摘要
目的 探讨血管紧张素Ⅱ (AngⅡ )受体拮抗剂 (ARB)替米沙坦和血管紧张素转换酶抑制剂 (A CEI)苯那普利对负鼠近端小管上皮细胞 (OK细胞 )增殖和Na+ K+ ATP酶活性的影响。方法 培养的OK细胞采用低渗方法制备细胞膜悬液 ,以BCA 10 0蛋白质定量测定试剂盒测定膜蛋白 ;Na+ K+ ATP酶活性采用孔雀绿比色分析法测定释放的无机磷 (Pi)含量 ,培养液中分别加入AngⅡ、AngⅡ +替米沙坦、AngⅡ +苯那普利 ,观察其对OK细胞Na+ K+ ATP酶活性的影响。结果 ①对照组Na+ K+ ATP酶活性为 (0 .0 896± 0 .0 0 6 6 )μmol·mg蛋白质·L-1·h-1,1× 10 -10 mol/LAngⅡ组为 (0 .0 972± 0 .0 0 81) μmol·mg蛋白质·L-1·h-1,活性明显高于对照组 (P <0 .0 5 ) ;培养液中同时加入 1× 10 -10 mol/LAngⅡ和 1× 10 -9mol/L替米沙坦时为 (0 .0 6 2 3± 0 .0 0 5 3) μmol·mg蛋白质·L-1·h-1,较 1× 10 -10 mol/LAngⅡ组明显降低 (P <0 .0 5 ) ;培养液中同时加入1× 10 -10 mol/LAngⅡ和 1× 10 -9mol/L苯那普利为 (0 .10 2 7± 0 .0 0 17) μmol·mg蛋白质·L-1·h-1,与 1×10 -10 mol/LAngⅡ组的差异无显著性 (P >0 .0 5 )。②AngⅡ对OK细胞DNA合成有刺激作用 ,随着浓度的升高(1× 10 -10 ～ 1× 10 -6mol/L)作用?
Objective To appraise the effects of angiotensin Ⅱ type 1 receptor antagonist (Telmisartan) and angiotensin-converting enzyme inhibitor(Benazepril) on Na +-K +-ATPase activity and proliferation of tubular epithelium of kidney (OK) cell. Methods The cultured OK cell membranes were estimated by BCA-100 protein quantitation kit; Malachite green colorimetric analysis was used to detect the inorganic phosphorus released to determine the ouabain-sensitive Na +-K +-ATPase activity of OK cells. Adding AngⅡ, Telmisartan and benazepril respectively to the cultured fluid to observe the changes of proliferation of OK cells which could be detected by measuement of 3H-thymidine incorporation. Results ①AngⅡ (1×10 -10 mol/L) caused a significant rise in Na +-K +-ATPase activity [(0.097 2± 0.008 0) μmol·mg pro·L -1·h -1, vs (0.089 6±0.006 5) μmol·mg pro·L -1·h -1, P<0.05]. Telmisartan (1×10 -9 mol/L) reduced the increased activity by AngⅡ[(0.062 3±0.005 3) μmol·mg pro·L -1·h -1, vs (0.097 2±0.008 0) μmol·mg pro·L -1·h -1, P<0.05], whereas Benazepril (1×10 -9mol/L) did not affect Na +-K +-ATPase activity significantly [(0.102 7±0.016 6)vs (0.097 2±0.008 0) μmol·mg pro·L -1·h -1, P> 0.05]. ②The synthesis of DNA of opossum kidney cells was stimulated by AngⅡ, which was dose-dependent, but when the concentration of AngⅡ was raised to 1×10 -5 mol/L, on the contrary, the reaction decreased. ③The reduction in Angiotensin Ⅱ-stimulated thymidine incorporation by Telmisartan depended on the concentration at the range of 1×10 -6～1×10 -10 mol/L, with a maximal inhibition of 1×10 -6 mol/L by Telmisartan. ④The reduction in Angiotensin Ⅱ-stimulated thymidine incorporation by Benazepril was depended on the concentration at the range of 1×10 -6～1×10 -10 mol/L, with a maximal inhibition of 1×10 -6 mol/L by Benazepril. Conclusions ①AngⅡ increases Na +-K +-ATPase activity of OK cell. Telmisartan completely blocks the rise of Na +-K +-ATPase activity caused by Ang Ⅱ, whereas benazepril does not. Telmisartan can modulate the ion transport caused by AngⅡ. ②Angiotensin Ⅱ stimulats the growth of opossum kidney cells. Telmisartan and Angiotensin converting enzyme inhibitor benazepril can separately inhibit the cellular proliferation in response to stimulation by angiotensin Ⅱ in opossum kidney cells. Telmisartan and Benazepril have renal protective effect at cellular level.
出处
《上海医学》
CAS
CSCD
北大核心
2004年第6期389-391,共3页
Shanghai Medical Journal
