The expression of angiopoietin-1(Ang-1) and thrombospondin-1(TSP-1) in 5/6 subtotal nephrectomy(STN) rats model,and its correlation to the renal microvasculature injury were investigated.Rat 5/6 STN model was establis...The expression of angiopoietin-1(Ang-1) and thrombospondin-1(TSP-1) in 5/6 subtotal nephrectomy(STN) rats model,and its correlation to the renal microvasculature injury were investigated.Rat 5/6 STN model was established in adult male SD rats,and the shamoperated group and 5/6 STN group were set up.The renal function and histopathological changes were examined at the 1st,2nd,4th,8th and 12th week after operation.The expression of Ang-1,TSP-1 and CD31 in renal tissues was detected by using immunohistochemistry.From 2nd to 8th week after operation,Ang-1 was significantly expressed in glomeruli of rats with STN.Ang-1 staining in glomeruli of STN group was increased significantly as compared with that in sham-operated group at 4th and 8th week after operation,and subsequently decreased after the 12th week.The expression of TSP-1 was increased significantly in STN group.As compared with sham-operated group,the CD31 expression was significantly down-regulated from the 2nd week.The expression of Ang-1 mRNA was detected by using RT-PCR at the same time points.The expression of Ang-1 mRNA in renal tissue of rats with STN was significantly up-regulated at the 2nd,4th and 8th week after operation as compared with that in STN group at other time points or in sham-operated group at the same time points,while decreased evidently at the 12th week as compared with that in sham-operated group.It is concluded that there are changes in the mRNA expression of Ang-1,and the significant up-regulation of the expression of TSP-1 in renal tissue of rats with STN,which may be involved in the remnant renal microvasculature injury.展开更多
This study investigated the effects of benazepril administered in the morning or evening on the diurnal variation of renin-angiotensin-aldosterone system (RAAS) and clock genes in the kidney. The male Wistar rat model...This study investigated the effects of benazepril administered in the morning or evening on the diurnal variation of renin-angiotensin-aldosterone system (RAAS) and clock genes in the kidney. The male Wistar rat models of 5/6 subtotal nephrectomy (STNx) were established. Animals were randomly divided into 4 groups: sham STNx group (control), STNx group, morning benazepril group (MB) and evening benazepril group (EB). Benazepril was intragastrically administered at a dose of 10 mg/kg/day at 07:00 and 19:00 in the MB group and EB group respectively for 12 weeks. All the animals were synchronized to the light:dark cycle of 12:12 for 12 weeks. Systolic blood pressure (SBP), 24-h urinary protein excretion and renal function were measured at 11 weeks. Blood samples and kidneys were collected every 4 h throughout a day to detect the expression pattern of renin activity (RA), angiotensin Ⅱ (AngⅡ) and aldosterone (Ald) by radioimmunoassay (RIA) and the mRNA expression profile of clock genes (bmal1, dbp and per2) by real-time PCR at 12 weeks. Our results showed that no significant differences were noted in the SBP, 24-h urine protein excretion and renal function between the MB and EB groups. There were no significant differences in average Ald and RA content of a day between the MB group and EB group. The expression peak of bmal1 mRNA was phase-delayed by 4 to 8 h, and the diurnal variation of per2 and dbp mRNA diminished in the MB and EB groups compared with the control and STNx groups. It was concluded when the similar SBP reduction, RAAS inhibition and clock gene profile were achieved with optimal dose of benazepril, morning versus evening dosing of benazepril has the same renoprotection effects.展开更多
Aim Thromboxane A2（TXA2） is assumed to contribute to the process of renal dysfunction. The pres- ent study was designed to investigate whether terutroban, a specific antagonist of thromboxane/prostaglandin？ （TP） ...Aim Thromboxane A2（TXA2） is assumed to contribute to the process of renal dysfunction. The pres- ent study was designed to investigate whether terutroban, a specific antagonist of thromboxane/prostaglandin？ （TP） receptor, protects against renal damage in 5/6 nephrectomy. Methods C57/BL6 mice were randomly grouped into sham-operated （2K）, 5/6 nephroectomy groups （5/6K-off） and 5/6 nephroectomy treated with ter- utroban （10 mg · kg^-1 · d^-l） groups （Yerutroban）. Renal artery and kidney were collected for vascular function study, Western blot, immunohistochemistry （IHC） assay and enzyme-linked immunosorbent assay （ELISA）, re- spectively. Results Four weeks after the surgery, arterial blood pressure was comparable among the three groups. However mice in terutroban group had higher levels of serum creatinine and lower survival. Compared with 2K groups, 5/6K-off mice had significantly higher levels of renal blood flow as well as a blunted relaxation to acetyl- choline. Production of prostacyclin （PGI2） and thromboxane B2 （ TXB2）, but no prostaglandin E2 （ PGE2）, were significantly increased in the renal artery of 5/6K-off group. Terutroban restored the renal blood flow, but not the acetylcholine-induced relaxation in the renal artery. It is probably due to the blockade effect of terutroban on the smooth muscle since terutroban treatment significantly reduced U46619-induced vasconstriction in renal arteries. Interestingly, terutroban increased the production of TXB2, but not PGI2 or PGE2, in the renal artery. This proba-bly is a compensatory effect on prostaglandins production. In kidney cortex, 5/6K-off group had significantly lower levels of PGE2 and TXB2 when compared with 2 K group. Terutroban markedly increased all three prostaglandins levels. Conclusion Terutroban restores renal artery function, but not renal function in mouse with 5/6 nephrecto- my. It suggests that kidney has more complicated regulations than renal artery. High levels of prostanoids in kid- neys may contribute to renal damage in terutroban group. Further experiments will focus on examining the underly- ing mechanisms.展开更多
目的:研究溶质载体家族6成员9(solute carrier family 6 member 9,SLC6A9)表达对结直肠癌细胞增殖、迁移和5-氟尿嘧啶(5-fluorouracil,5-FU)药物敏感性的影响。方法:TCGA数据库分析、实时荧光定量PCR和Western blot分析检测SLC6A9在结...目的:研究溶质载体家族6成员9(solute carrier family 6 member 9,SLC6A9)表达对结直肠癌细胞增殖、迁移和5-氟尿嘧啶(5-fluorouracil,5-FU)药物敏感性的影响。方法:TCGA数据库分析、实时荧光定量PCR和Western blot分析检测SLC6A9在结肠癌组织、正常结肠细胞系(NCM460)和结直肠癌细胞系(SW620、HCT116、HT29、Lovo和SW480)中的表达。将SCL6A9过表达质粒及阴性对照(SLC6A9 OE、Vector)转染HT29细胞,将SCL6A9小干扰RNA及阴性对照(SLC6A9 siRNA1#、siRNA2#和Scramble)转染SW620细胞。划痕愈合实验和Transwell实验检测各组细胞的迁移、侵袭能力。Western blot和细胞免疫荧光检测EMT相关蛋白E-cadherin、Vimentin的表达水平。利用CCK-8法和构建裸鼠移植瘤模型检测SLC6A9过表达对结直肠癌细胞5-FU药物敏感性的影响。结果:与正常结肠组织和NCM460细胞相比,SLC6A9在结肠癌组织和结直肠癌细胞系中低表达(均P<0.05)。SLC6A9过表达引起E-cadherin蛋白表达增加,Vimentin蛋白水平降低,抑制结直肠癌细胞的迁移、侵袭(P<0.05)。SLC6A9低表达引起E-cadherin蛋白表达降低,Vimentin蛋白水平增加,促进结直肠癌细胞的迁移、侵袭能力(P<0.05)。SLC6A9过表达提高了5-FU的药物敏感性,并使肿瘤生长缓慢,质量减轻(P<0.05)。而SLC6A9低表达降低了5-FU的药物敏感性(P<0.05)。结论:SLC6A9过表达能够抑制结直肠癌细胞的迁移、侵袭和EMT进程,并增强5-FU对结直肠癌细胞的药物敏感性。展开更多
文摘The expression of angiopoietin-1(Ang-1) and thrombospondin-1(TSP-1) in 5/6 subtotal nephrectomy(STN) rats model,and its correlation to the renal microvasculature injury were investigated.Rat 5/6 STN model was established in adult male SD rats,and the shamoperated group and 5/6 STN group were set up.The renal function and histopathological changes were examined at the 1st,2nd,4th,8th and 12th week after operation.The expression of Ang-1,TSP-1 and CD31 in renal tissues was detected by using immunohistochemistry.From 2nd to 8th week after operation,Ang-1 was significantly expressed in glomeruli of rats with STN.Ang-1 staining in glomeruli of STN group was increased significantly as compared with that in sham-operated group at 4th and 8th week after operation,and subsequently decreased after the 12th week.The expression of TSP-1 was increased significantly in STN group.As compared with sham-operated group,the CD31 expression was significantly down-regulated from the 2nd week.The expression of Ang-1 mRNA was detected by using RT-PCR at the same time points.The expression of Ang-1 mRNA in renal tissue of rats with STN was significantly up-regulated at the 2nd,4th and 8th week after operation as compared with that in STN group at other time points or in sham-operated group at the same time points,while decreased evidently at the 12th week as compared with that in sham-operated group.It is concluded that there are changes in the mRNA expression of Ang-1,and the significant up-regulation of the expression of TSP-1 in renal tissue of rats with STN,which may be involved in the remnant renal microvasculature injury.
基金supported by grants from the Department of Public Health of Hubei Province of China (No. 2012Z-B08)the Health Bureau of Wuhan City of China (No. WX12C10)
文摘This study investigated the effects of benazepril administered in the morning or evening on the diurnal variation of renin-angiotensin-aldosterone system (RAAS) and clock genes in the kidney. The male Wistar rat models of 5/6 subtotal nephrectomy (STNx) were established. Animals were randomly divided into 4 groups: sham STNx group (control), STNx group, morning benazepril group (MB) and evening benazepril group (EB). Benazepril was intragastrically administered at a dose of 10 mg/kg/day at 07:00 and 19:00 in the MB group and EB group respectively for 12 weeks. All the animals were synchronized to the light:dark cycle of 12:12 for 12 weeks. Systolic blood pressure (SBP), 24-h urinary protein excretion and renal function were measured at 11 weeks. Blood samples and kidneys were collected every 4 h throughout a day to detect the expression pattern of renin activity (RA), angiotensin Ⅱ (AngⅡ) and aldosterone (Ald) by radioimmunoassay (RIA) and the mRNA expression profile of clock genes (bmal1, dbp and per2) by real-time PCR at 12 weeks. Our results showed that no significant differences were noted in the SBP, 24-h urine protein excretion and renal function between the MB and EB groups. There were no significant differences in average Ald and RA content of a day between the MB group and EB group. The expression peak of bmal1 mRNA was phase-delayed by 4 to 8 h, and the diurnal variation of per2 and dbp mRNA diminished in the MB and EB groups compared with the control and STNx groups. It was concluded when the similar SBP reduction, RAAS inhibition and clock gene profile were achieved with optimal dose of benazepril, morning versus evening dosing of benazepril has the same renoprotection effects.
文摘Aim Thromboxane A2（TXA2） is assumed to contribute to the process of renal dysfunction. The pres- ent study was designed to investigate whether terutroban, a specific antagonist of thromboxane/prostaglandin？ （TP） receptor, protects against renal damage in 5/6 nephrectomy. Methods C57/BL6 mice were randomly grouped into sham-operated （2K）, 5/6 nephroectomy groups （5/6K-off） and 5/6 nephroectomy treated with ter- utroban （10 mg · kg^-1 · d^-l） groups （Yerutroban）. Renal artery and kidney were collected for vascular function study, Western blot, immunohistochemistry （IHC） assay and enzyme-linked immunosorbent assay （ELISA）, re- spectively. Results Four weeks after the surgery, arterial blood pressure was comparable among the three groups. However mice in terutroban group had higher levels of serum creatinine and lower survival. Compared with 2K groups, 5/6K-off mice had significantly higher levels of renal blood flow as well as a blunted relaxation to acetyl- choline. Production of prostacyclin （PGI2） and thromboxane B2 （ TXB2）, but no prostaglandin E2 （ PGE2）, were significantly increased in the renal artery of 5/6K-off group. Terutroban restored the renal blood flow, but not the acetylcholine-induced relaxation in the renal artery. It is probably due to the blockade effect of terutroban on the smooth muscle since terutroban treatment significantly reduced U46619-induced vasconstriction in renal arteries. Interestingly, terutroban increased the production of TXB2, but not PGI2 or PGE2, in the renal artery. This proba-bly is a compensatory effect on prostaglandins production. In kidney cortex, 5/6K-off group had significantly lower levels of PGE2 and TXB2 when compared with 2 K group. Terutroban markedly increased all three prostaglandins levels. Conclusion Terutroban restores renal artery function, but not renal function in mouse with 5/6 nephrecto- my. It suggests that kidney has more complicated regulations than renal artery. High levels of prostanoids in kid- neys may contribute to renal damage in terutroban group. Further experiments will focus on examining the underly- ing mechanisms.
文摘目的:研究溶质载体家族6成员9(solute carrier family 6 member 9,SLC6A9)表达对结直肠癌细胞增殖、迁移和5-氟尿嘧啶(5-fluorouracil,5-FU)药物敏感性的影响。方法:TCGA数据库分析、实时荧光定量PCR和Western blot分析检测SLC6A9在结肠癌组织、正常结肠细胞系(NCM460)和结直肠癌细胞系(SW620、HCT116、HT29、Lovo和SW480)中的表达。将SCL6A9过表达质粒及阴性对照(SLC6A9 OE、Vector)转染HT29细胞,将SCL6A9小干扰RNA及阴性对照(SLC6A9 siRNA1#、siRNA2#和Scramble)转染SW620细胞。划痕愈合实验和Transwell实验检测各组细胞的迁移、侵袭能力。Western blot和细胞免疫荧光检测EMT相关蛋白E-cadherin、Vimentin的表达水平。利用CCK-8法和构建裸鼠移植瘤模型检测SLC6A9过表达对结直肠癌细胞5-FU药物敏感性的影响。结果:与正常结肠组织和NCM460细胞相比,SLC6A9在结肠癌组织和结直肠癌细胞系中低表达(均P<0.05)。SLC6A9过表达引起E-cadherin蛋白表达增加,Vimentin蛋白水平降低,抑制结直肠癌细胞的迁移、侵袭(P<0.05)。SLC6A9低表达引起E-cadherin蛋白表达降低,Vimentin蛋白水平增加,促进结直肠癌细胞的迁移、侵袭能力(P<0.05)。SLC6A9过表达提高了5-FU的药物敏感性,并使肿瘤生长缓慢,质量减轻(P<0.05)。而SLC6A9低表达降低了5-FU的药物敏感性(P<0.05)。结论:SLC6A9过表达能够抑制结直肠癌细胞的迁移、侵袭和EMT进程,并增强5-FU对结直肠癌细胞的药物敏感性。
