Research On The In Vitro Blood-Brain Barrier Model (BBBM)as Neurotoxicity Screening Platform

Background:More and more chemicals coming into human living environment, chemical neurotoxicity hazard has been a serious public health-related problem. One effective way for chemical administration and chemical neuroxicity hazare control and prevention is to detect neurotoxic chemicals ahead of their hazard. Intrinsic complex of central nervous system(CNS), however, makes difficult for the distinguishment of neurotoxic chemical, which has been a big challenge for chemical neurotoxicity research and the control and prevention of chemical neurotoxic hazard. Recently, more attention was paid to the blood-brain barrier(BBB). BBB prevents exogenous chemicals from passing into CNS, which could safeguard CNS from exposing to excessive exogenous chemicals. On the other hand, if BBB can be damaged by certain exogenous chemicals, the barrier function will lose, and amounts of chemicals may easily invade into CNS, brain damage would be unavoidable. So, it’s rational to detect the neurotoxic potential of one exogenous chemical according to its capacity to damage BBB permeability. Though BBB characteristics are completely kept in in vivo animal test, from which results obtained are more credible, in vivo test is time-consumed, complicatedly performed, and not suit for highthrough toxicity screening. By comparison, in vitro models are easy prepared, efficient in toxicity screening. Presently, one technological trend aimed chemical neurotoxicity screening has been coming, that is, construction of BBB-based chemicals neurotoxicity screening platform by means of in vitro cell culture.Aim:To construct in vitro chemical neurotoxicity screening platform(CNSP) consisting of models including BBB permeability toxicity test model and astrocyte function test model and corresponding screening programme. And to detect neurotoxicity potential of15chemicals including Lead acetate, Lead nitrate, Aluminum sulfate, Magnesium chloride, Zinc Sulfate, Manganese Sulfate, Zinc acetate, Cadmium chloride, Magnesium sulphate, Stannous chloride, Sodium fluoride, Copper sulphate, Cadmium sulfate, Colchicine, and isopropyl alcohol utilizing CNSP to provide toxicity data.Methods:Cell culture(brain capillary endothelial cells(BCECs) and astrocytes(As) culture) was used to construct in vitro chemical neurotoxicity screening platform(CNSP), which consists of2-staged models including BBB permeability toxicity test model and astrocyte function test model according to study design. In vitro CNSP was used to detect neurotoxicity potential of15chemicals. Moreover,15chemicals were be grouped according screening results.Results:In vitro CNSP consisting of models including BBB permeability toxicity test model and astrocyte function test model(test parameters including glutamine synthetase(GS), glial cell line-derived neurotrophic factor(GDNF), and glucose transporter type1(GLUT1))was built up, and matching screening programme was formed meanwhile. Under testing circumstances of the platform, Lead acetate, Lead nitrate, Aluminum sulfate, Manganese Sulfate, Zinc acetate, Cadmium chloride, and Colchicine were detected BBB permeability toxicity-positive. Lead acetate, Lead nitrate, Aluminum sulfate, Manganese Sulfate, Cadmium chloride, and Colchicine could reduce GS in As to certain extent respectively. Lead acetate, Lead nitrate, Aluminum sulfate, Manganese Sulfate, Cadmium chloride, Colchicine and isopropyl alcohol could significantly affect on GDNF of As. And Lead nitrate, Aluminum sulfate, Manganese Sulfate, Zinc acetate, Cadmium chloride, Colchicine and isopropyl alcohol were detected effective on GLUT1expression in As.Conclusion:In vitro CNSP consisting of2-staged models including BBB permeability toxicity test model and astrocyte function test model and hierarchical screening programme is easily prepared, time-saved in use, and may effectively detect neurotoxic chemicals.15chemicals were tested through the screening platform, the results of which are useful toxicity data. Moreover,15chemicals were grouped according to their neurotoxicity potential detected, which is good for mastering neurotoxicity characteristics of chemicals, and taking effective measures to administer neurotoxic chemicals and control and prevent their toxicity hazard.