中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
46期
7447-7451
,共5页
姚丹丹%马瑞霞%翟丽慧%李作林%李臻
姚丹丹%馬瑞霞%翟麗慧%李作林%李臻
요단단%마서하%적려혜%리작림%리진
组织构建%组织工程%血管紧张素Ⅱ%足细胞%高糖%足细胞瞬时受体电位阳离子通道蛋白6(TRPC6)%nephrin%血管紧张素抑制剂%糖尿病肾病%蛋白尿%高渗%U73122%山东省自然科学基金
組織構建%組織工程%血管緊張素Ⅱ%足細胞%高糖%足細胞瞬時受體電位暘離子通道蛋白6(TRPC6)%nephrin%血管緊張素抑製劑%糖尿病腎病%蛋白尿%高滲%U73122%山東省自然科學基金
조직구건%조직공정%혈관긴장소Ⅱ%족세포%고당%족세포순시수체전위양리자통도단백6(TRPC6)%nephrin%혈관긴장소억제제%당뇨병신병%단백뇨%고삼%U73122%산동성자연과학기금
angiotensin II%podocytes%diabetes melitus%TRPC cation channels
背景：瞬时受体电位阳离子通道蛋白6是足细胞上一种新发现的阳离子通道蛋白，是组成裂隙隔膜重要蛋白之一，其表达变化与糖尿病肾病蛋白尿密切相关。目的：观察高糖刺激对足细胞瞬时受体电位阳离子通道蛋白6表达的影响，探讨糖尿病肾病发生发展的可能机制。方法：以永生化小鼠足细胞(MPC5)作为实验对象，将其设为：正常糖组(D-葡萄糖5.6 mmol/L)；渗透压对照组(D-葡萄糖5.6 mmol/L+甘露醇25 mmol/L)；高糖组(D-葡萄糖30 mmol/L)；缬沙坦组(D-葡萄糖30 mmol/L+10-5结果与结论：与正常糖组相比，高糖组足细胞瞬时受体电位阳离子通道蛋白6、血管紧张素Ⅱ蛋白及 mRNA水平明显升高(P<0.01)，nephrin mRNA及蛋白水平明显降低(P<0.01)；与高糖组相比，缬沙坦组瞬时受体电位阳离子通道蛋白6、血管紧张素Ⅱ的表达显著降低(P<0.05，P<0.01)；高糖+U73122组瞬时受体电位阳离子通道蛋白6、血管紧张素Ⅱ表达较高糖组明显下降(P<0.05，P<0.01)；渗透压对照组与正常糖组之间各因子差异无显著性意义(P>0.05)。结果说明高糖可能通过血管紧张素Ⅱ-瞬时受体电位阳离子通道蛋白6反馈信号通路损伤足细胞，同时也为血管紧张素受体拮抗剂通过此通路保护足细胞而治疗糖尿病肾病的提供了一新的理论基础。 mol/L缬沙坦)；高糖+U73122组(D-葡萄糖30 mmol/L+10μmol/L磷脂酶抑制剂U73122)。各组细胞干预时间为48 h。采用荧光定量PCR和western-blot方法检测各组足细胞瞬时受体电位阳离子通道蛋白6、nephrin、血管紧张素ⅡmRNA和蛋白的表达。
揹景：瞬時受體電位暘離子通道蛋白6是足細胞上一種新髮現的暘離子通道蛋白，是組成裂隙隔膜重要蛋白之一，其錶達變化與糖尿病腎病蛋白尿密切相關。目的：觀察高糖刺激對足細胞瞬時受體電位暘離子通道蛋白6錶達的影響，探討糖尿病腎病髮生髮展的可能機製。方法：以永生化小鼠足細胞(MPC5)作為實驗對象，將其設為：正常糖組(D-葡萄糖5.6 mmol/L)；滲透壓對照組(D-葡萄糖5.6 mmol/L+甘露醇25 mmol/L)；高糖組(D-葡萄糖30 mmol/L)；纈沙坦組(D-葡萄糖30 mmol/L+10-5結果與結論：與正常糖組相比，高糖組足細胞瞬時受體電位暘離子通道蛋白6、血管緊張素Ⅱ蛋白及 mRNA水平明顯升高(P<0.01)，nephrin mRNA及蛋白水平明顯降低(P<0.01)；與高糖組相比，纈沙坦組瞬時受體電位暘離子通道蛋白6、血管緊張素Ⅱ的錶達顯著降低(P<0.05，P<0.01)；高糖+U73122組瞬時受體電位暘離子通道蛋白6、血管緊張素Ⅱ錶達較高糖組明顯下降(P<0.05，P<0.01)；滲透壓對照組與正常糖組之間各因子差異無顯著性意義(P>0.05)。結果說明高糖可能通過血管緊張素Ⅱ-瞬時受體電位暘離子通道蛋白6反饋信號通路損傷足細胞，同時也為血管緊張素受體拮抗劑通過此通路保護足細胞而治療糖尿病腎病的提供瞭一新的理論基礎。 mol/L纈沙坦)；高糖+U73122組(D-葡萄糖30 mmol/L+10μmol/L燐脂酶抑製劑U73122)。各組細胞榦預時間為48 h。採用熒光定量PCR和western-blot方法檢測各組足細胞瞬時受體電位暘離子通道蛋白6、nephrin、血管緊張素ⅡmRNA和蛋白的錶達。
배경：순시수체전위양리자통도단백6시족세포상일충신발현적양리자통도단백，시조성렬극격막중요단백지일，기표체변화여당뇨병신병단백뇨밀절상관。목적：관찰고당자격대족세포순시수체전위양리자통도단백6표체적영향，탐토당뇨병신병발생발전적가능궤제。방법：이영생화소서족세포(MPC5)작위실험대상，장기설위：정상당조(D-포도당5.6 mmol/L)；삼투압대조조(D-포도당5.6 mmol/L+감로순25 mmol/L)；고당조(D-포도당30 mmol/L)；힐사탄조(D-포도당30 mmol/L+10-5결과여결론：여정상당조상비，고당조족세포순시수체전위양리자통도단백6、혈관긴장소Ⅱ단백급 mRNA수평명현승고(P<0.01)，nephrin mRNA급단백수평명현강저(P<0.01)；여고당조상비，힐사탄조순시수체전위양리자통도단백6、혈관긴장소Ⅱ적표체현저강저(P<0.05，P<0.01)；고당+U73122조순시수체전위양리자통도단백6、혈관긴장소Ⅱ표체교고당조명현하강(P<0.05，P<0.01)；삼투압대조조여정상당조지간각인자차이무현저성의의(P>0.05)。결과설명고당가능통과혈관긴장소Ⅱ-순시수체전위양리자통도단백6반궤신호통로손상족세포，동시야위혈관긴장소수체길항제통과차통로보호족세포이치료당뇨병신병적제공료일신적이론기출。 mol/L힐사탄)；고당+U73122조(D-포도당30 mmol/L+10μmol/L린지매억제제U73122)。각조세포간예시간위48 h。채용형광정량PCR화western-blot방법검측각조족세포순시수체전위양리자통도단백6、nephrin、혈관긴장소ⅡmRNA화단백적표체。
BACKGROUND:Transient receptor potential channel C6 (TRPC6) is a new and important slit diaphragm-associated protein in podocytes involved in regulating glomerular filter function. Glomerular TRPC6 expression is closely associated with proteinuria in diabetic kidney disease. OBJECTIVE: To investigate the expression of canonical TRPC6 in mouse podocytes induced by high glucose, and to explore the possible mechanism of diabetic kidney disease. METHODS:Mouse podocyte cels were cultured and divided into normal glucose group (5.6 mmol/L D-glucose), normal control group (5.6 mmol/L D-glucose+25 mmol/L mannitol) and experimental groups which were in the environment of high glucose (30 mmol/L). The experimental groups included high glucose group, valsartan treatment groups (10-5 mol/L) and U73122 control group (10μmol/L U73122). After 48 hours, the expressions of mRNA and proteins of TRPC6, nephrin and angiotensin II (AngII) were detected respectively by real-time quantitative PCR and western blot analysis. RESULTS AND CONCLUSION:Compared with the normal control group, the expressions of mRNA and proteins of TRPC6 and angiotensin II were markedly elevated in the high glucose group (P < 0.01), while the expressions of mRNA and proteins of nephrin were decreased (P < 0.01). The mRNA and proteins of TRPC6 and angiotensin II expressions were significantly down-regulated by valsartan (P < 0.05,P < 0.01), while the mRNA and protein expressions of nephrin were effectively up-regulated (P < 0.05). Compared with the high glucose group, the expressions of mRNA and proteins of TRPC6 and angiotensin II were ameliorated in the U73122 control group. The expressions of mRNA and proteins of TRPC6, nephrin and angiotensin II had no statistical significance between the normal control group and normal glucose group (P > 0.05). Angiotensin II-TRPC6 signaling pathway may mediate high glucose-induced podocyte injury, meanwhile it provides a new theoretical basis for the treatment of diabetic kidney disease, by which the angiotensin receptor blockers can protect podocytes in diabetic kidney disease.
