摘要
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（１００ ｍｇ／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］ ｌｅｎ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。
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.
出处
《中国糖尿病杂志》
CAS
CSCD
2001年第4期241-244,共4页
Chinese Journal of Diabetes
