糖尿病肾病临床病理分期正常肾小球52kd72kdabcdabcd92kdabcd57kdabcdMMP1MMP9MMP3MMP2高糖对NHMC中MMP1,2,3,9表达的影响结果高糖环境下肾小球系膜细胞的增殖明显增加，而肝素能明显抑制肾小球系膜细胞的增殖高糖环境下肾小球系膜细胞MMPs的表达异常提示MMPs参与了细胞外基质的合成与降解的过程肝素能影响肾小球系膜细胞MMPs的表达，从而提示肝素对糖尿病肾病的肾保护作用是通过非抗凝作用而实现结论ACEI与ARB在肾脏病中的应用中南大学湘雅医院肾内科周巧玲背景糖尿病的主要并发症冠心病周围神经病变视网膜病变糖尿病肾病背景糖尿病肾病是糖尿病最严重的并发症之一最近研究显示，糖尿病肾病的发病率呈明显增加糖尿病肾病的发病机制目前并未完全明确2型糖尿病肾病的自然病程心血管死亡功能改变结构改变+血压升高微量白蛋白尿临床蛋白尿血肌酐升高ESRD5102030年临床2型糖尿病出现糖尿病肾脏血流动力学改变,肾小球滤过+肾小球基底膜增厚↑,系膜扩张↑,微血管改变+/- ESRD的发病率及其病因糖尿病发病率（国内）1980年（14省市30万人口）0.67％1994年（19省市21.35万人口）2.51％1996年（11省市42.75万人口）3.21％预计2025年全球3亿多元醇代谢通路的激活糖基化终末产物的产生葡萄糖转运蛋白的作用甘油二酯（DAG）－蛋白激酶C（PKC）信号传导通路激活糖代谢异常所致的DN的发生发展的因素目前关于糖尿病肾病发病机制的研究三高：高滤过、高灌注、高跨膜压代谢的改变生长因子(TGF-β,IL-6,PDGF)细胞外基质（ECM）合成增加遗传因素蛋白激酶C(PKC)丝裂原活化蛋白(MAP)糖基化终末产物(AGE)Diabetologia(2001)44:659~673HyperglycaemiaDAG(esp.PKCζ)ζAtypicalaPKCswidespreadProteinKinaseCIsoformsDiabetologia(2001)44:659~673HyperglycaemiaPKCζactivationECMaccumulationDIABETICNEPHROPATHY?TIMPand?MMP?基质合成基质降解稳态糖尿病肾病的机制研究合成降解细胞外基质糖尿病中促进ECM积聚的因素高糖TGF-β基质蛋白MMPs纤溶酶原活化物MMP活化纤溶酶的活性细胞外基质积聚TGF-β基质的降解？CellularandMolecularBiology45（1），123-135TIMPs参与细胞基质降解的四种酶天门冬氨酸酶半胱氨酸酶丝氨酸酶基质金属蛋白酶(MMPs)纤维化前状态基质合成基质降解正常间质基质蛋白正常小管基底膜蛋白异常基质蛋白蛋白多糖/糖蛋白多糖金属蛋白酶抑制剂纤溶酶原激活剂抑制剂糖基化作用,氧化作用.转谷氨酰胺酶交叉联系基质稳定Kidneyinternational,vol.58,suppl.77,pp.s81-87高糖对MMP活性影响简图表达纤溶酶活化活性基质降解高糖基质积聚肾小球系膜细胞MMP的主要类型MMP-2MT1-MMP膜型金属蛋白酶蛋白多糖，laminin，纤连蛋白，III,IV,V型胶原MMP-3,MMP-7,MMP-10基质降解素IV,V和VI型胶原、纤连蛋白、明胶MMP-2,MMP-9IV型胶原酶或明胶酶II和III型胶原MMP-1,MMP-8,MMP-13间质胶原酶酶作用底物名称MMP类型Kidneyinternational,vol.58,suppl.77,pp.s81-87Ⅰ期高滤过，肾小球体积增大GFR?MAP(-)BP(-)肾小球肥大，GBM和系膜正常Ⅱ期正常白蛋白尿?/(-)应激后??/(-)GBM增厚，MC增殖Ⅲ期微量白蛋白尿大致(-)??GBM增厚，ECM?Ⅳ期大量白蛋白尿??200ug/24h??GBM增厚，ECM??，小球荒废Ⅴ期ESRD??小球荒废面????硬化（广泛）糖尿病肾脏KW结节局灶性节段性肾小球硬化终末期肾病（硬化）高糖环境下正常人类肾小球系膜细胞中MMP表达及肝素对其表达的影响正常人类肾小球系膜细胞(NHMC)培养基(5mMDMEM,25mMDMEM)10%小牛血清(10%FCS)青霉素(100u/ml),链霉素(100mg/ml)肝素培养调节:37℃,5%CO2,95%空气细胞培养材料与方法细胞增殖测定WST-1法(波长450/600nm)Westernblot分析抗体1:MMP1,2,3,9抗体2:羊抗鼠IgG(辣根过氧化物酶)材料与方法高糖对NHMC增殖的影响结果肝素对NHMC增殖的影响（LG）结果结果肝素对NHMC增殖的影响（HG）Sheet3
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.00
.00
.00
.00
00.00
00.00
.00
.00
.00
.00
00.00
00.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
0.00
0.00
0.00
0.00
(ug/ml)
(OD540nm)
(ug/ml)
(OD540nm)
(ug/mi)
(OD540nm)
(ug/ml)
(OD540nm)
(%）
(OD540nm)
(%)
(OD540）
(Ethanol%)
(OD540nm)
(Ethanol%)
OD540nm)
(VitEug/ml)
(OD540)
(Heparinug/ml)
()D540nm)
(Heparinug/ml)
(OD540）
(VitEug/ml)
(OD540nm)
Hour
OD540nM
LG
HG
cellcultureunderLGandHG
LG
HG
NHMCCultureTime(hour)
OD（450/600nm）
HeparinConcentration(ug/ml)
OD(450/600nm)
HeparinConcentration(ug/ml)
OD(450/600nm)
肝素浓度(ug/ml)
OD(450/600nm)
.00
0.00
0.00
00.00
00.00
00.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
.00
Sheet3
Sheet2
Sheet1
图表1
DM
HTN
GN
CK
All
000.00
0.00
0.00
0.00
.00
00.00
000.00
0.00
0.00
0.00
.00
00.00
000.00
0.00
0.00
0.00
.00
00.00
000.00
0.00
0.00
0.00
.00
00.00
000.00
00.00
0.00
0.00
.00
00.00
000.00
00.00
0.00
0.00
.00
00.00
000.00
00.00
0.00
0.00
.00
00.00
000.00
00.00
0.00
0.00
.00
00.00
000.00
00.00
0.00
0.00
.00
00.00
000.00
00.00
0.00
0.00
.00
00.00
DM
HTN
GN
CK
All
Patients(n/millionpeoples)
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
000.00
0.00
0.00
0.00
.00
00.00
0.00
0.00
0.00
.00
00.00
0.00
0.00
0.00
.00
00.00
0.00
0.00
0.00
.00
00.00
00.00
0.00
0.00
.00
00.00
00.00
0.00
0.00
.00
00.00
00.00
0.00
0.00
.00
00.00
00.00
0.00
0.00
.00
00.00
00.00
0.00
0.00
.00
00.00
00.00
0.00
0.00
.00
00.00
|
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