Cytotoxity of HIV-1 gp120 protein on human blood-retinal barrier ceils

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Author:
LIN Hao-tian(State Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China)
ZHANG Zhen-ping(State Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China)
YU Qiu-rong(State Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China)
YAN Pi-song(State Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China)
WANG Qi-lin(State Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China)
BAI Ling(State Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China)
Journal Title:
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
Issue:
Volume 15, Issue 01, 2009
DOI:
10.3760/cma.j.issn.1674-1927.2009.01.005
Key Word:
HIV envelope protein gp120;Blood- retinal barrier;Cytotoxicity tests;Apoptosis;Acquired immunodeficiency syndrome

Abstract: Objective To study the mechanism of human blood-retina barrier (BRB) destroyed by HIV- 1 gp120 protein. Methods Human blood-retina barrier cells (HBRBCs) including human retina capillary endothelial cells (HRCECs), human retina capillary perieytes (HRCPCs), human retinal pigment epithelium (HRPE) were primarily cultured. Culture media were regarded as eontrol. MTT method was used to observe the inhibition effect of HIV-1 gp120 protein on eell viability at 7 different concentrations (0.01 to 0.15 mg/L) for 24 h, and at a fixed concentration(0.08 mg/L) for different times (4-72 h). After 0.08, 0.1, 0.12 and 0.15 mg/L HIV- 1 gp120 protein were applied in those cells for 24 h, cell apoptotie rates and membrane potential of mitochondria (△ψm) were measured by flow eytometry. Activation of Cleaved caspase-9 was detected by Western blot. Change of cell mierostructure with 0.08mg/L HIV-1 gp120 protein before and after 24 h was detected by transmission electron microscopy (TEM). Results Concentration of HIV-1 gp120 protein less than 0.08 mg/L did not influence cell viability at 24 h. But at the concentration of more than 0.08 mg/L, HIV-1 gp120 protein could obviously inhibit HBRBCs proliferation with a concentration-dependent manner(HRCECs: r=-0.763, P<0.01 ; HRCPCs: r=-0.804, P<0.01 ; HRPE: r=-0.698, P<0.01). HIV-1 gp120 protein(0.08 mg/L) significantly inhibited cells proliferation at 12 h, and this inhibition effect was more stronger at 24,48,72 h with a time-dependent increase (HRCECs: r=-0.833, P<0.01 ; HRCPCs: r=-0.784, P<0.01 ; HRPE: r=-0.701, P<0.01). The relative growth rates were HRCECs: 84%, 70%, 41%, 22% ; HRCPCs: 80%, 69%, 38%, 18% ; HRPE: 86%, 73%, 45%, 26% respectively. Compared with control group, the ratio of apoptotic cells of HBRBCs and expression level of Cleaved caspase-9 protein increased but △ψm decreased at different concentrations with HIV-1 gp120 protein treatment for 24 h. The changes of these indexes were all concentration-dependent manner. Early apoptotic changes of HBRBCs such as mitochondrial swelling and lysosome increase were observed by TEM after treatment of 0.08 mg/L HIV- 1 gp120 protein for 24 h. Conclusion HIV-1 gp120 protein can inhibit proliferation and induce apoptosis of HBRBCs. The mechanism is probably associted with the structure and function destruction of mitochondiral.

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