CAJ | 학술논문

目的探讨血管紧张素1a型受体(AT1aR)基因敲除小鼠肾脏局部肾素-血管紧张素系统(RAS)的组分改变对糖尿病肾病(DN)肾小球硬化的影响及其可能机制.方法 AT1aR基因敲除小鼠和野生型小鼠腹腔注射链脲佐菌素(STZ,300 mg/kg)诱导糖尿病模型12周后,取肾脏组织作冰冻组织切片,用激光捕获微切割技术分离肾小球,提取RNA.用实时定量PCR的方法检测肾小球内AT1aR、血管紧张素1b型受体(AT1bR)、血管紧张素2型受体(AT2R)、血管紧张素原、血管紧张素转化酶(ACE)、肾素、醛固酮合成酶(CYP11B2)的mRNA表达.PAS染色观察肾脏病理变化.免疫组化检测转化生长因子β1(TGF-β1)、1型纤溶酶原激活物抑制物(PAI-1)、单核细胞趋化因子1(MCP-1)和肾素的表达.比较不同基因型小鼠肾小球细胞外基质和各细胞因子的表达变化.结果与野生型小鼠相比,AT1aR基因敲除小鼠肾小球内AT1bR、血管紧张索原、肾素、CYP11B2的表达明显上调(P<0.05),AT2R表达下调,ACE无明显改变;AT1aR基因敲除小鼠肾小球细胞外基质明显增加(P<0.05),TGF-β1、PAI-1、MCP-1和肾素的表达均明显增加(P<0.05).结论 AT1aR基因敲除并不能使糖尿病小鼠肾脏病变改善.RAS组分的表达改变(AT1bR的上调和AT2的下调,肾素的上调和CYP11B2的上调)参与糖尿病肾小球病变过程.
목적 탐토혈관긴장소1a형수체(AT1aR)기인고제소서신장국부신소-혈관긴장소계통(RAS)적조분개변대당뇨병신병(DN)신소구경화적영향급기가능궤제.방법 AT1aR기인고제소서화야생형소서복강주사련뇨좌균소(STZ,300 mg/kg)유도당뇨병모형12주후,취신장조직작빙동조직절편,용격광포획미절할기술분리신소구,제취RNA.용실시정량PCR적방법검측신소구내AT1aR、혈관긴장소1b형수체(AT1bR)、혈관긴장소2형수체(AT2R)、혈관긴장소원、혈관긴장소전화매(ACE)、신소、철고동합성매(CYP11B2)적mRNA표체.PAS염색관찰신장병리변화.면역조화검측전화생장인자β1(TGF-β1)、1형섬용매원격활물억제물(PAI-1)、단핵세포추화인자1(MCP-1)화신소적표체.비교불동기인형소서신소구세포외기질화각세포인자적표체변화.결과 여야생형소서상비,AT1aR기인고제소서신소구내AT1bR、혈관긴장색원、신소、CYP11B2적표체명현상조(P<0.05),AT2R표체하조,ACE무명현개변;AT1aR기인고제소서신소구세포외기질명현증가(P<0.05),TGF-β1、PAI-1、MCP-1화신소적표체균명현증가(P<0.05).결론 AT1aR기인고제병불능사당뇨병소서신장병변개선.RAS조분적표체개변(AT1bR적상조화AT2적하조,신소적상조화CYP11B2적상조)삼여당뇨병신소구병변과정.
Objective To explore the glomerular change of renin-angiotensin system (RAS) expression in ATIaR gene knockout mice and its effects on extracellular matrix (ECM) remodeling under diabetic condition. Methods ATlaR knockout mice were generated previously. Hyperglycemia was induced by peritoneal injection of streptozotocin in ATIaR knockout mice and wild type mice. Normal AT1aR knockout mice and wild type mice were used as control group. Twelve weeks later, kidneys were harvested and frozen quickly in dry ice-acetone. Glomendi were collected by laser capture microdissection and total RNA was extracted, mRNA expression of AT1aR, AT1bR, AT2R, angiotensinogen, ACE, renin, and CYP11B2 was assessed by real-time PCR. ECM accumulation was evaluated by PAS staining. Protein levels of transforming growth factor β1(TGF-β1), type 1 plasminogen activator inhibitor(PAI-1), monocyte chemotactie protein 1(MCP-1) and renin were semi-quantitated by immunostaining. Results Compared to the wild type, mRNA expression of AT1bR, angiotensinogen, renin, CYP11B2 within glomeruli was upregulated significantly in ATlaR knockout mice (P<0.05), but no change of ACE expression was found in these two groups. AT2R protein was poorly detected in AT1aR knockout glomeruli and downregulated in wild type glomemli. ECM accumulation was significanfly increased associated with the parallel increase in TGF-β1, PAI-1, MCP-1 and renin within glomendi (P <0.05). Conclusions AT1aR gene knockout cannot improve ECM deposition in diabetic nephropathy. The compensate change of RAS components may be involved in this scenario: upregulation of AT1bR, downregulation of AT2R. CYP11B2 and renin may function in a novel pathway.