蛋白激酶C对胰岛素抵抗骨骼肌细胞的影响  

Influence of protein kinase C on insulin resistance in skeletal muscle cells

作　　者：侯士方[1] 孟馨[1] 相泓冰[1] 王涤非[1] 

机构地区：[1]中国医科大学附属第一医院内分泌科,辽宁沈阳110001

出　　处：《中国组织化学与细胞化学杂志》2011年第3期232-235,共4页Chinese Journal of Histochemistry and Cytochemistry

基　　金：辽宁省高等学校科研计划项目(2010225002);辽宁省教育厅科技基金(L2010626);辽宁省自然基金(20092113);辽宁省科技厅专项科研基金(2008934)

摘　　要：目的探讨蛋白激酶C(Protein Kinase C,PKC)在棕榈酸(Palmitic Acid,PA)诱导的骨骼肌细胞胰岛素抵抗(Isulin Resistance,IR)中的作用。方法免疫荧光鉴定原代大鼠骨骼肌细胞,氧化酶-过氧化物酶偶联法(GOD-POD法)检测培养液中葡萄糖浓度。设立对照组、棕榈酸组(PA组)、罗格列酮组(Rosiglitazone,Ros组),每组一分为二,分别加PKC抑制剂白屈莱红碱(Chelerythrine Chloride,CC)与正常培养液作用1h,Western Blot检测PKB及P-Ser473 PKB表达水平。结果 90%以上的细胞-αsarcometric actin免疫荧光染色呈阳性反应,表明培养的细胞为骨骼肌细胞;0.6mmol/L的PA作用24h可诱导骨骼肌细胞产生胰岛素抵抗;PA组与对照组相比P-Ser473 PKB水平显著降低,与本组未加CC相比显著升高。同时,罗格列酮组及本组加CC中P-Ser473PKB水平均高于PA组。结论在PA诱导的骨骼肌细胞IR方面PKC起重要作用,罗格列酮与PKC抑制剂CC均能改善PA引起的IR。Objective To study the role of protein kinase C in the mechanism of insulin resistance（IR） induced by palmitic acid in skeletal muscle cells.Methods The primary rat skeletal muscle cells were identified by immunofluorescence,and the glucose concentration was measured by GOD-POD method.Control group,palmitic acid group and rosiglitazone group were each divided into two subgroups,which received either,the PKC inhibitor chelerythrine chloride or common culture solution for one hour.Levels of protein kinase B and its phosphorylated form（P-Ser473 PKB） were determined by Western blot.Results Immunofluorescent staining showed that more than 90% of the cells were positively stained for α-sarcometric actin,which proved that the obtained cells were skeletal muscle cells.After 24 hours of treatment with 0.6mmol/L PA,the skeletal muscle cells could be induced for IR.The level of P-Ser473 PKB in the PA group decreased significantly compared to that of the control group and elevated significantly compared to that of the PA group without CC.Meanwhile,the level of P-Ser473 PKB in rosiglitazone and rosiglitazone with CC groups were significantly elevated compared to that of the PA group.Conclusion PKC plays an important role in insulin resistance induced by PA in skeletal muscle cells.Both rosiglitazone and chelerythrine chloride can significantly improve IR induced by PA.

关 键 词：胰岛素抵抗 蛋白激酶C 蛋白激酶B 棕榈酸 骨骼肌细胞 

分 类 号：R322.57[医药卫生—人体解剖和组织胚胎学]