葛根与粉葛抗H9c2细胞OGD/R损伤的有效成分及作用机制初步研究  

Preliminary Study on Differential Components and Mechanism of Puerariae Lobatae Radix and Puerariae Thomsonii Radix against OGD/R Injury in H9c2 Cells

作　　者：谢金根 刘荣华[1] 马嘉鑫 樊学程 雷婷 陈林林 肖雄[1] 高文军[1] 李文宏[1] XIE Jin-gen;LIU Rong-hua;MA Jia-xin;FAN Xue-cheng;LEI Ting;CHEN Lin-lin;XIAO Xiong;GAO Wen-jun;LI Wen-hong(Department of Pharmacy,Jiangxi University of Chinese Medicine,Nanchang 330004,China)

机构地区：[1]江西中医药大学药学院,南昌330004

出　　处：《中国药学杂志》2023年第3期243-253,共11页Chinese Pharmaceutical Journal

基　　金：国家重点研发计划项目资助(2017YFC1702902)。

摘　　要：目的初步探讨葛根(Puerariae Lobatae Radix,PLR)与粉葛(Puerariae Thomsonii Radix,PTR)抗心肌细胞损伤的物质基础、作用及机制的异同;比较二者不同含药体系中有效成分相对含量及抗心肌(H9c2)细胞氧-糖剥夺/复氧(oxygen-glucose deprivation/reoxygenation,OGD/R)损伤效应的差异。方法应用网络药理学方法找出PLR和PTR抗H9c2细胞OGD/R损伤的有效成分,结合超高效液相色谱-四极杆飞行时间串联质谱(UPLC-Q-TOF-MS)检测鉴定二者含药肠吸收液(intestinal absorption fluid,IAF)和含药血清(medicated serum,MS)中的主要有效成分及相对含量;建立H9c2细胞OGD/R损伤模型,PLR与PTR不同含药体系干预后,CCK-8检测细胞活力,生化试剂盒检测细胞内超氧化物歧化酶(superoxide dismutase,SOD)、乳酸脱氢酶(lactate dehydrogenase,LDH)活性,蛋白免疫印迹法(Western blot)检测细胞丝裂原活化蛋白激酶p38(MAPKp38)和核转录因子-κBp65(NFκBp65)蛋白表达。结果研究发现PLR和PTR抗H9c2细胞OGD/R损伤的有效成分为葛根素、滨蒿内酯、大豆素、芒柄花素、染料木素,且MS中各有效成分相对含量均远低于IAF;与正常组比较,模型组SOD活性降低(P<0.01),LDH活性升高(P<0.01),MAPKp38和NFκBp65蛋白表达增加(P<0.05);与模型组比较,各给药组均可以升高SOD活性(P<0.01),降低LDH活性(P<0.01),下调MAPKp38和NFκBp65蛋白表达(P<0.05)。比较发现,PLR抗H9c2细胞损伤的作用优于PTR;IAF对各药效指标的影响优于MS。结论PLR和PTR均可不同程度的保护OGD/R引起的H9c2细胞损伤,而PLR的作用较强,其作用机制可能是通过调控MAPKp38和NFκBp65蛋白表达发挥作用;IAF中有效成分相对含量较高,对各药效指标影响的较强而重现性较好,适合于中药体外药理学研究。OBJECTIVE To explore the similarities and differences of material basis,action,and mechanism of Puerariae Lobatae Radix(PLR)and Puerariae Thomsonii Radix(PTR)against myocardial cell injury,and compare the differences of relative amount of active ingredients and the effect against H9c2 cells OGD/R injury in different medicated systems.METHODS By means of network pharmacology,the active components of PLR and PTR against cardiomyocyte OGD/R injury were identified.The main active components and relative contents in medicated intestinal absorption fluid(IAF)and medicated serum(MS)of PLR and PTR were determined by UPLC-Q-TOF-MS.The OGD/R injury model in H9c2 cells was established and intervened with different drug-containing systems of PLR and PTR.The cell viability was detected by CCK-8,the intracellular superoxide dismutase(SOD)and lactate dehydrogenase(LDH)activities were detected by a biochemical kit,and the protein expressions of MAPKp38 and NFκBp65 in cells were detected by Western Blot.RESULTS The effective components of PLR and PTR against cardiomyocyte OGD/R injury were puerarin,scoparone,daidzein,formononetin,and genistein,and the relative content of each effective component in MS was much lower than that in IAF.Compared with the normal group,the SOD activity in the model group was decreased(P<0.01),the LDH activity was increased(P<0.01),and the protein expressions of MAPKp38 and NFκBp65 were increased(P<0.05).Compared with the model group,each administration group could increase SOD activity(P<0.01),reduce LDH activity(P<0.01),and down-regulate the expression of MAPKp38 and NFκBp65 proteins(P<0.05).The comparison showed that the anti-H9c2 cell injury effect of PLR was superior to that of PTR,and the effect of IAF on each pharmacodynamic index was better than that of MS.CONCLUSION PLR and PTR can protect H9c2 cells against injury induced by OGD/R in different degrees,while PLR playing a stronger role,which might be through the regulation of MAPKp38 and NFκBp65 protein expressions.The relative content of e

关 键 词：葛根 粉葛 含药肠吸收液 氧-糖剥夺/复氧 心肌细胞 

分 类 号：R966[医药卫生—药理学]