title: Puerarin induces mitochondria-dependent apoptosis in hypoxic human pulmonary arterial smooth muscle cells.
abstract: BACKGROUND
  Pulmonary vascular medial hypertrophy in hypoxic pulmonary arterial hypertension (PAH) is caused in part by decreased apoptosis in pulmonary artery smooth muscle cells (PASMCs). Puerarin, an isoflavone purified from the Chinese medicinal herb kudzu, ameliorates chronic hypoxic PAH in animal models. Here we investigated the effects of puerarin on apoptosis of hypoxic human PASMCs (HPASMCs), and to determine the possible underlying mechanisms.
  
  
  RESULTS
  HPASMCs were cultured for 24 h in normoxia or hypoxia (5% O₂) conditions with and without puerarin. Cell number and viability were determined with a hemacytometer or a cell counting kit. Apoptosis was detected with a TUNEL test, rhodamine-123 (R-123) fluorescence, a colorimetric assay, western blots, immunohistochemical staining and RT-PCR. Hypoxia inhibited mitochondria-dependent apoptosis and promoted HPASMC growth. In contrast, after puerarin (50 µM or more) intervention, cell growth was inhibited and apoptosis was observed. Puerarin-induced apoptosis in hypoxic HPASMCs was accompanied by reduced mitochondrial membrane potential, cytochrome c release from the mitochondria, caspase-9 activation, and Bcl-2 down-regulation with concurrent Bax up-regulation.
  
  
  CONCLUSIONS
  Puerarin promoted apoptosis in hypoxic HPASMCs by acting on the mitochondria-dependent pathway. These results suggest a new mechanism of puerarin relevant to the management of clinical hypoxic pulmonary hypertension.
authors.affiliation: Department of Geriatric Medicine, The First Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang, China.
authors.forename: Chan
authors.initials: C
authors.lastname: Chen
authors.forename: Chun
authors.initials: C
authors.lastname: Chen
authors.forename: Zhiyi
authors.initials: Z
authors.lastname: Wang
authors.forename: Liangxing
authors.initials: L
authors.lastname: Wang
authors.forename: Lehe
authors.initials: L
authors.lastname: Yang
authors.forename: Minjiao
authors.initials: M
authors.lastname: Ding
authors.forename: Cheng
authors.initials: C
authors.lastname: Ding
authors.forename: Yu
authors.initials: Y
authors.lastname: Sun
authors.forename: Quan
authors.initials: Q
authors.lastname: Lin
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Huang
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Du
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Zhao
authors.forename: Chuangyi
authors.initials: C
authors.lastname: Wang
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D001483:Base Sequence; D017931:DNA Primers; D006801:Humans; D000860:Hypoxia / Q000473:pathology*; D007529:Isoflavones / Q000494:pharmacology*; D008928:Mitochondria / Q000187:drug effects*; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000473:pathology; D011651:Pulmonary Artery / Q000187:drug effects* / Q000473:pathology; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D007529:Isoflavones; C033607:puerarin