The aim of this study was to investigate the mechanism by which a diet inducing high hyperhomocysteinemia(HHcy)leads to the deterioration of erectile function in rats and whether this is inhibited by expression of the...The aim of this study was to investigate the mechanism by which a diet inducing high hyperhomocysteinemia(HHcy)leads to the deterioration of erectile function in rats and whether this is inhibited by expression of the human tissue kallikrein-1(hKLKl)gene.We established a rat model of HHcy by feeding methionine(Met)-rich diets to male Sprague-Dawley(SD)rats.Male wild-type SD rats(WTRs)and transgenic rats harboring the hKLKl gene(TGRs)were fed a normal diet until 10 weeks of age.Then,30 WTRs were randomly divided into three groups as follows:the control(n=10)group,the low-dose(4%Met,n=10)group,and the high-dose(7%Met,n=10)group.Another 10 age-matched TGRs were fed the high-dose diet and designated as the TGR+7%Met group.After 30 days,in all four groups,erectile function was measured and penile tissues were harvested to determine oxidative stress,endothelial cell content,and penis fibrosis.Compared with the 7%Met group,the TGR+7%Met group showed diminished HHcy-induced erectile dysfunction(ED),indicating the improvement caused by hKLKl.Regarding corpus cavernosum endothelial cells,hKLKl preserved endothelial cell-cell junctions and endothelial cell content,and activated protein kinase B/endothelial nitric oxide synthase(Akt/eNOS)signaling.Fibrosis assessment indicated that hKLKl preserved normal penis structure by inhibiting apoptosis in the corpus cavernosum smooth muscle cells.Taken together,these findings showed that oxidative stress,impaired corpus cavernosum endothelial cells,and severe penis fibrosis were involved in the in duction of ED by HHcy in rats,whereas hKLKl preserved erectile function by inhibiting these pathophysiological changes.展开更多
目的探讨人源性激肽释放酶结合蛋白(kallistatin,KS)对椎间盘髓核细胞的作用及可能机制。方法在正常和退变髓核中检测KS的表达;过氧化氢/KS处理细胞后分为Control、H_(2)O_(2)、KS及H_(2)O_(2)+KS4组,检测各组细胞凋亡和细胞外基质(extr...目的探讨人源性激肽释放酶结合蛋白(kallistatin,KS)对椎间盘髓核细胞的作用及可能机制。方法在正常和退变髓核中检测KS的表达;过氧化氢/KS处理细胞后分为Control、H_(2)O_(2)、KS及H_(2)O_(2)+KS4组,检测各组细胞凋亡和细胞外基质(extracellular matrix,ECM)的变化;利用沉默信息调节因子2相关酶Ⅰ(silencing information regulator 2 related enzymeⅠ,SIRT1)小干扰RNA(siRNA-SIRT1)/KS处理细胞后分为Control、H_(2)O_(2)、KS+H_(2)O_(2)、KS+H_(2)O_(2)+siRNA-NC及KS+H_(2)O_(2)+siRNA-SIRT15组,检测各组髓核细胞凋亡及ECM的表达。结果退变椎间盘髓核中KS的mRNA和蛋白的表达水平明显低于正常髓核(P<0.05);相比于H_(2)O_(2)组,KS组及H_(2)O_(2)+KS组髓核细胞凋亡率明显降低(P<0.05),凋亡基因Caspase3、Caspase9和Bax mRNA及蛋白表达量明显降低(P<0.05),抗凋亡基因Bcl2表达量明显增加(P<0.05);GAG含量、CollagenⅡ和Aggrecan表达显著提高(P<0.05)。当siRNASIRT1敲降髓核细胞SIRT1表达后,相比于Control、KS+H_(2)O_(2)和KS+H_(2)O_(2)+siRNA-NC组,KS+H_(2)O_(2)+siRNA-SIRT1组凋亡率显著增加,GAG含量、CollagenⅡ和Aggrecan蛋白表达也显著降低(P<0.05)。结论KS可通过诱导SIRT1表达抑制H_(2)O_(2)诱导的人髓核细胞凋亡和ECM降解。展开更多
文摘The aim of this study was to investigate the mechanism by which a diet inducing high hyperhomocysteinemia(HHcy)leads to the deterioration of erectile function in rats and whether this is inhibited by expression of the human tissue kallikrein-1(hKLKl)gene.We established a rat model of HHcy by feeding methionine(Met)-rich diets to male Sprague-Dawley(SD)rats.Male wild-type SD rats(WTRs)and transgenic rats harboring the hKLKl gene(TGRs)were fed a normal diet until 10 weeks of age.Then,30 WTRs were randomly divided into three groups as follows:the control(n=10)group,the low-dose(4%Met,n=10)group,and the high-dose(7%Met,n=10)group.Another 10 age-matched TGRs were fed the high-dose diet and designated as the TGR+7%Met group.After 30 days,in all four groups,erectile function was measured and penile tissues were harvested to determine oxidative stress,endothelial cell content,and penis fibrosis.Compared with the 7%Met group,the TGR+7%Met group showed diminished HHcy-induced erectile dysfunction(ED),indicating the improvement caused by hKLKl.Regarding corpus cavernosum endothelial cells,hKLKl preserved endothelial cell-cell junctions and endothelial cell content,and activated protein kinase B/endothelial nitric oxide synthase(Akt/eNOS)signaling.Fibrosis assessment indicated that hKLKl preserved normal penis structure by inhibiting apoptosis in the corpus cavernosum smooth muscle cells.Taken together,these findings showed that oxidative stress,impaired corpus cavernosum endothelial cells,and severe penis fibrosis were involved in the in duction of ED by HHcy in rats,whereas hKLKl preserved erectile function by inhibiting these pathophysiological changes.
文摘目的探讨人源性激肽释放酶结合蛋白(kallistatin,KS)对椎间盘髓核细胞的作用及可能机制。方法在正常和退变髓核中检测KS的表达;过氧化氢/KS处理细胞后分为Control、H_(2)O_(2)、KS及H_(2)O_(2)+KS4组,检测各组细胞凋亡和细胞外基质(extracellular matrix,ECM)的变化;利用沉默信息调节因子2相关酶Ⅰ(silencing information regulator 2 related enzymeⅠ,SIRT1)小干扰RNA(siRNA-SIRT1)/KS处理细胞后分为Control、H_(2)O_(2)、KS+H_(2)O_(2)、KS+H_(2)O_(2)+siRNA-NC及KS+H_(2)O_(2)+siRNA-SIRT15组,检测各组髓核细胞凋亡及ECM的表达。结果退变椎间盘髓核中KS的mRNA和蛋白的表达水平明显低于正常髓核(P<0.05);相比于H_(2)O_(2)组,KS组及H_(2)O_(2)+KS组髓核细胞凋亡率明显降低(P<0.05),凋亡基因Caspase3、Caspase9和Bax mRNA及蛋白表达量明显降低(P<0.05),抗凋亡基因Bcl2表达量明显增加(P<0.05);GAG含量、CollagenⅡ和Aggrecan表达显著提高(P<0.05)。当siRNASIRT1敲降髓核细胞SIRT1表达后,相比于Control、KS+H_(2)O_(2)和KS+H_(2)O_(2)+siRNA-NC组,KS+H_(2)O_(2)+siRNA-SIRT1组凋亡率显著增加,GAG含量、CollagenⅡ和Aggrecan蛋白表达也显著降低(P<0.05)。结论KS可通过诱导SIRT1表达抑制H_(2)O_(2)诱导的人髓核细胞凋亡和ECM降解。
