金钱草总黄酮对草酸钙结晶肾损伤的作用机制  

The Mechanism of Total Flavonoids of Lysimachiae Herba on Kidney Injury Induced by Calcium Oxalate Crystallization

作　　者：解海杰[1] 高宏伟[1] 王军[1] 李传波 张昌文[1] 刘春雨[1] 

机构地区：[1]天津医科大学第二医院尿石症治疗中心,天津300211

出　　处：《中国中西医结合外科杂志》2018年第1期58-63,共6页Chinese Journal of Surgery of Integrated Traditional and Western Medicine

基　　金：天津市卫生和计划生育委员会中医中西医结合科研课题(2015134)

摘　　要：目的:探讨金钱草总黄酮对一水合草酸钙(COM)诱导肾小管上皮细胞凋亡所致肾损伤的影响及其机制。方法:应用不同浓度COM处理肾小管上皮细胞(HK-2)建立体外泌尿系结石细胞模型,将肾小管上皮细胞分为对照组、金钱草总黄酮处理组(TF组)、P38丝裂原活化蛋白激酶(p_38/MAPK)抑制组(SB组)、COM处理组(COM组)、COM和金钱草总黄酮处理组(CTF组)、COM和p_38/MAPK抑制组(CSB组)6组,WST-1法检测细胞存活率,Western blot检测人肾损伤分子(KIM-1)、(p-p)_38、p_38、自噬相关蛋白LC3-Ⅱ的表达,流式细胞术检测不同处理组的细胞凋亡数。结果:COM浓度为0.5、1、2、5 mmol/L时,细胞存活率分别为(85.02±5.50)%、(70.27±3.78)%、(58.09±3.23)%、(42.56±4.13)%,较对照组(99.83±3.65)%降低(P<0.05)。在COM处理的肾小管上皮细胞中,随着时间的增加,KIM-1、(p-p)_38及LC3-Ⅱ表达增加。COM可诱导HK-2细胞过度自噬和凋亡,CSB组细胞凋亡率较COM组降低[(7.91±0.70)%vs(29.90±1.07)%,P<0.05]。CSB组LC3-Ⅱ表达较COM组降低[(0.52±0.04)vs(1.17±0.04),P<0.05]。CTF组(p-p)_38蛋白水平较COM组降低[(0.66±0.06)vs(1.01±0.06),P<0.05]。随着金钱草总黄酮浓度的增加,HK-2细胞存活率也逐渐增加。CTF组细胞凋亡率较COM组降低[(10.54±1.07)%vs(29.90±1.07)%,P<0.05]。CTF组LC3-Ⅱ表达较COM组降低[(0.82±0.10)vs(1.17±0.04),P<0.05]。结论:金钱草总黄酮可通过阻断p_38/MAPK通路抑制肾小管上皮细胞的过度自噬和凋亡,减少肾小管上皮细胞的损伤,从而抑制泌尿系结石的形成。Objective To discuss the effect of total ？avonoids of Lysimachiae Herba （TFLH） on kidney injury induced by calcium oxalate monohydrate （COM）. Methods Human kidney proximal tubular epithelial cell line （HK-2 cells） was treated with different concentration of COM to simulate urinary calculi cell model in vitro. Then, cells were divided into six groups, namely control group, TFLH treated group （TF group）, p38/mitogen-activated protein kinase （MAPK） inhibitor group （SB group）, COM treated group （COM group）, COM and TFLH treated group （CTF group） and COM and p38/MAPK inhibitor group （CSB group）. WST-1 assay was used to measure cell viability. Kidney injury molecule-1 （KIM-1）, p-p38, p38 and cell autophagy related proteins LC3-Ⅱ were tested by Western blotting. Flow cytometry was used to detect the number of apoptotic cells. Results With the increase of COM concentration （0.5, 1, 2 and 5 mmol/L）, cell viability [（85.02±5.50）%, （70.27±3.78）%, （58.09±3.23）% and （42.56±4.13）%] was significantly decreased compared with control group （99.83±3.65）% （P 〈 0.05）. In renal tubular epithelial cells treated with COM, the expression of KIM-1, p-p38 and LC3-II were increased with the increase of time. COM could induce excessiveautophagy and apoptosis in HK-2 cells, and the apoptosis rate in CSB group was lower than that in COM group [（7.91±0.70）% vs （29.90±1.07）%, P 〈 0.05]. LC3-II expression in CSB group was lower than that in COM group [（0.52±0.04） vs（1.17±0.04）, P 〈 0.05]. The level of p-p38 decreased in CTF group compared with COM group [（0.66±0.06） vs （1.01±0.06）, P 〈 0.05]. With the increase of TFLH concentration, the survival rate of HK-2 cells increased gradually. The rate of apoptosis in CTF group was lower than that in COM group [（10.54±1.07）% vs （29.90±1.07）%, P 〈 0.05]. The expression of LC3-II in CTF group was lower than that in COM group [（0.82±0.10） vs （1.17±0.04）, P 〈 0.0

关 键 词：金钱草总黄酮 -水合草酸钙 泌尿系结石模型 P38丝裂原活化蛋白激酶 肾损伤 

分 类 号：R692[医药卫生—泌尿科学]