四逆散及醋炙品组方含药血清对CORT致PC12细胞损伤的保护作用研究  

Investigation on Protective Effects of Medicated Sera of Sinisan Containing Bupleuri Radix and Paeoniae Radix Alba before and after Vinegar-Processing on CORT-Induced PC12 Cell Injury

作　　者：刘静[1,2] 蔡皓[1,2] 段煜[1,2] 裴科[3] 周佳 张雅婷[1,2] 莫子晴 钮敏洁 LIU Jing;CAI Hao;DUAN Yu;PEI Ke;ZHOU Jia;ZHANG Ya-ting;MO Zi-qing;NIU Min-jie(School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023, China;Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing, 210023, China;Institute of Pharmaceutical and Food Engineering, Shanxi University of Traditional Chinese Medicine, Taiyuan, 030024, China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]南京中医药大学国家教育部中药炮制规范化及标准化工程研究中心,江苏南京210023 [3]山西中医药大学制药与食品工程学院,山西太原030024

出　　处：《南京中医药大学学报》2020年第3期339-345,共7页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：国家自然科学基金(81673600);江苏省研究生科研创新计划(KYCX18_1637)。

摘　　要：目的探讨四逆散(SNS)及醋炙品组方四逆散(VPSNS)含药血清对皮质酮(CORT)致PC12细胞损伤的保护作用和抗抑郁作用的分子机制。方法采用MTT和形态观察法,确立模型的建立是否成功;经培养后的细胞分为4组:空白对照组、CORT组、SNS血清组、VPSNS血清组,采用ELISA法测定细胞上清液中多巴胺(DA)、5-羟色胺(5-HT)、肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)含量;采用微板法测定细胞上清液中乳酸脱氢酶(LDH)含量和超氧化物歧化酶(SOD)活力,并采用流式细胞术检测细胞凋亡率;采用Western blot和qPCR法检测细胞中BDNF、ERK和CREB蛋白和基因表达。结果与模型组相比,20%VPSNS血清组与20%SNS血清组均能显著提高DA、5-HT含量和SOD活力,以及BDNF、ERK和CREB蛋白和基因表达(P<0.05),并均能降低TNF-α、IL-1β和LDH含量以及细胞凋亡率(P<0.05)。与20%SNS血清组相比,20%VPSNS血清组对CORT致PC12细胞损伤的保护作用更佳(P<0.05)。结论BDNF-ERK-CREB通路可能参与了SNS和VPSNS对CORT致PC12细胞损伤的保护作用,且VPSNS的保护作用优于SNS。OBJECTIVE To investigate the molecular mechanism of anti depression and protective effects of medicated sera of Sinisan containing Bupleuri Radix and Paeoniae Radix Alba before and after vinegar processing on CORT induced PC12 cell injury.METHEODS Firstly,the MTT and morphological observation methods were used to evaluate whether the model was established successfully.Secondly,the cultured cells were divided into four groups(blank control group,CORT group,SNS serum group,VPSNS serum group).ELISA method was used to determine the contents of the monoamine neurotransmitters(DA and 5 HT)and the inflammatory cytokines(TNFαand IL 1β)in cell supernatant.Then,the content of the injury index(LDH)and the activity of the protective index(SOD)in cell supernatant were measured using the microplate method,and the cell apoptosis rates were measured by flow cytometric analysis.Finally,the protein and gene expression of BDNF,ERK,and CREB were detected by Western blot and qPCR.RESULTS Compared to the model group,both 20%SNS serum group and 20%VPSNS serum group increased the contents of DA,5 HT,the activity of SOD,and the protein and gene expression of BDNF,ERK,and CREB(P<0.05).And both of them reduced the contents of TNFα,IL 1β,LDH,and the cell apoptosis rates(P<0.05).The protective effect of 20%VPSNS serum group on CORT induced PC12 cell injury was better than that of 20%SNS serum group(P<0.05).CONCLUSION The BDNF ERK CREB pathway is probably involved in the protective effects of SNS and VPSNS on CORT induced PC12 cell injury,and the protective effect of VPSNS is better than that of SNS.

关 键 词：四逆散 炮制 皮质酮 PC12细胞 抗抑郁 

分 类 号：R285.5[医药卫生—中药学]