CaMKKβ Is Involved in AMP-Activated Protein Kinase Activation by Baicalin in LKB1 Deficient Cell Lines 英文参考文献.docVIP

CaMKKbIsInvolvedinAMP-ActivatedProteinKinase ActivationbyBaicalininLKB1DeficientCellLines YingMa.,FuzhenYang.,YingWang,ZhiyanDu,DaihuaLiu,HongxiaGuo,JingkangShen,HongliPeng* StateKeyLaboratoryofDrugResearch,ShanghaiInstituteofMateriaMedica,ChineseAcademyofSciences,Shanghai,PRChina Abstract AMP-activatedproteinkinase(AMPK)playsanimportantroleinmediatingenergymetabolismandiscontrolledmainlyby two upstream kinases, LKB1 or Ca /calmodulin-dependent protein kinase kinase-b (CaMKKb). Previously, we found that 2+ baicalin, one of the major flavonoids in a traditional Chinese herb medicine, Scutellaria baicalensis, protects against the development of hepatic steatosis in rats feeding with a high-fat diet by the activation of AMPK, but, the underlying mechanismforAMPKactivationisunknown.HereweshowthatintwoLKB1-deficientcells,HeLaandA549cells,baicalin activates AMPK by a Thr-172 phosphorylation and subsequent phosphorylation of its downstream target, acetyl CoA carboxylase,atSer-79,toasimilardegreeasdoesinHepG2cells(thatexpressLKB1).PharmacologicinhibitionofCaMKKb by its selective inhibitor STO-609 markedly inhibits baicalin-induced AMPK activation in both HeLa and HepG2 cells, indicatingthatCaMKKbistheresponsibleAMPKkinase.Wealsoshowthattreatmentofbaicalincausesalargerincreasein 2+ 2+ 2+ intracellularCa concentration([Ca ]),althoughthemaximallevelof[Ca ]iislowerinHepG2cellscomparedtoHeLa i cells.ChelationofintracellularfreeCa by EDTAandEGTA, ordepletionofintracellularCa storesbytheendoplasmic 2+ 2+ 2+ reticulumCa -ATPaseinhibitorthapsigarginabrogatesbaicalin-inducedactivationofAMPKinHeLacells.Neithercellular ATPnortheproductionofreactiveoxygenspeciesisalteredbybaicalin.Finally,inHeLacells,baicalintreatmentnolonger decreases intracellular lipid accumulation caused by oleic acid after inhibition of CaMKKb by STO-609. These results demonstrate that a potential Ca /CaMKKb dependent pathway is involved in the activation of AMPK by baicalin and suggestthatCaMKKblik