法舒地尔对高糖培养肾小管上皮细胞增殖和纤维化的影响.pdf

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1、第44卷第1O期温州医科大学学报v01．44NO．102014年1O月JournalofWenzhouMedicalUniversityoct．2O14籀法舒地尔对高糖培养肾小管上皮细胞增殖和纤维化的影响顾玲佳，倪连松(1．台州市第一人民医院内分泌科，浙江台卅f318020；2．温卅f医科大学附属第一医院内分泌科浙江温州325015)[摘要]目的：探讨法舒地尔(Fasudil)对高糖培养的肾小管上皮细胞(HK．2)增殖和纤维化的影响。方法：将HK．2细胞分为正常对照组(NG组，葡萄糖浓度5．5mmol／L)、高张组(HM组，葡萄糖浓度5．5mmol／L+甘露

2、醇54．5mmol／L)、高糖组(HG组，葡萄糖浓度60mmol／L)、高糖+小剂量法舒地尔干预组[FA(5)组，D．葡萄糖60mmol／L+法舒地尔5~mol／L]、高糖+中剂量法舒地尔干预组[FA(10)组，D．葡萄糖60mmol／L+法舒地尔10gmol／L]；⑥高糖+大剂量法舒地尔干预组[FA(20)组，D一葡萄糖60mmol／L+法舒地尔2o~moZ／L】。四甲基偶氮唑盐(MTT)法测定HK一2细胞增殖；免疫共沉淀法检测P—MYPT1．Thr853；Western印迹法检测结缔组织生长因子(CTGF)的表达；ELISA法检测细胞培养上清液转化生长因

3、子p(TGF．p)和纤维连接蛋白(Fn)的含量。结果：与NG组比，HG组HK一2细胞出现增殖增加，p-MYPTI．Thr853、TGF．p、CTGF和Fn的表达增加。与HG组比，FA(5)组、FA(10)组、FA(20)组均抑制了HK一2细胞的过度增殖，P—MYPT1．Thr853、TGF．pCTGF和Fn的表达均减少。结论：法舒地尔可能通过抑制Rho激酶活性抑制高糖培养的肾小管上皮细胞增殖和纤维化。[关键词]法舒地尔；肾小管上皮细胞；纤维化；糖尿病肾病[中图分类号]R965．2[文献标志码]A[文章编号]1000-2138(2014)10-0723一o5I

4、nfluenceoffasudilontheproliferationandfibrosisofrenaltubularepithelialcellsincubatedwithhighglucoseGULing／ia1．NILiansongz．i．DepartmentofEndocrinology,TaizhouFirstPeopleHospital,Ta~hou。318020；2．DepartmentofEndocrinology,theFirstAfiliatedHospitalofWenzhouMedicalUniversity,Wenzhou。325

5、015objective：Toinvestigatetheinfluenceoffasudilontheproliferationandfibrosisofhumanrenaltubularepithelialcellslncubatedwithhighglucose．Methods：HK一2cellsweredividedIntonormalcontrolgroup(D-glucose5．5mmol／L)，hightensiongroup(D—glucose5．5mmol／L+54．5mmol／Lmannito1)，highglucosegroup(D—g

6、lucose60mmol／L)，highglucose+fasudiltreatmentgroup(theconcentrationsoffasudilwere5，10and20gmol／L)．CellproliferationwasassessedwithMTTassay~Changesinthep-MYPT1一Thr853weredetectedwithCO—immunoprecipitationassay~Theexpressionofconnectivetissuegrowthfactor(CTGF)wasdetectedwithwesternblo

7、t，theproteincontentoftransforminggrowthfactor-131(TGF一131)andfibronectin(Fn)inthesuperna—tantweredetectedbyELISA．Results：Comparedwithnormalcontrolgroup，cellproliferationandtheexpres—sionofp-MYPT1-Thr853，TGF—pl，CTGFandFnweresignificantlyincreasedinhighglucosegroup．Comparedwithhighgl

8、ucosegroup，cellproliferati