The renin angiotensin system(RAS)appears to influence male fertility at multiple levels.In this work,we analyzed the relationship between the RAS and DNA integrity.Fifty male volunteers were divided into two groups(25...The renin angiotensin system(RAS)appears to influence male fertility at multiple levels.In this work,we analyzed the relationship between the RAS and DNA integrity.Fifty male volunteers were divided into two groups(25 each):control(DNA fragmentation≤20%)and pathological(DNA fragmentation>20%)cases.Activities of five peptidases controlling RAS were measured fluorometrically:prolyl endopeptidase(which converts angiotensin[A]I and A II to A 1–7),neutral endopeptidase(NEP/CD10:A I to A 1–7),aminopeptidase N(APN/CD13:A III to A IV),aminopeptidase A(A II to A III)and aminopeptidase B(A III to A IV).Angiotensin-converting enzyme(A I to A II),APN/CD13 and NEP/CD10 were also assessed by semiquantitative cytometry and quantitative flow cytometry assays,as were the receptors of all RAS components:A II receptor type 1(AT1R),A II receptor type 2(AT2R),A IV receptor(AT4R or insulin-regulated aminopeptidase[IRAP]),(pro)renin receptor(PRR)and A 1–7 receptor or Mas receptor(MasR)None of the enzymes that regulate levels of RAS components,except for APN/CD13(decrease in fragmented cells),showed significant differences between both groups.Micrographs of RAS receptors revealed no significant differences in immunolabeling patterns between normozoospermic and fragmented cells.Labeling of AT1R(94.3%normozoospermic vs 84.1%fragmented),AT4R(96.2%vs 95.3%)and MasR(97.4%vs 87.2%)was similar between the groups.AT2R(87.4%normozoospermic vs 63.1%fragmented)and PRR(96.4%vs 48.2%)were higher in non-fragmented spermatozoa.These findings suggest that fragmented DNA spermatozoa have a lower capacity to respond to bioactive RAS peptides.展开更多
AIM: To evaluate the role of chymase in blood pressure regulation and its actions on tissue renin-angiotensin system.METHODS: A two-kidney, one-clip(2K1C) hypertension model was developed in Syrian hamsters, which hav...AIM: To evaluate the role of chymase in blood pressure regulation and its actions on tissue renin-angiotensin system.METHODS: A two-kidney, one-clip(2K1C) hypertension model was developed in Syrian hamsters, which have a human-type chymase. Either an angiotensin(Ang) converting enzyme(ACE) inhibitor(ACE-I; temocapril, 30 mg/kg per day), Ang Ⅱ type 1 receptor antagonist(ARB; CS866, 10 mg/kg per day), or vehicle was administered, beginning 2 wk after renal artery clipping and continued for 16 wk. At the end of this protocol, hearts, aortas, and lungs were removed, and total Ang Ⅱ-forming activities and ACE- and chymasedependent Ang Ⅱ-forming activities were determined.RESULTS: After renal artery clipping, systolic blood pressure in the vehicle group was significantly higher compared with that in a sham-operated group throughoutthe experimental period. Both ACE-I and ARB treatments revealed similar antihypertensive effects. Moreover, in the vehicle group, cardiac total and chymase-dependent Ang Ⅱ-forming activities significantly increased at 18 wk after clipping. Further, cardiac total and chymase-dependent Ang Ⅱ-forming activities decreased significantly after ACE-I or ARB treatment for 16 wk. In addition, chymase-dependent Ang Ⅱ-forming activity significantly increased in the aorta, although these changes were inhibited only by ARB. ARB treatment was more effective compared with ACE-I treatment in reversing the changes in tissue Ang Ⅱ formation, particularly in the aorta, despite their similar antihypertensive effects.CONCLUSION: Chymase does not play a major role in maintaining blood pressure and tissue ACE and chymase are regulated in a tissue-dependent manner in 2K1 C hamster.展开更多
为阐明失重致大动脉区域特异性重塑的特点和重力性对抗措施的机理,我们进行了整体与血管培养实验研究。整体实验:以大鼠尾部悬吊法模拟失重4周(SUS组),以1h/d恢复站立体位模拟间断性人工重力(S+D组),并设同步对照组(CON)。观察模拟失重...为阐明失重致大动脉区域特异性重塑的特点和重力性对抗措施的机理,我们进行了整体与血管培养实验研究。整体实验:以大鼠尾部悬吊法模拟失重4周(SUS组),以1h/d恢复站立体位模拟间断性人工重力(S+D组),并设同步对照组(CON)。观察模拟失重对大鼠大动脉血管(颈总动脉和腹主动脉)的重塑影响及对抗措施的效果;用免疫组织化学、蛋白免疫印迹分析、原位杂交及实时PCR等方法检测血管组织血管紧张素原(angiotensinogen,Ao)和血管紧张素II1型受体(angiotensin II type 1 receptor,AT1R)的蛋白与基因表达变化,以揭示血管局部肾素-血管紧张素系统(local renin-angiotensin system,L-RAS)是否参与调节。结果显示:与CON组相比,SUS组颈总动脉与腹主动脉近管腔的平滑肌层分别发生肥厚与萎缩变化;Ao与AT1R表达分别上调和下调。但在S+D组,上述适应性变化可被完全防止。血管培养实验:本文建立了可以控制流量、压力的血管培养系统,并得到下述结果。在高压(150mmHg)下培养颈总动脉3d,可引起平滑肌层c-纤维粘连蛋白表达增强,且以靠近管腔的肌层最为显著;培养液Ang II的生成量也增多;但如在3d灌流期间,每天有4或1h使灌流压降至0或80mmHg,则上述改变即可被完全防止。以上结果支持我们的假说:跨壁压分布变化是失重引起动脉区域性重塑的始动原因;但每日只要短时间使其恢复常态,血管重塑及L-RAS的变化即可被完全防止。展开更多
基金This work was supported by grants from the University of the Basque Country(UPV/EHU GIU 17/19)the Gangoiti Barrera Foundation(Basque Country).
文摘The renin angiotensin system(RAS)appears to influence male fertility at multiple levels.In this work,we analyzed the relationship between the RAS and DNA integrity.Fifty male volunteers were divided into two groups(25 each):control(DNA fragmentation≤20%)and pathological(DNA fragmentation>20%)cases.Activities of five peptidases controlling RAS were measured fluorometrically:prolyl endopeptidase(which converts angiotensin[A]I and A II to A 1–7),neutral endopeptidase(NEP/CD10:A I to A 1–7),aminopeptidase N(APN/CD13:A III to A IV),aminopeptidase A(A II to A III)and aminopeptidase B(A III to A IV).Angiotensin-converting enzyme(A I to A II),APN/CD13 and NEP/CD10 were also assessed by semiquantitative cytometry and quantitative flow cytometry assays,as were the receptors of all RAS components:A II receptor type 1(AT1R),A II receptor type 2(AT2R),A IV receptor(AT4R or insulin-regulated aminopeptidase[IRAP]),(pro)renin receptor(PRR)and A 1–7 receptor or Mas receptor(MasR)None of the enzymes that regulate levels of RAS components,except for APN/CD13(decrease in fragmented cells),showed significant differences between both groups.Micrographs of RAS receptors revealed no significant differences in immunolabeling patterns between normozoospermic and fragmented cells.Labeling of AT1R(94.3%normozoospermic vs 84.1%fragmented),AT4R(96.2%vs 95.3%)and MasR(97.4%vs 87.2%)was similar between the groups.AT2R(87.4%normozoospermic vs 63.1%fragmented)and PRR(96.4%vs 48.2%)were higher in non-fragmented spermatozoa.These findings suggest that fragmented DNA spermatozoa have a lower capacity to respond to bioactive RAS peptides.
文摘AIM: To evaluate the role of chymase in blood pressure regulation and its actions on tissue renin-angiotensin system.METHODS: A two-kidney, one-clip(2K1C) hypertension model was developed in Syrian hamsters, which have a human-type chymase. Either an angiotensin(Ang) converting enzyme(ACE) inhibitor(ACE-I; temocapril, 30 mg/kg per day), Ang Ⅱ type 1 receptor antagonist(ARB; CS866, 10 mg/kg per day), or vehicle was administered, beginning 2 wk after renal artery clipping and continued for 16 wk. At the end of this protocol, hearts, aortas, and lungs were removed, and total Ang Ⅱ-forming activities and ACE- and chymasedependent Ang Ⅱ-forming activities were determined.RESULTS: After renal artery clipping, systolic blood pressure in the vehicle group was significantly higher compared with that in a sham-operated group throughoutthe experimental period. Both ACE-I and ARB treatments revealed similar antihypertensive effects. Moreover, in the vehicle group, cardiac total and chymase-dependent Ang Ⅱ-forming activities significantly increased at 18 wk after clipping. Further, cardiac total and chymase-dependent Ang Ⅱ-forming activities decreased significantly after ACE-I or ARB treatment for 16 wk. In addition, chymase-dependent Ang Ⅱ-forming activity significantly increased in the aorta, although these changes were inhibited only by ARB. ARB treatment was more effective compared with ACE-I treatment in reversing the changes in tissue Ang Ⅱ formation, particularly in the aorta, despite their similar antihypertensive effects.CONCLUSION: Chymase does not play a major role in maintaining blood pressure and tissue ACE and chymase are regulated in a tissue-dependent manner in 2K1 C hamster.
基金supported by the grants from the National Natural Science Foundation of China(No.3047064930800545+2 种基金31070839)US NIH(HL095258)NSF(CAREER award 0644646)
文摘为阐明失重致大动脉区域特异性重塑的特点和重力性对抗措施的机理,我们进行了整体与血管培养实验研究。整体实验:以大鼠尾部悬吊法模拟失重4周(SUS组),以1h/d恢复站立体位模拟间断性人工重力(S+D组),并设同步对照组(CON)。观察模拟失重对大鼠大动脉血管(颈总动脉和腹主动脉)的重塑影响及对抗措施的效果;用免疫组织化学、蛋白免疫印迹分析、原位杂交及实时PCR等方法检测血管组织血管紧张素原(angiotensinogen,Ao)和血管紧张素II1型受体(angiotensin II type 1 receptor,AT1R)的蛋白与基因表达变化,以揭示血管局部肾素-血管紧张素系统(local renin-angiotensin system,L-RAS)是否参与调节。结果显示:与CON组相比,SUS组颈总动脉与腹主动脉近管腔的平滑肌层分别发生肥厚与萎缩变化;Ao与AT1R表达分别上调和下调。但在S+D组,上述适应性变化可被完全防止。血管培养实验:本文建立了可以控制流量、压力的血管培养系统,并得到下述结果。在高压(150mmHg)下培养颈总动脉3d,可引起平滑肌层c-纤维粘连蛋白表达增强,且以靠近管腔的肌层最为显著;培养液Ang II的生成量也增多;但如在3d灌流期间,每天有4或1h使灌流压降至0或80mmHg,则上述改变即可被完全防止。以上结果支持我们的假说:跨壁压分布变化是失重引起动脉区域性重塑的始动原因;但每日只要短时间使其恢复常态,血管重塑及L-RAS的变化即可被完全防止。
