CAJ | 학술논문

目的研究周期素激酶抑制剂p27在高糖培养系膜细胞(MC)肥大中的作用.方法蛋白印迹(western杂交)方法测定MC裂解液p27蛋白水平,[3H]胸腺嘧啶核苷([3H]TdR)及[3H]亮氨酸([3H]leu)掺入方法测定MC细胞的肥大情况,观察p27反义寡核苷酸(ODN)转染对高糖培养MC肥大的影响.结果高糖(450mg/dl)无血清培养的MC同正常糖(100mg/dl)无血清培养的MC相比,p27水平增高,[3H]leu掺入增加,[3H]TdR掺入减少;p27反义ODN转染后高糖无血清培养MC[3H]leu掺入减少,[3H]TdR掺入增加;用甘露醇提高正常糖培养液渗透压并不增加MC中p27水平.结论高糖培养的MC中p27水平明显增高,增高的p27在高糖培养MC细胞肥大中起重要作用.
목적연구주기소격매억제제p27재고당배양계막세포(MC)비대중적작용.방법단백인적(western잡교)방법측정MC렬해액p27단백수평,[3H]흉선밀정핵감([3H]TdR)급[3H]량안산([3H]leu)참입방법측정MC세포적비대정황,관찰p27반의과핵감산(ODN)전염대고당배양MC비대적영향.결과고당(450mg/dl)무혈청배양적MC동정상당(100mg/dl)무혈청배양적MC상비,p27수평증고,[3H]leu참입증가,[3H]TdR참입감소;p27반의ODN전염후고당무혈청배양MC[3H]leu참입감소,[3H]TdR참입증가;용감로순제고정상당배양액삼투압병불증가MC중p27수평.결론고당배양적MC중p27수평명현증고,증고적p27재고당배양MC세포비대중기중요작용.
Objective Our study aimed at the role of p27 on the hypertrophy of mesangial cell (MC) cultured in high glucose. Methods The p27 protein of MC lysate was detected with western blotting analysis. The degree of cultured MC hypertrophy was estimated through [3H]-thymidine incorporation and [3H]-leucine incorporation. The effect of reducing p27 expression on cell hypertrophy was analysed with p27 antisense oligodeoxynucleotide (ODN) phosphorothioate. Results in MC cultured in high glucose (450mg/dl) serum-free DMEM compared with MC cultured in normal glucose (100mg/dl) serum-free DMEM, p27 increased, [3H]-leucine incorporation increased and[3H]-thymidine incorporation decreased;p27 antisense ODN transfection reduced [3H]leucine incorporation, increased [3H] thymidine incorporation of MC cultured in high glucose senumfree DMEM. Increasing medium osmolarity with D-mannitol failed to induce p27 expression of MC. Conclusion p27 protein increased in MC cultured in high glucose. High level of p27 played an important role in MC hypertrophy induced by high glucose. Because the cell cycle is controlled by the interaction between the positive cell cycle regulatory proteins(CCRP) and negative CCRP, further research is needed to study the expression of the positive and negative CCRP in MC in order to better understand the role of CCRP in MC hypertrophy.