健脾滋肾方通过PI3K/AKT通路对MRL/Lpr狼疮鼠血小板凋亡的影响  

Effect of Jianpi Zishen formula on platelet cell apoptosis in MRL/Lpr lupus rats through PI3K/AKT pathway

作　　者：汤忠富 黄传兵[1] 尚双双 陈君洁 程丽丽 李明[1] 刘思娣 TANG Zhong-fu;HUANG Chuan-bing;SHANG Shuang-shuang;CHEN Jun-jie;CHEN Li-li;LI Ming;LIU Si-di(The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,Anhui,China)

机构地区：[1]安徽中医药大学第一附属医院,安徽合肥230031

出　　处：《时珍国医国药》2024年第14期3127-3133,共7页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(82104782);安徽省省级中医优势专科建设项目(皖中医药服务秘[2022]34号);安徽省高校协同创新项目(GXXT-2021-085);安徽省临床医学研究转化专项项目(20230429510702014)。

摘　　要：目的探讨健脾滋肾方对MRL/Lpr狼疮鼠PI3K/AKT血小板凋亡通路的影响。方法将C57BL/6小鼠作为空白组,将MRL/lpr狼疮小鼠随机分为模型组、健脾滋肾方低剂量组(6.32g/kg)、健脾滋肾方高剂量组(12.64g/kg)、泼尼松组(5mg/kg),每组6只。检测小鼠外周血中血小板计数(PLT)、抗双链DNA抗体(anti-dsDNA)、抗核抗体(ANA)、补体3(C3)水平;RT-qPCR检测PI3K/AKT通路相关因子表达量;蛋白质免疫印迹法检测PI3K/AKT通路相关因子和凋亡因子Bax、Bcl-2、Caspase-3表达;流式细胞术检测血小板细胞凋亡情况。结果与空白组比较,模型组PLT、C3、Bcl-2 mRNA、Bcl-2蛋白显著降低(P<0.01);anti-dsDNA、ANA显著升高(P<0.01);PI3K mRNA、AKT mRNA、Bax mRNA相对表达量显著升高(P<0.01);P-PI3K、P-AKT、Bax、Caspase-3蛋白水平及血小板细胞凋亡率显著升高(P<0.01)。与模型组比较,健脾滋肾方低剂量组、高剂量组及泼尼松组PLT、C3、Bcl-2 mRNA、Bcl-2均显著升高(P<0.01,P<0.05),anti-dsDNA、ANA显著降低(P<0.01),PI3K、AKT、Bax mRNA相对表达量显著降低(P<0.01),P-PI3K、P-AKT、Bax、Caspase-3蛋白表达显著降低(P<0.01),血小板凋亡率均显著降低(P<0.01)。结论健脾滋肾方可能通过PI3K/AKT通路,调节血小板细胞凋亡,从而改善SLE血小板减少。Objective To investigate the effect of Jianpi Zishen formula on PI3K/AKT platelet apoptosis pathway in MRL/Lpr lupus rats.Methods C57BL/6 mice were taken as blank group,and MRL/lpr lupus mice were randomly divided into model group,Jianpi Zishen recipe low-dose group(6.32g/kg),Jianpi Zishen recipe high-dose group(12.64g/kg)and prednisone group(5mg/kg),with 6 mice in each group.Platelet count(PLT),anti-dsDNA,anti-nuclear antibody(ANA)and complement 3(C3)levels in peripheral blood of mice were detected.The expression of related factors in PI3K/AKT pathway was detected by RT-qPCR.The expressions of PI3K/AKT pathway related factors and apoptosis factors Bax,Bcl-2 and Caspase-3 were detected by Western blot.The apoptosis of platelet cells was detected by flow cytometry.Results Compared with blank group,PLT,C3,Bcl-2 mRNA and Bcl-2 protein in model group were significantly decreased(P<0.01).anti-dsDNA and ANA were significantly increased(P<0.01).The relative expressions of PI3K mRNA,AKT mRNA and Bax mRNA were significantly increased(P<0.01).The protein levels of P-PI3K,P-Akt,Bax,Caspase-3 and the apoptosis rate of platelet cells were significantly increased(P<0.01).Compared with model group,PLT,C3,Bcl-2 mRNA and Bcl-2 were significantly increased in Jianpi Zishen recipe low-dose group,high-dose group and prednisone group(P<0.01,P<0.05),while anti-dsDNA and ANA were significantly decreased(P<0.01).The relative mRNA expression levels of PI3K,AKT and Bax were significantly decreased(P<0.01),the protein expressions of P-PI3K,P-Akt,Bax and Caspase-3 were significantly decreased(P<0.01),and the apoptosis rate of platelet was significantly decreased(P<0.01).Conclusion Jianpi Zishen formula may regulate the apoptosis of platelet cells through PI3K/AKT pathway,so as to ameliorate SLE thrombocytopenia.

关 键 词：健脾滋肾方 系统性红斑狼疮 PI3K/AKT 血小板凋亡 

分 类 号：R285.5[医药卫生—中药学]