Experimental Study Of Rat Hypothalamic Cells Transplantation To Therapy Opium Addiction

Object:Drug addiction has become an urgent question, and it's now seriously threatening humanity's health and stability of society. The mechanisms of drug addiction and its treatment methods have obtained some results at present. Drug addiction is an encephalopathy. It has been considered as a chronic disease like hypertension, asthma and non-insulin dependent diabetes. There are so many similar factors among them such as the heredity, personal liability, recurrent attacks and treatment response. But its social hazards are stronger. Therefore the opium addiction is one kind of diseases need medicine treat. The recent years research discovered, negative feedback adjustment of extrinsic opium medicine to endogenous β-endorphin (β-EP) system is possibly the important neurobiology mechanism of the opium addiction. With the technologydevelopment of cell graft, if we can transplant the active cells in vitro into opium addiction person in vivo, the implanted cells would like a pump continuously secrete P-EP. It can compensate self-secretion insufficiency of P-EP caused by opium addiction, and enable [3-EP reach the normal physiological level. This will become the ideal method to treat opium addiction. This research included: 1) Establish rat hypothalamic cells separation and foster methods, observe its cytomorphology characteristic in vitro and immunocytochemistry. The dynamic changes of secreting P-EP of cells in vitro were evaluated with ELISA. Find out self-adjust function of rat hypothalamic cells' (3-EP secretion, infer the opium addiction mechanism;2) Establish rat ileum morphine addiction model, observe the effect of hypothalamic cells culture to its contraction, and pinpoint the dependence function of rat hypothalamic cells secretion beta -EP anti- morphine;3) Establish rat model of morphine addiction, use celiac transplantation method of rat hypothalamic cells, discuss therapy function to opium addiction of rat hypothalamic cells celiac transplantation.Materials and methods:1) SD rat of newborn, dissection hypothalamic tissue. Make single cell fluid after digestion with pancreatin(0.25%), then inoculate to culture. The cells were identified with NSE immunocytochemical staining after 24 hours. Had cultured the hypothalamic cells for 9 days, in 1st, 3, 5, 7, 9 day take the cells under the inverted microscope observe the modality, changed the culture liquids every 3 days, collected culture liquids every day, then assaied [3-EP concentration;2) The cell separation and cultivate methods as part one. Puts the hypothalamic cells on the raise board, changed the culture liquids every 3 days, after 7 days obtain the serum. Takes the rat ileum, with Kreb ' s fluid contains morphine to establish the rat ileum specimen of morphine addiction model, use biological function record system record the change after grouping;3) The cell separation and raise methods as part one. Puts the hypothalamic cells on the cultivate board, changed the culture liquids every 3 days, after 7 days obtain the cell for use. Establish morphine addiction rat model, group stochastic. Observe weight and symptom changes of rat aftertransnlantatinnResult:I he hypothalamic culture contains neurocyte, neuroglia cell and blood vessel endothelial cell. The typical shape of cellular body under microscope has clear silhouette, round body and obvious halo around it at 7th day. The neuron cell body and protuberance also reach the peak length. The NSE positive cells of hypothalamus can be identified. Secrete of (3-EP was no significance changes with phases of experiment. Naloxone promote the rat ileum morphine addiction model contraction, the model of rat ileum appear slight contraction with hypothalamic culture, the model of rat ileum with no naloxone has not appeared contraction. The morphine addiction model rat's weight lightened after addiction, the weight rise after the model accomplish, especially in transplantation group. The scores of addiction is obviously improved after transplantation in morphine addiction rat model. Conclusions:Digest with pancreatin and cultivate the rat hypothalamic cells, we can get stable growing cells which have similar structure and immunocytochemistry with the cells in vivo. First day of cultivation, therat hypothalamic cells can secrete endorphin to a stable level, and maintain at this level. It can be concluded that the hypothalamic cells have good self-adjusted function on the secretion of (3-EP, also can steady secrete it with effective biology activeness. Through rat ileum morphine addiction model, we can confirm that the hypothalamic cells cultivated in vitro may provide the normal physiological dosage of f}-EP which needed in the process of opium addiction .The hypothalamic cells celiac transplantation of opium addiction rat's can continuing stably secrete endogenous opium substance, and substitute the extraneous opium substance when opium break off. It confirms the therapy feasibility of treat the opium addiction from the cell biological technology.