| Background: Endogenous aldehyde(formaldehyde,ribose)has an important physiological function.Endogenous formaldehyde is participate in many kinds of cell process like the folic acid in the body circulation,DNA methylation,epigenetic modification,cell differentiation and gene expression regulation and so on.Endogenous ribose is participate in the synthesis of adenosie triphosphate(ATP)in the body,which is also an important part of the DNA and RNA.Metabolism disorder of aldehydes and excessive accumulation can reduce to of cognitive impairment.Experimental results show that endogenous formaldehyde increased with the aging process and associated with cognitive impairment.A significant rise of endogenous ribose in type 2 diabetes patient.Ribose and protein can produce AGEs quickly without enzymatic,which has cell cytotoxicity.However,whether low concentration formaldehyde treatment of nerve cells for a long time could lead to cell function damage,morphological change documents has not been reported.Whether insulin can rescue the nerve cell toxicity,cognitive impairment of AGEs produced by ribose has been reported.Objective: To study a long-term treatment of low concentration formaldehyde to nerve cells will reduce the change of morphological and function damage.Provide a new vision for studying the mechanism of Alzheimer’s disease and provide theoretical basis for early prevention,diagnosis and treatment.To explore whether insulin can rescue cell cytotoxicity and cognitive impairment of metabolism disorder of ribose.And to research the possible mechanism.Methods: Formaldehyde cause neurological chronic damage model is established by using a serial passage strategy and exposure to the pathological dose of formaldehyde.Using Holographic laser imaging system to observe the cell volume,area,thickness,roughness.Confocal microscope to observe the original number of dendrites and axons of neurons,the change of the length and complexity of dendrites and axons.Ribose reduce to cytotoxic damage model is established.CCK-8 to detect the cell vitality,crystal violet staining to detect cell number,holographic laser imaging system to detect cell morphological changes,Western blotting to detect expression changes of AGEs,insulin receptor,phosphorylation of insulin receptor and GRP78 protein.Cellular immune fluorescence detection which localization in cells and expression changes,ThT dyeing fiber entanglement detection in cells.High performance liquid chromatography(HPLC)to detect ribose content within the cell culture medium.Co-IP to detect whether there is interaction between the insulin receptor and AGEs in the cell.To establish the cognitive impairment animal model by intraperitoneal injection of ribose.Tensile test mice forelimbs.Morris water maze and Y maze test the ability of learning and memory in mice.Rotarod test the balance ability of mice.Western blotting and immunohistochemical testing organizations in glycosylation end products,insulin receptor,phosphorylation of insulin receptor,cathepsin-D and GRP78 protein expression changes.Biological experiment test in mice liver function,renal function,and insulin levels.Results: Long-term pathological FA exposure weakens the cells’ adhesive morphology and disrupts neurite morphology.Insulin could rescue the toxicity of ribose by active the insulin pathway,which is to eliminate AGEs. |
