title: Effects of puerarin on intracellular Ca2+and cell viability of MDCK renal tubular cells.
abstract: Puerarin is a natural compound and has been used as herb medication in a number of countries, especially in Asia. The effect of puerarin on Ca2+signaling is unknown in renal cells. This study examined whether puerarin affected Ca2+physiology in MDCK renal tubular cells. Cytosolic free Ca2+levels ([Ca2+]i) were measured using the fluorescent dye fura-2. Cell viability was examined by using WST-1 assay. Puerarin induced [Ca2+]irises and the response was reduced by removing extracellular Ca2+. Puerarin-induced Ca2+entry was not altered by protein kinase C (PKC) activity, but was inhibited by nifedipine. In Ca2+-free medium, treatment with the endoplasmic reticulum Ca2+pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) or thapsigargin partly inhibited puerarin-evoked [Ca2+]irises. Inhibition of phospholipase C (PLC) with U73122 did not change puerarin-induced [Ca2+]irises. Puerarin at 25-50μM caused cytotoxicity, which was not reversed by pretreatment with the Ca2+chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). Collectively, in MDCK cells, puerarin induced [Ca2+]irises by evoking PLC-independent Ca2+release from the endoplasmic reticulum and other unknown stores, and Ca2+entry via nifedipine-sensitive, PKC-insensitive Ca2+entry pathways. Puerarin also caused Ca2+-independent cell death.
authors.affiliation: Department of Medicine, Chang Bing Show Chwan Memorial Hospital, Changhua County 50544, Taiwan.
authors.forename: He-Hsiung
authors.initials: HH
authors.lastname: Cheng
authors.affiliation: Department of Nursing, Division of Basic Medical Sciences, Chang Gung University of Science and Technology, Chia-Yi 61363, Taiwan; Chronic Diseases and Health Promotion Research Center, Chang Gung University of Science and Technology, Chia-Yi 61363, Taiwan.
authors.forename: Chiang-Ting
authors.initials: CT
authors.lastname: Chou
authors.affiliation: Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan.
authors.forename: Wei-Zhe
authors.initials: WZ
authors.lastname: Liang
authors.affiliation: Department of Nursing, Tzu Hui Institute of Technology, Pingtung 92641, Taiwan.
authors.forename: Chun-Chi
authors.initials: CC
authors.lastname: Kuo
authors.affiliation: Department of Pharmacy, Tajen University, Pingtung 90741, Taiwan. Electronic address: pshieh@vghks.gov.tw.
authors.forename: Pochuan
authors.initials: P
authors.lastname: Shieh
authors.affiliation: Department of Rehabilitation, Kaohsiung Veterans General Hospital Tainan Branch, Tainan 71051, Taiwan. Electronic address: Jlwang@vghks.gov.tw.
authors.forename: Jue-Long
authors.initials: JL
authors.lastname: Wang
authors.affiliation: Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan. Electronic address: crjan@isca.vghks.gov.tw.
authors.forename: Chung-Ren
authors.initials: CR
authors.lastname: Jan
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D004285:Dogs; D007529:Isoflavones / Q000633:toxicity*; D007684:Kidney Tubules / Q000166:cytology; D061985:Madin Darby Canine Kidney Cells
chemical_list: D007529:Isoflavones; D002118:Calcium; C033607:puerarin
keywords: Ca(2+); MDCK renal tubular cells; Nifedipine; Puerarin