Vernochet Cécile, Azoulay Stéphane, Duval Danièle, Guedj Roger, Ailhaud Gérard, Dani Christian
Institute of Signalling, Development Biology and Cancer Research, Biochemistry Centre, UMR 6543 CNRS, Faculty of Science, Nice, France.
AIDS. 2003 Oct 17;17(15):2177-80. doi: 10.1097/01.aids.0000088160.01779.2b.
Lipodystrophy is a major side effect of HIV protease inhibitor (PI) antiretroviral therapy. It has been shown that protease inhibitors interfere in vitro with adipocyte differentiation. However, there is no evidence that PIs accumulate into preadipocytes and adipocytes and that intra-cellular accumulation is sufficient to alter differentiation. We assessed the effect of six different PIs on the differentiation of cells from four clonal lines. We also studied the capacity of ritonavir to accumulate both into drug-sensitive and drug-resistant cultured adipocytes.
Adipocyte differentiation of mouse 3T3-F442A, 3T3-L1 and Ob1771 cells as well as embryonic stem cells were investigated at pharmacological concentrations of indinavir, saquinavir, ritonavir, amprenavir, nelfinavir and lopinavir. We used a sensitive ELISA to determine intracellular concentration of ritonavir from 3T3-L1 and Ob1771 preadipocytes.
Nelfinavir and lopinavir inhibited adipocyte differentiation whereas amprenavir was ineffective. Indinavir, saquinavir and ritonavir inhibited differentiation of 3T3-L1 and 3T3-F442A cells but did not alter differentiation of either Ob1771 or embryonic stem cells. We showed that ritonavir accumulated in preadipocytes and fully differentiated 3T3-L1 adipocytes as a function of its extracellular concentration. Although Ob1771 cells were resistant and 3T3-L1 cells were sensitive to ritonavir, the drug accumulated to similar levels in both cases.
Protease inhibitors inhibit adipocyte differentiation depending on the cell model used. We showed for the first time that ritonavir accumulates into preadipocytes and adipocytes, suggesting a direct effect on intracellular targets. However, intracellular accumulation was clearly not sufficient as Ob1771 cells remained resistant to the inhibitory effect of ritonavir.
脂肪代谢障碍是HIV蛋白酶抑制剂（PI）抗逆转录病毒疗法的主要副作用。已有研究表明，蛋白酶抑制剂在体外会干扰脂肪细胞分化。然而，尚无证据表明蛋白酶抑制剂会在脂肪前体细胞和脂肪细胞中蓄积，且细胞内蓄积足以改变分化过程。我们评估了六种不同蛋白酶抑制剂对四个克隆系细胞分化的影响。我们还研究了利托那韦在药物敏感和耐药培养脂肪细胞中的蓄积能力。
在茚地那韦、沙奎那韦、利托那韦、安普那韦、奈非那韦和洛匹那韦的药理浓度下，研究小鼠3T3-F442A、3T3-L1和Ob1771细胞以及胚胎干细胞的脂肪细胞分化。我们使用灵敏的酶联免疫吸附测定法（ELISA）测定3T3-L1和Ob1771脂肪前体细胞中利托那韦的细胞内浓度。
奈非那韦和洛匹那韦抑制脂肪细胞分化，而安普那韦无效。茚地那韦、沙奎那韦和利托那韦抑制3T3-L1和3T3-F442A细胞的分化，但未改变Ob1771或胚胎干细胞的分化。我们发现利托那韦在脂肪前体细胞和完全分化的3T3-L1脂肪细胞中的蓄积与其细胞外浓度有关。尽管Ob1771细胞对利托那韦耐药，3T3-L1细胞对其敏感，但两种细胞中该药物的蓄积水平相似。
蛋白酶抑制剂对脂肪细胞分化的抑制作用取决于所使用的细胞模型。我们首次证明利托那韦会在脂肪前体细胞和脂肪细胞中蓄积，提示其对细胞内靶点有直接作用。然而，细胞内蓄积显然不足以使Ob1771细胞对利托那韦的抑制作用产生敏感性。
