Damage And Mechanisms Of Neuron Induced By Tenofovir Ester

Objective:Tenofovir ester solution apply to SH-SY5 Y cells,neurons and mouse neurons to observe the effects on cell survival,apoptosis and related indicatorsat different concentration of tenofovir ester solutionculturedin vitro.A preliminary discuss of Tenofovir ester of nerve damage,mechanism of action and tenofovir ester long-term relationship with HIV-associated dementia research in different types of cells as model.Methods:MTT colorime is used to detect tenofovir estereffect of cell proliferation.Enzyme standard method detect the activity of LDH in cell cultures by tenofovir ester.Tenofovirester change characteristics in form of apoptosis is observed by Hoechst-33342 staining.In situterminal transferase labeling method(TUNEL)and Annexin V/PI labeling staining flow cytometry instrument applyto test cell apoptosis by tenofovir ester.Malondialdehyde(MDA)test kit measure intracellular malondialdehyde(MDA)content.DCFHDA staining method detect intracellular reactive oxygen species(ROS)level.TUNEL method detect tenofovir esterinfluence on apoptosis of brain slice.Caspase-3 kits apply to detect the intracellular activity of Caspase-3 by tenofovir.Results:MTT method and LDH method turn out that tenofovir ester inhibit the growth of SH-SY5 Y cells and neurons,themore increase of concentration of tenofovir ester,the more obvious inhibitor(P<0.05),suggest that tenofovir ester oxidative damage cells.Tenofovir ester changeSH-SY5 Y cells and neuron cells' morphology,the features of apoptosis such as specific cash for chromatin condensation,the nucleus concentration and marginalize,apoptotic bodies.TUNEL and Annexin V/PI labeling staining flow cytometry instrument detection showthat 5 umol/L,10 umol/L,20 umol/L,40 umol/L tenofovirester treat-ed after 72 h cells apoptosis rate significant higher comparewith the control group(P<0.01).Cell oxidation-antioxidant substances or related enzyme are changed,ROS and MDA content are significantincreased,Caspase-3 activation is also significantincreased(P < 0.01).A large number of Small mouse nerve cells are apoptosis after 72 hby 5 umol/L,10 umol/L,20 umol/L,40 umol/L tenofovirester,and they are depend on the concentration(P < 0.01).Conclusion:(1)SH-SY5 Y and nervecells' survival rateandlevels of LDH are reducedby tenofovir ester.So Tenofovir ester has obvious toxicityin nerve.(2)Tenofovir esterdamage neurological embody induce of cellapoptosis and depend on of tenofovir's concentration.(3)Tenofovir ester induce of neuronal apoptosis mechanism is mainly generate a lot of ROSand MDA,Caspase-3 activation is also increased,depend on of tenofovir's concentration.