Abacavir, nevirapine, and ritonavir modulate intracellular calcium levels without affecting GHRH-mediated growth hormone secretion in somatotropic cells in vitro
Abacavir, nevirapine, and ritonavir modulate intracellular calcium levels without affecting GHRH-mediated growth hormone secretion in somatotropic cells in vitro
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DOI：
10.1016/j.mce.2018.12.005
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发表时间：
2019-02-15
影响因子：
4.1
通讯作者：
Casarini, Livio
中科院分区：
文献类型：
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作者：
Brigante, Giulia;Riccetti, Laura;Casarini, Livio
在接受抗逆转录病毒治疗治疗的HIV感染患者中，生长激素（GH）缺乏症经常出现。我们用抗逆转录病毒（Nevirapine，Ritonavir或Abacavir硫酸盐； 100 pm-1 mm范围）治疗了GH3细胞，并在人类生长激素释放激素（GHRH）受体cDNA转染后治疗。细胞活力，细胞内cAMP，CREB的磷酸化和钙的增加，GH的产生和分泌在基础状态下以及GHRH之后，使用MTT，生物发光共振能量转移，Western blotting和Elisa.antirortoviral治疗不会影响GHRH 50％有效剂量（EC50）计算30分钟的细胞内营地增加（Mann-Whitney的U检验； P> = 0.05; n = 4）和15分钟的CREB磷酸化。 GHRH介导的快速细胞内钙的动力学因与药物的预孵育而受到干扰，而GHRH未能诱导利托纳维尔预处理细胞的离子增加（ANOVA； P <0.05; p <0.05; n = 3）。抗逆转录病毒没有影响24小时的细胞内和细胞外GH水平（方差分析； P> = 0.05; n = 3）。我们证明了抗逆转录病毒与细胞内钙的增加之间的关联，对体纤维细胞的生存能力和GH合成而没有影响。总体而言，这些结果表明，抗逆转录病毒可能不会直接影响受HIV感染的患者的GH轴。
Growth Hormone (GH) deficiency is frequent in HIV-infected patients treated with antiretroviral therapy. We treated GH3 cells with antiretrovirals (nevirapine, ritonavir or abacavir sulfate; 100 pM-1 mM range), after transfection with human growth hormone releasing hormone (GHRH) receptor cDNA. Cells viability, intracellular cAMP, phosphorylation of CREB and calcium increase, GH production and secretion were evaluated both in basal condition and after GHRH, using MTT, bioluminescence resonance energy transfer, western blotting and ELISA.Antiretroviral treatment did not affect GHRH 50% effective dose (EC50) calculated for 30-min intracellular cAMP increase (Mann-Whitney's U test; p >= 0.05; n = 4) nor 15-min CREB phosphorylation. The kinetics of GHRH-mediated, rapid intracellular calcium increase was perturbed by pre-incubation with drugs, while GHRH failed to induce the ion increase in ritonavir pre-treated cells (ANOVA; p < 0.05; n = 3). Antiretrovirals did not impact 24-h intracellular and extracellular GH levels (ANOVA; p >= 0.05; n = 3).We demonstrated the association between antiretrovirals and intracellular calcium increase, without consequences on somatotrope cells viability and GH synthesis. Overall, these results suggest that antiretrovirals may not directly impact on GH axis in HIV-infected patients.