Institute of Biomedicine and Clinical Research Unit-University Hospital, Federal University of Ceará, Fortaleza, Brazil.
BMC Gastroenterol. 2010 Aug 11;10:90. doi: 10.1186/1471-230X-10-90.
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.
蛋白酶抑制剂（PI）和逆转录酶药物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治疗（HAART）的重要组成部分。这些药物的长期临床治疗效果和治疗依从性受到不良副作用的限制，例如腹泻。本研究旨在研究选定的抗逆转录病毒药物对体内肠道组织病理学和功能的影响，以及对细胞增殖和死亡的体外影响。
选择抗逆转录病毒药物，通过口服方式给予瑞士小鼠，以下是具体的剂量：奈非那韦（NFV）100mg/kg、茚地那韦（IDV）、去羟肌苷（DDI）或齐多夫定（AZT）50mg/kg。通过乳果糖和甘露醇测定法测量肠道通透性；通过灌流肠段测量净水和电解质转运；在处理和对照小鼠中评估小肠形态和细胞凋亡。使用 WST-1 试剂评估细胞增殖，通过流式细胞术分析评估细胞凋亡和坏死。
NFV、IDV、AZT 和 DDI 均导致十二指肠和空肠绒毛长度显著降低（p < 0.05）。IDV 和 AZT 增加了十二指肠隐窝深度，而 AZT 增加了空肠隐窝深度。NFV、AZT 和 DDI 显著降低了回肠隐窝深度。所有选定的抗逆转录病毒药物均显著增加了净水分泌和电解质分泌，但 DDI 没有改变水或氯化物分泌。此外，只有 NFV 显著增加了甘露醇和乳果糖的吸收。NFV 和 IDV 在 24 小时和 48 小时均显著降低了体外细胞增殖。DDI 和 AZT 未改变细胞增殖。在 70μg/mL NFV 下，IEC-6 细胞的凋亡率在 24 小时后显著增加（对照：4.7% vs NFV：22%），而 IDV、AZT 和 DDI 与对照组相比，凋亡没有明显变化。在空肠切片中，IDV 和 NFV 显著增加了 TUNEL 阳性细胞的数量。
PI 类药物奈非那韦和茚地那韦增加了体内细胞凋亡、水和电解质分泌以及肠道通透性，降低了绒毛长度和细胞增殖。NFV 是唯一一种在体外增加细胞凋亡的药物。核苷逆转录酶抑制剂 AZT 和 DDI 均不影响细胞凋亡或增殖。这些发现可能部分解释了与 PI 相关的肠道副作用。
