楮实子预处理对对乙酰氨基酚致L02细胞损伤的保护作用  

Protective effect of fructus broussonetiae on APAP-induced injury of L02 cells

作　　者：韩世诚 王婷婷 张一昕 王茜 柴天川[2] HAN Shicheng;WANG Tingting;ZHANG Yixin;WANG Xi;CHAI Tianchuan(College of Pharmacy,Hebei University of Traditional Chinese Medicine,Hebei Province,Shijiazhuang,050091,China;Department of Pharmacy,Hebei Provincial Hospital of Traditional Chinese Medicine,Hebei Province,Shijiazhuang,050091,China)

机构地区：[1]河北中医药大学药学院,河北石家庄050091 [2]河北省中医院制剂科,河北石家庄050091

出　　处：《中国医药导报》2024年第1期34-38,共5页China Medical Herald

基　　金：河北省中医药管理局科研计划项目(2022362)。

摘　　要：目的探讨楮实子对对乙酰氨基酚(APAP)引起的L02细胞损伤的保护作用。方法CCK-8法测定不同浓度(0.1、0.3、1.0、3.0、10.0 mg/ml)楮实子水提物溶液对正常L02细胞的存活率。L02细胞APAP染毒后,取对数生长期的L02细胞,分为正常组、APAP组(40 mmol/L)、楮实子组(1 mg/ml),采用CCK-8法测定细胞存活率,采用DCFA-DH法测定细胞内活性氧(ROS)水平。Hoechst 33258染色观察细胞凋亡;罗丹明123染色检测L02细胞内线粒体膜电位表达情况。Western blot法检测细胞JNK、p-JNK、78GRP78、CHOP的表达水平。结果与0.1 mg/ml楮实子水提物比较,0.3、1.0、3.0、10.0 mg/ml楮实子水提物细胞活性降低,差异有统计学意义(P<0.01)。与正常组比较,APAP组细胞存活率下降,细胞凋亡平均荧光强度值升高(P<0.05);与APAP组比较,1 mg/ml楮实子水提物细胞存活率升高,细胞凋亡平均荧光强度值降低(P<0.05)。APAP组ROS荧光强度值高于正常组,差异有高度统计学意义(P<0.01);楮实子组ROS荧光强度值低于APAP组,差异有高度统计学意义(P<0.01)。APAP组线粒体膜电位平均荧光强度值低于正常组,差异有高度统计学意义(P<0.01);楮实子组线粒体膜电位平均荧光强度值高于APAP组,差异有高度统计学意义(P<0.01)。各组JNK蛋白表达水平比较,差异无统计学意义(P>0.05)。与正常组比较,APAP组p-JNK、GRP78、CHOP蛋白表达水平升高(P<0.01);与APAP组比较,楮实子组p-JNK、GRP78、CHOP蛋白表达水平降低(P<0.01)。结论楮实子可能通过抑制JNK活化,修复线粒体膜通透性,抑制内质网应激,减少ROS含量,抑制细胞凋亡,发挥保护APAP致L02细胞损伤的作用。Objective To investigate the protective effect of fructus broussonetiae on L02 cell damage induced by acetaminophen(APAP).Methods The survival rate of normal L02 cells was determined by CCK-8 method at different concentrations(0.1,0.3,1.0,3.0,10.0 mg/ml).After the L02 cells were infected with APAP,the L02 cells with logarithmic growth stage were selected and divided into normal group,APAP group(40 mmol/L)and fructus broussonetiae group(1 mg/ml).The cell survival rate was determined by CCK-8 method,and the intracellular ROS levels were determined by DCFA-DH method.Apoptosis was observed by Hoechst 33258 staining.The expression of mitochondrial membrane potential in L02 cells was detected by Rhodamine 123 staining.The expression levels of JNK,p-JNK,78GRP78,and CHOP were detected by Western blot.Results Compared with 0.1 mg/ml fructus broussonetia water extract,the cell activity of 0.3,1.0,3.0,and 10.0 mg/ml fructus broussonetia water extract decreased,and the difference was highly statistically significant(P<0.01).Compared with normal group,the cell survival rate and the average fluorescence intensity of apoptosis of APAP group were decreased(P<0.05).Compared with APAP group,the survival rate of 1 mg/ml fructus broussonetia water extract increased,and the average fluorescence intensity of apoptosis decreased(P<0.05).The ROS fluorescence inten-sity of APAP group was higher than that of normal group,and the differences were highly st atistically significant(P<0.01).The ROS fluorescence intensity of fructus broussonetiae seed group was lower than that of APAP group,and the differences were highly statistically significant(P<0.01).The average fluorescence intensity of mitochondrial membrane potential in APAP group was lower than that in normal group,and the difference was highly statistically significant(P<0.01).The average fluorescence intensity of mitochondrial membrane potential in fructus broussonetia group was higher than that in APAP group,and the difference was highly statistically significant(P<0.01).There was

关 键 词：对乙酰氨基酚 楮实子 药物性肝损伤 线粒体功能 内质网应激 

分 类 号：R965[医药卫生—药理学]