LY333531, a PKCβ inhibitor, attenuates glomerular endothelial cell apoptosis in the early stage of mouse diabetic nephropathy via down-regulating swiprosin-1  

作　　者：Zhi-bin WANG Su ZHANG Ya LI Rong-mei WANG Ling-chang TONG Yue WANG Wei-ye LIU Ding-feng SU Ye TU Li-chao ZHANG Ling LI 

机构地区：[1]Department of Pharmacology, College of Pharmacy, Second Military Medical University, Shanghai 200433, China [2]Department of Pharmacy, Ningxia Medical University, Yinchuan 750021, China [3]Department of Pharmacy, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai 200071, China [4]Medical Affairs Department, Shanghai East Hospital, Tongji University, Shanghai 200120, China

出　　处：《Acta Pharmacologica Sinica》2017年第7期1009-1023,共15页中国药理学报（英文版）

基　　金：We thank Tong-hui MA （Dalian Medical University, China） and Wei-heng SU （Jilin University, China） for constructing the swiprosin-1 knockout mouse. This study was supported by grants from the National Natural Science Foundation of China （No_ 81273504, 81473258 and 81402941）, the Shanghai Municipal Commission of Health and Family Planning （No 20144Y0204 and 201540294）, the Science and Technology Commission of Shanghai Municipality （No 15140904600）, and the Natural Science Foundation of Shanghai （No 16ZR1434400）.

摘　　要：Glomerular endothelial cell （GEC） injury plays an important role in the early stage of diabetic nephropathy （DN）. Previous studies show that a PKCβ inhibitor is effective for treating DN. In the current study we further explored the effects and molecular mechanisms of PKCI3 inhibitors on GEC apoptosis in DN in streptozotocin-induced diabetic mice in vivo and high glucoseor PMA-treated human renal glomerular endothelial cells （HRGECs） in vitro. In the diabetic mice, hyperglycemia caused aggravated nephropathy and GEC apoptosis accompanied by significantly increased expression of swiprosin-1, a potentally pro-apoptotic protein. Administration of LY333531 （1 mg.kg-1.d-1 for 8 weeks） significantly attenuated both GEC apoptosis and swiprosin-1 upregulation in the diabetic mice. Similar results were observed in high glucoseor PMA-treated HRGECs in vitro. The pro-apoptotic role of swiprosin-1 was further examined using HRGECs treated with lentivirus mediating RNA interference or over-expression and swiprosin-1-knockout mice. Over-expression of swiprosin-1 in HRGECs resulted in increases in apoptosis and in caspase-9, caspase-3 and Bax expression. In contrast, knockdown of swiprosin-1 attenuated high glucoseor PMA-induced HRGECs apoptosis. Furthermore, over-expression of swiprosin-1 promoted interaction between swiprosin-1 and caspase-9 and increased the formation of apoptosomes. In diabetic swiprosin-1-/- mice, the kidney/body weight, urinary albumin, glomerular hypertrophy, mitochondrial apoptotic-associated proteins and GEC apoptosis were significantly attenuated as compared with those in diabetic swiprosin-1＋/＋ mice. These results demonstrate that swiprosin-1 is up-regulated by PKCβ in the early stage of DN, and that PKCβ facilitates GEC apoptosis through the mitochondrial-dependent pathway.Glomerular endothelial cell （GEC） injury plays an important role in the early stage of diabetic nephropathy （DN）. Previous studies show that a PKCβ inhibitor is effective for treating DN. In the current study we further explored the effects and molecular mechanisms of PKCI3 inhibitors on GEC apoptosis in DN in streptozotocin-induced diabetic mice in vivo and high glucoseor PMA-treated human renal glomerular endothelial cells （HRGECs） in vitro. In the diabetic mice, hyperglycemia caused aggravated nephropathy and GEC apoptosis accompanied by significantly increased expression of swiprosin-1, a potentally pro-apoptotic protein. Administration of LY333531 （1 mg.kg-1.d-1 for 8 weeks） significantly attenuated both GEC apoptosis and swiprosin-1 upregulation in the diabetic mice. Similar results were observed in high glucoseor PMA-treated HRGECs in vitro. The pro-apoptotic role of swiprosin-1 was further examined using HRGECs treated with lentivirus mediating RNA interference or over-expression and swiprosin-1-knockout mice. Over-expression of swiprosin-1 in HRGECs resulted in increases in apoptosis and in caspase-9, caspase-3 and Bax expression. In contrast, knockdown of swiprosin-1 attenuated high glucoseor PMA-induced HRGECs apoptosis. Furthermore, over-expression of swiprosin-1 promoted interaction between swiprosin-1 and caspase-9 and increased the formation of apoptosomes. In diabetic swiprosin-1-/- mice, the kidney/body weight, urinary albumin, glomerular hypertrophy, mitochondrial apoptotic-associated proteins and GEC apoptosis were significantly attenuated as compared with those in diabetic swiprosin-1＋/＋ mice. These results demonstrate that swiprosin-1 is up-regulated by PKCβ in the early stage of DN, and that PKCβ facilitates GEC apoptosis through the mitochondrial-dependent pathway.

关 键 词：LY333531 diabetic nephropathy glomerular endothelial cells PKC[3 PMA swiprosin-1 apoptosis 

分 类 号：Q255[生物学—细胞生物学] O658[理学—分析化学]