋白表达增加(均P<0.05),AngⅡ组大鼠肾小球内p27蛋白表达增加(P<0.05),12(S)-HETE组大鼠肾小球内p21、p27蛋白表达增加(均P< 0.05),其他周期素依赖激酶抑制蛋白表达差异均无统计学意义.与对照组比较,DN组大鼠血糖、肾重/体重及尿蛋白及肾小球内p21、p27蛋白表达均增加(均P<0.05),p57无明显变化(P>0.05);与DN组比较,ARB或CDC治疗组大鼠的血糖无明显变化,但肾重/体重比及尿白蛋白均降低(均P< 0.05),肾小球毛细血管袢肥大及系膜增生均减少,CDC组肾小球内p21和p27蛋白表达均降低(均P<0.05),但ARB组肾小球仅p27蛋白表达降低(P<0.05).结论 12-LO和AngⅡ对p21、p27和p57的影响不同,12-LO可诱导DN肾小球内p21和p27表达上调,但AngⅡ仅诱导DN肾小球内p27表达上调.

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abstracts:

Objective To investigate the effects of 12-lipoxygenase (12-LO) and angiotensin Ⅱ (Ang Ⅱ) on the CIP/KIP family of cyclin-dependent kinase inhibitors (CKIs) p21,p27 and p57 related to cell hypertrophy.Methods Mesangial cells were treated with high glucose for 24 hours and 48 hours respectively.12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE] and Ang Ⅱ were infused to rats by osmotic mini-pump for 1 week and 2 weeks respectively.Rats fed high fat diet were received low dose streptozotocin (STZ) to make type 2 diabetes (DN).The rats were divided into normal control group,DN group,DN+Ang Ⅱ type 1 receptor blocker (ARB) group or 12-LO inhibitor (CDC) group.DN+ARB rats were treated by losartan for 6 weeks,and DN+CDC rats were treated for 8 weeks.Urine albumin and protein expressions of p21,p27 and p57 were detected by ELISA and Western blotting respectively.Glomeruli injury and expressions of p21 and p27 were detected by PAS staining and immunohistochemistry respectively.Results High glucose increased p21 and p27 protein expression in mesangial cells significantly compared with the relative control (all P < 0.05),but had no effect on p57.Ang Ⅱ increased p27 protein expression in gloneruli significantly (P < 0.05),but had no effect on p21 and p57 protein expression.12(S)-HETE increased both p21 and p27 protein expression in glomeruli significantly (all P < 0.05),but had no effect on p57 protein expression.Blood glucose,kidney/body weight,urinary protein,and glomerular p21 and p27 protein expressions were increased in DN group (all P < 0.05) compared with those in control group,with little change of p57 protein expression (P < 0.05).Moreover,glomerular hypertrophy and extra cellular matrix accumulation were observed in DN group.However,urine protein,kidney/body weight,renal injury,but not blood glucose,were decreased in DN+ARB group and DN+CDC group compared with DN group respectively (P< 0.05).Further DN+CDC rats had decreased both p21 and p27 protein expressions in glomeruli,but DN+ ARB rats only had decreased p27 protein expression (all P < 0.05).Conclusions 12-LO may induce both p21 and p27 protein expression in DN glomeruli,but Ang Ⅱ may induce only p27 expression.

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作者: 臧崇森 [1] 王婉宁 [1] 贾冶 [1] 马福哲 [1] 孙韬 [1] 孙珉丹 [1] 孙伟霞 [1] 远航 [1] 许钟镐 [1]
期刊: 《中华肾脏病杂志》2017年33卷2期 132-139页 ISTICPKUCSCD
栏目名称: 基础研究
DOI: 10.3760/cma.j.issn.1001-7097.2017.02.009
发布时间: 2017-05-03
基金项目:
National Natural Science Foundation of China Jilin Science Project (20140204020YY)国家自然科学基金 吉林省科技厅科研项目
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