目的:观察自发性高血压大鼠(spontaneously hypertensive rats,SHR)肾脏组织中Klotho、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)、基质金属蛋白酶组织抑制剂-1(tissue inhibitor of metalloproteinase-1,TIMP-1)和纤溶酶原激活物抑制剂-1(plasminogen activator inhibitor,PAI-1)基因的表达状况,探讨其在高血压肾间质纤维化中的作用及福辛普利和缬沙坦的肾保护作用机制. 方法:22周龄雄性SHR 20只随机分为4组(每组5只):高血压组(SHR组)、fosinopril组[Fos组,10 mg/( kg·d)灌胃],Valsartan组[Val组,50 mg/( kg·d)灌胃]和fosinopril+valsartan组[Fos+Val组,fosinopril 10 mg/( kg·d)+valsartan 50 mg/( kg·d)灌胃].22周龄雄性Wistar Kyoto大鼠(WKY组)5只作为对照组.用药物干预连续喂养8周,分别用RT-PCR和Western 印迹检测肾脏组织中Klotho,MMP-9,TIMP-1和PAI-1 mRNA和蛋白的表达水平. 结果:SHR组肾脏组织Klotho mRNA和蛋白表达较WKY组明显下调(P＜0.01),而MMP-9,TIMP-1和PAI-1mRNA和蛋白的表达较WKY组均明显上调(P＜0.01);用fosinopril和/或valsartan干预后,可上调Klotho mRNA在Fos,Fos+Val组(P＜0.01)和Val组(P＜0.05)的表达,上调Klotho 蛋白在Val,Fos+Val组(P＜0.01)和Fos组(P＜0.05)的表达.同时下调MMP-9,TIMP-1,PAI-1 mRNA和蛋白在Fos,Val,Fos+Val组的表达(P＜0.01).相关分析表明:Klotho mRNA和蛋白的表达与MMP-9,TIMP-1,PAI-1 mRNA (r值分别为-0.864,-0.725和-0.785,P＜0.01)和蛋白(r值分别为-0. 614,-0.579和-0.552,P＜0.05)的表达均呈负相关. 结论:高血压肾损害与抗衰老基因Klotho和基质降解相关基因MMP-9,TIMP-1,PAI-1关系密切;Klotho基因与MMP-9,TIMP-1和PAI-1基因的表达呈负相关.上调抗衰老基因Klotho表达的药物fosinopril 和/或valsartan可改善Klotho-MMPs/TIMPs-PAI-1的表达,这可能是其抗肾纤维化的重要机制.
目的:觀察自髮性高血壓大鼠(spontaneously hypertensive rats,SHR)腎髒組織中Klotho、基質金屬蛋白酶-9(matrix metalloproteinase-9,MMP-9)、基質金屬蛋白酶組織抑製劑-1(tissue inhibitor of metalloproteinase-1,TIMP-1)和纖溶酶原激活物抑製劑-1(plasminogen activator inhibitor,PAI-1)基因的錶達狀況,探討其在高血壓腎間質纖維化中的作用及福辛普利和纈沙坦的腎保護作用機製. 方法:22週齡雄性SHR 20隻隨機分為4組(每組5隻):高血壓組(SHR組)、fosinopril組[Fos組,10 mg/( kg·d)灌胃],Valsartan組[Val組,50 mg/( kg·d)灌胃]和fosinopril+valsartan組[Fos+Val組,fosinopril 10 mg/( kg·d)+valsartan 50 mg/( kg·d)灌胃].22週齡雄性Wistar Kyoto大鼠(WKY組)5隻作為對照組.用藥物榦預連續餵養8週,分彆用RT-PCR和Western 印跡檢測腎髒組織中Klotho,MMP-9,TIMP-1和PAI-1 mRNA和蛋白的錶達水平. 結果:SHR組腎髒組織Klotho mRNA和蛋白錶達較WKY組明顯下調(P＜0.01),而MMP-9,TIMP-1和PAI-1mRNA和蛋白的錶達較WKY組均明顯上調(P＜0.01);用fosinopril和/或valsartan榦預後,可上調Klotho mRNA在Fos,Fos+Val組(P＜0.01)和Val組(P＜0.05)的錶達,上調Klotho 蛋白在Val,Fos+Val組(P＜0.01)和Fos組(P＜0.05)的錶達.同時下調MMP-9,TIMP-1,PAI-1 mRNA和蛋白在Fos,Val,Fos+Val組的錶達(P＜0.01).相關分析錶明:Klotho mRNA和蛋白的錶達與MMP-9,TIMP-1,PAI-1 mRNA (r值分彆為-0.864,-0.725和-0.785,P＜0.01)和蛋白(r值分彆為-0. 614,-0.579和-0.552,P＜0.05)的錶達均呈負相關. 結論:高血壓腎損害與抗衰老基因Klotho和基質降解相關基因MMP-9,TIMP-1,PAI-1關繫密切;Klotho基因與MMP-9,TIMP-1和PAI-1基因的錶達呈負相關.上調抗衰老基因Klotho錶達的藥物fosinopril 和/或valsartan可改善Klotho-MMPs/TIMPs-PAI-1的錶達,這可能是其抗腎纖維化的重要機製.
목적:관찰자발성고혈압대서(spontaneously hypertensive rats,SHR)신장조직중Klotho、기질금속단백매-9(matrix metalloproteinase-9,MMP-9)、기질금속단백매조직억제제-1(tissue inhibitor of metalloproteinase-1,TIMP-1)화섬용매원격활물억제제-1(plasminogen activator inhibitor,PAI-1)기인적표체상황,탐토기재고혈압신간질섬유화중적작용급복신보리화힐사탄적신보호작용궤제. 방법:22주령웅성SHR 20지수궤분위4조(매조5지):고혈압조(SHR조)、fosinopril조[Fos조,10 mg/( kg·d)관위],Valsartan조[Val조,50 mg/( kg·d)관위]화fosinopril+valsartan조[Fos+Val조,fosinopril 10 mg/( kg·d)+valsartan 50 mg/( kg·d)관위].22주령웅성Wistar Kyoto대서(WKY조)5지작위대조조.용약물간예련속위양8주,분별용RT-PCR화Western 인적검측신장조직중Klotho,MMP-9,TIMP-1화PAI-1 mRNA화단백적표체수평. 결과:SHR조신장조직Klotho mRNA화단백표체교WKY조명현하조(P＜0.01),이MMP-9,TIMP-1화PAI-1mRNA화단백적표체교WKY조균명현상조(P＜0.01);용fosinopril화/혹valsartan간예후,가상조Klotho mRNA재Fos,Fos+Val조(P＜0.01)화Val조(P＜0.05)적표체,상조Klotho 단백재Val,Fos+Val조(P＜0.01)화Fos조(P＜0.05)적표체.동시하조MMP-9,TIMP-1,PAI-1 mRNA화단백재Fos,Val,Fos+Val조적표체(P＜0.01).상관분석표명:Klotho mRNA화단백적표체여MMP-9,TIMP-1,PAI-1 mRNA (r치분별위-0.864,-0.725화-0.785,P＜0.01)화단백(r치분별위-0. 614,-0.579화-0.552,P＜0.05)적표체균정부상관. 결론:고혈압신손해여항쇠로기인Klotho화기질강해상관기인MMP-9,TIMP-1,PAI-1관계밀절;Klotho기인여MMP-9,TIMP-1화PAI-1기인적표체정부상관.상조항쇠로기인Klotho표체적약물fosinopril 화/혹valsartan가개선Klotho-MMPs/TIMPs-PAI-1적표체,저가능시기항신섬유화적중요궤제.
Objective To determine the role of fosinopril and valsartan intervention in Klotho, matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1) and plasminogen activator inhibitor (PAI-1) gene expression in hypertensive renal interstitial fibrosis (RIF) in the kidney tissue of spontaneously hypertensive rats (SHR). Methods We randomly divided 20 male 22-week-old SHR into 4 groups (5 in each group):a hypertension group (SHR group), a fosinopril group ［Fos group, 10 mg/( kg·d) gavage］, a valsartan group ［Val group, 10 mg/( kg·d) gavage］， and a fosinopril plus valsartan group ［Fos + Val group, fosinopril 10 mg/( kg·d) + valsartan 50 mg/( kg·d) gavage］. Another five 22-week-old male Wistar Kyoto rats (WKY) were used as controls. Through monitoring the weight of the rats, tail artery pressure, 24-hour urine protein by fosinopril and/or valsartan intervention after the 8-week trial. RT-PCR and Western blot were used to detect the mRNA and protein expression of Klotho, MMP-9, TIMP-1, and PAI-1 in the kidneys.Results RT-PCR showed that in the SHR group, Klotho mRNA and protein expression were significantly decreased(P＜0.01), while mRNA and protein expression of MMP-9, TIMP-1, and PAI-1 were significantly higher compared with the WKY group(P＜0.01). With fosinopril and / or valsartan intervention, Klotho mRNA expression in the Fos group (P＜0.01), Fos + Val group (P＜0.01), Val group (P＜0.05), Klotho protein expression in the Fos group(P＜0.05), Fos + Val group (P＜0.05), Val group (P＜0.01), were significantly increased compared with those in the SHR group. The mRNA and protein expression of MMP-9, TIMP-1, and PAI-1 in the Fos group, Val group, and Fos + Val group were significantly lower than those in the SHR group (P＜0.01). The expression of Klotho mRNA had negative correlation with the expression of MMP-9 mRNA (r= -0.864, P<0.01), TIMP-1 mRNA (r=-0.725, P<0.01) and PAI-1 mRNA (r=-0.785, P<0.01). The Klotho protein expression had negative correlation with the expression of MMP-9 protein (r=-0.614, P<0.05), TIMP-1 protein (r=-0.579, P<0.05), and PAI-1 protein (r=-0.552, P<0.05). Conclusion Anti-aging gene Klotho and the genes related with extracellular matrix degradation gene MMP-9, TIMP-1, PAI-1 are involved in hypertensive renal injury. The expression of Klotho and MMP-9, TIMP-1, and PAI-1 is closely correlated. Fosinopril and valsartan which increase the Klotho mRNA and protein expression can alter the expression of Klotho-MMPs/TIMPs, which may be the main mechanism to prevent interstitial fibrosis.
