Lopinavir inhibits insulin signaling by promoting protein tyrosine phosphatase 1B expression

Kitazawa,Takatoshi; Yoshino,Yusuke; Suzuki,Satoshi; Koga,Ichiro; Ota,Yasuo

doi:10.3892/etm.2014.1826

Lopinavir inhibits insulin signaling by promoting protein tyrosine phosphatase 1B expression

Authors:
- Takatoshi Kitazawa
- Yusuke Yoshino
- Satoshi Suzuki
- Ichiro Koga
- Yasuo Ota
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Affiliations: Department of Medicine, Teikyo University School of Medicine, Tokyo 173‑8605, Japan, Department of Pulmonary Medicine,Graduate School of Medicine, The University of Tokyo, Tokyo 113-8655, Japan
Published online on: July 4, 2014 https://doi.org/10.3892/etm.2014.1826
Pages: 851-855

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Abstract

Treatment with antiretroviral therapy, including protease inhibitors (PIs), may result in metabolic side‑effects, for example insulin resistance. The aim of the present study was to investigate the mechanism of the dysregulation of insulin signaling by two PIs, lopinavir and darunavir, by analyzing changes in the expression or activity of proteins associated with insulin signaling. 3T3-L1 preadipocytes were pretreated with lopinavir or darunavir for 48 h and then stimulated with insulin for 30 min. The cell lysates were subjected to western blotting with anti‑phospho‑insulin receptor substrate (IRS) 1, anti‑IRS1, anti‑suppressor of cytokine signaling (SOCS) 1, anti‑SOCS3 and anti‑protein tyrosine phosphatase (PTP) 1B antibodies and to immunoprecipitation with anti‑IRS1 antibody. Translocation of glucose transporter 4 (GLUT4) following treatment with lopinavir or darunavir was observed using immunofluorescence. While GLUT4 was recruited to the cellular membrane in control adipocytes following insulin stimulation, it was diffusely distributed in the cytosol in lopinavir‑treated adipocytes. In darunavir‑treated adipocytes, GLUT4 was mainly recruited to the cellular membrane, but some GLUT4 remained in the cytosol. After insulin stimulation, IRS1 was tyrosine‑phosphorylated to a greater extent in control adipocytes compared with darunavir‑treated adipocytes. Tyrosine phosphorylation of IRS1 was inhibited in lopinavir‑treated adipocytes. The expression of PTP1B was upregulated in adipocytes pretreated with the PIs, particularly lopinavir, compared with those pretreated with a vehicle control. The degree of regulation in insulin signaling differs between lopinavir and darunavir. One mechanism by which lopinavir regulates insulin signaling is by the promotion of PTP1B expression.

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1	May MT, Sterne JA, Costagliola D, et al; Antiretroviral Therapy (ART) Cohort Collaboration. HIV treatment response and prognosis in Europe and North America in the first decade of highly active antiretroviral therapy: a collaborative analysis. Lancet. 368:451–458. 2006. View Article : Google Scholar : PubMed/NCBI
2	Honda M and Oka S: Current therapy for human immunodeficiency virus infection and acquired immunodeficiency syndrome. Int J Hematol. 84:18–22. 2006. View Article : Google Scholar : PubMed/NCBI
3	Murata H, Hruz PW and Mueckler M: The mechanism of insulin resistance caused by HIV protease inhibitor therapy. J Biol Chem. 275:20251–20254. 2000. View Article : Google Scholar : PubMed/NCBI
4	Nolte LA, Yarasheski KE, Kawanaka K, Fisher J, Le N and Holloszy JO: The HIV protease inhibitor indinavir decreases insulin- and contraction-stimulated glucose transport in skeletal muscle. Diabetes. 50:1397–1401. 2001. View Article : Google Scholar
5	Djedaini M, Peraldi P, Drici MD, et al: Lopinavir co-induces insulin resistance and ER stress in human adipocytes. Biochem Biophys Res Commun. 386:96–100. 2009. View Article : Google Scholar : PubMed/NCBI
6	Wellen KE and Hotamisligil GS: Inflammation, stress, and diabetes. J Clin Invest. 115:1111–1119. 2005. View Article : Google Scholar : PubMed/NCBI
7	Weisberg SP, McCann D, Desai M, Rosenbaum M, Leibel RL and Ferrante AW Jr: Obesity is associated with macrophage accumulation in adipose tissue. J Clin Invest. 112:1796–1808. 2003. View Article : Google Scholar : PubMed/NCBI
8	Dalpke A, Heeg K, Bartz H and Baetz A: Regulation of innate immunity by suppressor of cytokine signaling (SOCS) proteins. Immunobiology. 213:225–235. 2008. View Article : Google Scholar : PubMed/NCBI
9	Ueki K, Kondo T and Kahn CR: Suppressor of cytokine signaling 1 (SOCS-1) and SOCS-3 cause insulin resistance through inhibition of tyrosine phosphorylation of insulin receptor substrate proteins by discrete mechanisms. Mol Cell Biol. 24:5434–5446. 2004. View Article : Google Scholar
10	Asante-Appiah E and Kennedy BP: Protein tyrosine phosphatases: the quest for negative regulators of insulin action. Am J Physiol Endocrinol Metab. 284:E663–E670. 2003.PubMed/NCBI
11	Tordjman KM, Leingang KA, James DE and Mueckler MM: Differential regulation of two distinct glucose transporter species expressed in 3T3-L1 adipocytes: effect of chronic insulin and tolbutamide treatment. Proc Natl Acad Sci USA. 86:7761–7765. 1989. View Article : Google Scholar
12	Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227:680–685. 1970. View Article : Google Scholar : PubMed/NCBI
13	Eron JJ, Feinberg J, Kessler HA, et al: Once-daily versus twice-daily lopinavir/ritonavir in antiretroviral-naive HIV-positive patients: a 48-week randomized clinical trial. J Infect Dis. 189:265–272. 2004. View Article : Google Scholar : PubMed/NCBI
14	Curran A, Gutirerrez M, Deig E, et al: Efficacy, safety and pharmacokinetics of 900/100 mg of darunavir/ritonavir once daily in treatment-experienced patients. J Antimicrob Chemother. 65:2195–2203. 2010. View Article : Google Scholar : PubMed/NCBI
15	Patick AK and Potts KE: Protease inhibitors as antiviral agents. Clin Microbiol Rev. 11:614–627. 1998.PubMed/NCBI
16	Bogachus LD and Turcotte LP: HIV protease inhibitors induce metabolic dysfunction in part via increased JNK1/2 pro-inflammatory signaling in L6 cells. Antiviral Res. 92:415–423. 2011. View Article : Google Scholar : PubMed/NCBI
17	Sun XJ and Liu F: Phosphorylation of IRS proteins Yin-Yang regulation of insulin signaling. Vitam Horm. 80:351–387. 2009.PubMed/NCBI
18	Ismail WI, King JA, Anwar K and Pillay TS: Indinavir and nelfinavir inhibit proximal insulin receptor signaling and salicylate abrogates inhibition: potential role of the NFkappa B pathway. J Cell Biochem. 114:1729–1737. 2013. View Article : Google Scholar : PubMed/NCBI
19	Goldstein BJ, Bittner-Kowalczyk A, White MF and Harbeck M: Tyrosine dephosphorylation and deactivation of insulin receptor substrate-1 by protein-tyrosine phosphatase 1B. Possible facilitation by the formation of a ternary complex with the Grb2 adaptor protein. J Biol Chem. 275:4283–4289. 2000. View Article : Google Scholar
20	Ben-Romano R, Rudich A, Tirosh A, et al: Nelfinavir-induced insulin resistance is associated with impaired plasma membrane recruitment of the PI 3-kinase effectors Akt/PKB and PKC-zeta. Diabetologia. 47:1107–1117. 2004. View Article : Google Scholar : PubMed/NCBI
21	Schütt M, Zhou J, Meier M and Klein HH: Long-term effects of HIV-1 protease inhibitors on insulin secretion and insulin signaling in INS-1 beta cells. J Endocrinol. 183:445–454. 2004.PubMed/NCBI
22	Noor MA, Flint OP, Maa JF and Parker RA: Effects of atazanavir/ritonavir and lopinavir/ritonavir on glucose uptake and insulin sensitivity: demonstrable differences in vitro and clinically. AIDS. 20:1813–1821. 2006. View Article : Google Scholar : PubMed/NCBI
23	Tomaka F, Lefebvre E, Sekar V, et al: Effects of ritonavir-boosted darunavir vs. ritonavir-boosted atazanavir on lipid and glucose parameters in HIV-negative, healthy volunteers. HIV Med. 10:318–327. 2009. View Article : Google Scholar : PubMed/NCBI
24	Björnholm M, Kawano Y, Lehtihet M and Zierath JR: Insulin receptor substrate-1 phosphorylation and phosphatidylinositol 3-kinase activity in skeletal muscle from NIDDM subjects after in vivo insulin stimulation. Diabetes. 46:524–527. 1997.PubMed/NCBI