Evaluation of HIV protease and nucleoside reverse transcriptase inhibitors on proliferation, necrosis, apoptosis in intestinal epithelial cells and electrolyte and water transport and epithelial barrier function in mice.
Evaluation of HIV protease and nucleoside reverse transcriptase inhibitors on proliferation, necrosis, apoptosis in intestinal epithelial cells and electrolyte and water transport and epithelial barrier function in mice.
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DOI：
10.1186/1471-230x-10-90
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发表时间：
2010-08-11
影响因子：
2.4
通讯作者：
Lima AA
中科院分区：
文献类型：
--
作者：
Braga Neto MB;Aguiar CV;Maciel JG;Oliveira BM;Sevilleja JE;Oriá RB;Brito GA;Warren CA;Guerrant RL;Lima AA
蛋白酶抑制剂和逆转录酶抑制剂是治疗人类获得性免疫缺陷综合征（AIDS）的高效抗逆转录病毒疗法（HAART）的重要组成部分。这些药物的长期临床疗效和治疗依从性受到不良副作用如腹泻的限制。本研究的目的是探讨选定的抗逆转录病毒药物对肠道组织病理学和功能在体内和细胞增殖和死亡在体外的影响。选定的抗逆转录病毒药物口服7天以上，瑞士小鼠，如下：100毫克/公斤的奈非那韦（NFV），茚地那韦（IDV），去羟肌苷（DDI）或50毫克/公斤的齐多夫定（AZT）。通过乳果糖和甘露醇测定法测量肠通透性;在灌注肠段中的净水和电解质转运;以及在处理和对照小鼠中评估小肠形态和细胞凋亡。使用WST-1试剂评价体外细胞增殖，并通过流式细胞术分析评价凋亡和坏死。NFV、IDV、AZT和DDI使十二指肠和空肠绒毛长度显著缩短（p < 0.05）。IDV和AZT增加十二指肠的隐窝深度，AZT增加空肠的隐窝深度。NFV、AZT和DDI显著降低回肠隐窝深度。所有选定的抗逆转录病毒药物显着增加净水分泌和电解质分泌，除了DDI，它不改变水或氯分泌。此外，只有NFV显著增加甘露醇和乳果糖的吸收。NFV和IDV在体外24 h和48 h均引起细胞增殖显著降低。DDI和AZT不改变细胞增殖。与对照组相比，70 μ g/mL NFV 24 h后IEC-6细胞的凋亡率显著增加（对照组：4.7% vs NFV：22%），而IDV、AZT和DDI在凋亡方面未显示出任何显著变化。在空肠切片中，IDV和NFV显著增加TUNEL阳性细胞的数量。PI的，NFV和IDV，增加细胞凋亡在体内，水和电解质的分泌和肠通透性和降低绒毛长度和细胞增殖。NFV是在体外增加细胞凋亡的唯一测试药物。核苷类逆转录酶抑制剂AZT和DDI不影响细胞凋亡或增殖。这些发现可以部分解释与PI相关的肠道副作用。
Protease inhibitors (PI's) and reverse transcriptase drugs are important components of highly active antiretroviral therapy (HAART) for treating human acquired immunodeficiency syndrome (AIDS). Long-term clinical therapeutic efficacy and treatment compliance of these agents have been limited by undesirable side-effects, such as diarrhea. This study aims to investigate the effects of selected antiretroviral agents on intestinal histopathology and function in vivo and on cell proliferation and death in vitro. Selected antiretroviral drugs were given orally over 7 days, to Swiss mice, as follows: 100 mg/kg of nelfinavir (NFV), indinavir (IDV), didanosine (DDI) or 50 mg/kg of zidovudine (AZT). Intestinal permeability measured by lactulose and mannitol assays; net water and electrolyte transport, in perfused intestinal segments; and small intestinal morphology and cell apoptosis were assessed in treated and control mice. In vitro cell proliferation was evaluated using the WST-1 reagent and apoptosis and necrosis by flow cytometry analysis. NFV, IDV, AZT and DDI caused significant reductions in duodenal and in jejunal villus length (p < 0.05). IDV and AZT increased crypt depth in the duodenum and AZT increased crypt depth in the jejunum. NFV, AZT and DDI significantly decreased ileal crypt depth. All selected antiretroviral drugs significantly increased net water secretion and electrolyte secretion, except for DDI, which did not alter water or chloride secretion. Additionally, only NFV significantly increased mannitol and lactulose absorption. NFV and IDV caused a significant reduction in cell proliferation in vitro at both 24 h and 48 h. DDI and AZT did not alter cell proliferation. There was a significant increase in apoptosis rates in IEC-6 cells after 24 h with 70 ug/mL of NFV (control: 4.7% vs NFV: 22%) while IDV, AZT and DDI did not show any significant changes in apoptosis compared to the control group. In jejunal sections, IDV and NFV significantly increased the number of TUNEL positive cells. The PI's, NFV and IDV, increased cell apoptosis in vivo, water and electrolyte secretion and intestinal permeability and decreased villus length and cell proliferation. NFV was the only drug tested that increased cell apoptosis in vitro. The nucleoside reverse transcriptase inhibitors, AZT and DDI, did not affect cell apoptosis or proliferation. These findings may partly explain the intestinal side-effects associated with PI's.