Qihuang Jianpi Zishen Granules improves thrombocytopenia in mice with systemic lupus erythematosus by suppressing platelet autophagy via the Ca2+/CaMKK2/AMPK/mTOR signaling pathway
Objective To explore the mechanism of Qihuang Jianpi Zishen Granules(QJZG)for improving thrombocytopenia in a mouse model of systemic lupus erythematosus(SLE).Methods Twenty-four MRL/lpr lupus mice were randomized equally into 4 groups for treatment with daily gavage of saline,QJZG or prednisone(Pred)or intraperitoneal injection(twice a week)of CaMKK2 activator,with 6 C57BL/6 mice with saline gavage as the control group.After 8 weeks of treatment,the mice were examined for PLT,PCT,PDW,MPV,serum levels of TPO,IL-6,IL-10,TNF-α and IFN-γ,and calcium ion fluorescence intensity using ELISA or flow-through assay.RT-qPCR was used to detect platelet CaMKK2,AMPK2α,mTOR,Beclin1 and p62 mRNA expression levels,and the protein expressions of CaMKK2,p-CaMKK2,AMPK,p-AMPK,mTOR,p-mTOR,LC3,Beclin1 and p62 were detected using Western blotting.Results The saline-treated MRL/lpr lupus mice showed significantly lowered levels of PLT,PCT,IL-10,mTOR,p62 mRNA,p-mTOR and P62 with increased PDW,MPV,serum TPO,IL-6,TNF-α and IFN-γ levels,and platelet expressions of CaMKK2,AMPK,Bcl-1 mRNA,p-CaMKK2,p-AMPK,LC3Ⅱ and Beclin1.These abnormalities were significantly improved in QJZG group and Pred group but worsened after treatment with the CaMKK2 activator.Conclusion QJZG can ameliorate thrombocytopenia in mouse models of SLE by reducing inflammation and inhibiting platelet autophagy via regulating the Ca2+/CaMKK2/AMPK/mTOR signaling pathways.
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