盐酸药根碱抗异丙肾上腺素诱导的心肌H9c2细胞肥大作用研究

Anti isoprenaline-induced cardiac hypertrophy effect of Jatrorrhizine hydrochloride on H9c2 cells

关键词:  盐酸药根碱  心肌肥大  异丙肾上腺素  钙离子浓度  心房钠肽  脑钠肽  β1肾上腺素受体

Key words:  jatrorrhizine hydrochloride  cardiac hypertrophy  isoprenaline  Ca²⁺  ANP  BNP  β1 adrenal receptor

摘要:

[目的]研究盐酸药根碱对异丙肾上腺素诱导的H9c2心肌细胞肥大的影响。[方法]采用体外培养的心肌H9c2细胞，通过检测细胞总蛋白的方法评价细胞肥大程度，采用实时聚合酶链反应（PCR）的方法检测盐酸药根碱对异丙肾上腺素诱导的心钠肽（ANP）和脑钠肽（BNP）mRNA水平上升的影响。使用钙-4试剂盒检测细胞内钙离子浓度。细胞转染β1肾上腺素受体，考察盐酸药根碱与β1肾上腺素受体竞争性结合能力。采用Western blot方法考察盐酸药根碱对异丙肾上腺素（ISO）诱导的ERK通路磷酸化的影响。[结果]盐酸药根碱显着抑制异丙肾上腺素诱导的H9c2细胞肥大。盐酸药根碱显着抑制异丙肾上腺素诱导的H9c2细胞中ANP及BNP mRNA 水平上升；ISO及盐酸药根碱与β1肾上腺素受体均具有一定的结合能力。盐酸药根碱抑制ISO诱导的ERK信号通路激活。[结论]盐酸药根碱能通过与β1肾上腺素受体结合发挥抑制ERK信号通路激活而抗心肌肥大作用。

Abstract:

[Objective] To investigate the effect of jatrorrhizine hydrochloride on isoprenaline (ISO) induced cardiac hypertrophy in H9c2 cells. [Methods] H9c2 cells were cultured in vitro. Using the quantity of total proteins in cells to evaluate the level of cardiac hypertrophy; Real-time PCR was used to investigate the effect of jatrorrhizine hydrochloride on increase of ANP and BNP mRNA induced by ISO. Calcium-4 kit was used to detect the concentration of Ca²⁺ in cells. After transfect β1 adrenal receptor into cells, the binding abilities of jatrorrhizine hydrochloride and ISO to β1 adrenal receptor were examined. Western blot was used to detect the effect of jatrorrhizine hydrochloride on activation of ERK pathway. [Results] Jatrorrhizine hydrochloride significantly inhibited H9c2 cell hypertrophy induced by ISO. Jatrorrhizine hydrochloride significantly inhibited the increase of ANP and BNP mRNA. Jatrorrhizine hydrochloride showed milder bind ability to β1 adrenal receptor compared with ISO. Jatrorrhizine hydrochloride significantly inhibited ISO induced ERK pathway activation. [Conclusion] Jatrorrhizine hydrochloride can inhibit ISO induced cardiac hypertrophy by binding to β1 adrenal receptor.