摘要
目的 探究梓醇(CAT)通过调节酪氨酸蛋白激酶2(JAK2)/信号转导及转录激活蛋白3(STAT3)信号通路对铅诱导的PC12细胞神经毒性的影响。方法 将PC12细胞随机分为空白组、模型组(25μmol·L^(-1)醋酸铅)、低、高剂量实验组(2.5、5.0 mg·L^(-1)CAT)、高剂量实验+激活剂组(5.0 mg·L^(-1)CAT+0.5μmol·L^(-1)colivelin)。用乳酸脱氢酶(LDH)实验检测细胞损伤,用流式细胞术检测细胞凋亡率,用荧光染色检测细胞内活性氧(ROS)水平,用实时荧光定量聚合酶链反应法测定细胞JAK2、STAT3 mRNA的表达情况,用蛋白质印迹法检测细胞中JAK2/STAT3信号通路和凋亡相关蛋白的表达情况。结果 空白组、模型组、高剂量实验组和高剂量实验+激活剂组的B细胞白血病/淋巴瘤抗原-2蛋白相对表达水平分别为0.80±0.08、0.33±0.03,0.74±0.07和0.45±0.04,LDH水平分别为(214.28±12.07)、(376.50±28.46)、(243.25±14.88)和(332.49±18.15)U·L^(-1),ROS相对荧光强度分别为1.00±0.00、1.89±0.19、1.20±0.12和1.75±0.17,凋亡率分别为(4.35±0.57)%、(26.59±1.96)%、(11.07±1.08)%和(21.63±2.06)%,JAK2 mRNA表达水平分别为1.00±0.00、1.86±0.18、1.28±0.13和1.30±0.13,STAT3 mRNA表达水平分别为1.00±0.00、1.94±0.19、1.34±0.13和1.72±0.17,p-JAK2/JAK2分别为0.26±0.03、0.78±0.08、0.38±0.04和0.36±0.07,p-STAT3/STAT3分别为0.23±0.02、0.84±0.08、0.42±0.04和0.75±0.07,Bax蛋白表达水平分别为0.31±0.03、0.86±0.08、0.40±0.04和0.69±0.07。模型组的上述指标与空白组比较,差异均有统计学意义(均P<0.05);高剂量实验组的上述指标与模型组比较,差异均有统计学意义(均P<0.05)。结论 CAT可能通过抑制JAK2/STAT3信号通路来减轻铅诱导的PC12细胞神经毒性。
Objective To investigate the impacts of catalpol(CAT)on lead-induced neurotoxicity of PC 12 cells by regulating Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signal pathway.Methods PC 12 cells were randomly grouped into blank group(blank medium treatment),model group(25μmol·L^(-1)lead acetate),experimental-L group(25μmol·L^(-1)lead acetate+2.5mg·L^(-1)CAT),experimental-H group(25μmol·L^(-1)lead acetate+5.0 mg·L^(-1)CAT)and experimental-H+colivelin group(25μmol·L^(-1)lead acetate+5.0 mg·L^(-1)CAT+0.5μmol·L^(-1)colivelin).Lactate dehydrogenase(LDH)assay was applied to detect cell damage;the apoptosis rate was detected by flow cytometry;the level of reactive oxygen species(ROS)was detected by fluorescence staining;the mRNA expression levels of JAK2 and STAT73 were measured by real-time quantitative polymerase chain reaction;Western blot was applied to detect the expression of JAK2/STAT3 signal pathway and apoptosis-related proteins in cells.Results The expression levels of B cell lymphomal/leukemia-2protein in the blank group,model group,experimental-H group,experimental-H+colivelin group were 0.80±0.08,0.33±0.03,0.74±0.07 and 0.45±0.04;the cell LDH levels were(214.28±12.07),(376.50±28.46),(243.25±14.88)and(332.49±18.15)U·L^(-1);the ROS relative fluorescence intensity were 1.00±0.00,1.89±0.19,1.20±0.12 and 1.75±0.17;the cell apoptosis rates were(4.35±0.57)%,(26.59±1.96)%,(11.07±1.08)%and(21.63±2.06)%;the expression levels of JAK2 mRNA were 1.00±0.00,1.86±0.18,1.28±0.13 and 1.30±0.13;the expression levels of STA73 mRNA were 1.00±0.00,1.94±0.19,1.34±0.13and 1.72±0.17;the phosphorylation levels of p-JAK2/JAK2 were 0.26±0.03,0.78±0.08,0.38±0.04 and0.36±0.07;the phosphorylation levels of p-STAT3/STAT3 were 0.23±0.02,0.84±0.08,0.42±0.04 and0.75±0.07;the expression levels of Bax protein in cells were 0.31±0.03,0.86±0.08,0.40±0.04 and0.69±0.07.The above indicators in the model group showed statistically significant differences compared to the
作者
王宁
鲁欣然
王磊
何佳起
王维展
WANG Ning;LU Xin-ran;WANG Lei;HE Jia-qi;WANG Wei-zhan(Department of Emergency Medicine,Harrison International Peace Hospital,Hebei Medical University,Hengshui 053000,Hebei Province,China;Department of Radiotherapy,Harrison International Peace Hospital,Hebei Medical University,Hengshui 053000,Hebei Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2023年第23期3419-3423,共5页
The Chinese Journal of Clinical Pharmacology
基金
衡水市科技计划基金资助项目(2018014002Z)。
