蛋白激酶C-ε在尿蛋白所致肾小管上皮细胞损伤中的作用  

Effects of protein kinase C-ε on the injury of renal tubular epithelial cells induced by urinary proteins

作　　者：冯永民[1] 张媛媛[1] 杨陈[1] 陈孝文[1] 刘华锋[1] 

机构地区：[1]广东医学院附属医院肾病研究所,广东湛江524001

出　　处：《中国现代医学杂志》2013年第3期32-37,共6页China Journal of Modern Medicine

基　　金：广东省自然科学基金项目(No:8152402301000014);广东省高等学校高层次人才项目(No:SC1001)

摘　　要：目的探讨蛋白激酶Cε(PKC-ε)在尿蛋白所致肾小管上皮细胞(RTECs)损伤中的作用。方法将体外培养的人肾小管上皮细胞株(HK-2细胞)分为对照组、尿蛋白组(4 mg/mL),激动剂组(10μmol/LPKC-ε激动剂+4 mg/mL尿蛋白)及抑制剂组(10μmol/L PKC-ε抑制剂+4 mg/mL尿蛋白)分别处理48 h,用Western Blotting法检测PKC-ε活性和α平滑肌肌动蛋白(α-SMA)的表达;用ELISA方法检测培养上清人纤维连蛋白(FN)、转化生长因子β(TGF-β)的分泌水平;用全自动生化分析仪检测细胞LDH释放率。结果尿蛋白明显抑制HK-2细胞PKC-ε的活化(P<0.01),增加HK-2细胞α-SMA表达(P<0.05)、上清液FN和TGF-β分泌(P<0.01)以及LDH释放率(P<0.01);PKC-ε激动剂减轻尿蛋白所致HK-2细胞培养上清液TGF-β分泌(P<0.01),而不影响α-SMA表达、FN分泌以及LDH释放率;PKC-ε抑制剂进一步增加尿蛋白所致HK-2细胞α-SMA表达、FN和TGF-β分泌以及LDH释放率(均P<0.05)。结论 PKC-ε高活性是维持RTECs正常形态和功能的基础,尿蛋白抑制RTECs正常PKC-ε高活性,减轻尿蛋白对PKC-ε活性的抑制可能有利于维持RTECs正常的形态和功能。[ Objective] To investigate the role of PKC-ε in renal tubular epithelial cells （RTECs） injury induced by urinary protients. [ Methods ] Human renal tubular epithelial cell line （HK-2 cells） were divided into the follow- ing groups： control group, urinary protein group （4 mg/mL）, PKC-ε activator group （10 μmol/L PKC-ε activator＋4 mg/mL urinary protein） and PKC-ε inhibitor group （10 μmol/L PKC-ε inhibitor＋4 mg/mL urinary protein）. PKC-ε activity and α-smooth muscle actin （α-SMA） expression in HK-2 was detected by western blotting. The secretion of fibronectin （FN） and TGF-β in supernatant were determined by ELISA. The released rate of the lactic acid dehydro- genase （LDH） of HK-2 cells was measured by an automatic biochemistry analyzer. [Results] Urinary proteins strongly inhibited the activity of PKC-ε （P 〈0.01）, significantly promoted the expression of α-SMA （P 〈0.05）, secre- tion of FN and TGF-β （P 〈0.01）. Urinary proteins increased the release rates of LDH （P 〈0.01）. PKC-ε activator de- creased the secretion of TGF-β induced by urinary proteins （P 〈0.01）, while did not affected the expression of α- SMA, the secretion of FN and the release rates of LDH. PKC-ε inhibitor further increased the expression of SMA （P 〈0.05）, secretion of FN and TGF-I3 （P 〈0.05）. PKC-ε inhibitor further increased the release rates of LDH induced by urinary proteins （P 〈0.05）. [Conclusion] Normal high activity of PKC-ε is essential for maintaining the normalmorphology and function of RTECs. Activity of PKC-ε in RTECs can be inhibitated by urinary proteins. Reducing the inhibition on activity of PKC-ε in RTECs induced by urinary proteins may be useful to maintain the normal morphology and function of RTECs.

关 键 词：尿蛋白 肾小管上皮细胞 活性 蛋白激酶C-ε 

分 类 号：R696.2[医药卫生—泌尿科学]