The Effects Of Endoplasmic Reticulum Stress Induced By Homocysteine Thiolactone On Vascular Dementia

Objective:This research is designed to test whether homocysteine thiolactone elevated cell apoptosis and inflammatory in vascular dementia,caused by activating caspase-12,one pathway of endoplasmic reticulum stress.Meanwhile,we can measured the level of caspase-12 and other protein related to endoplasmic reticulum stress(caspase-3 and TNF-α)in clinical.Methods:①Basic medical experiment:we cultured SHSY5Y cells then treaed with different concentrations of homocysteine thiolactone(0,50,100,200 and 400 μmol/L).3h later,we observe the change of cell morphology through MTT assay.Futher,we make a vascular dementia model by CoCl2(the concentration of the original liquid is O.lmol/L 30μl+ 15ml DMEM),which treat SHSY5Y cells 24h.well HTL treated cells in the same way.So,we can get the most appropriate of concentration and time.Last,to detect the relationship between inflammatory response and caspase-12 pathway by flow cytometry and immunohistochemistry.② Clinical experiments:to collect vascular dementia and control group patients from first affiliated hospital of Beihua University during November 2015 to November 2017,two groups of influence factor like age,gender are no statistical difference.Meanwhile,the VD patients were made a mental state examination of Hachinski ischemia scale,MMSE and MoCA to be divided in several groups.Then,to collect the serum of two groups of patients,using ELISA to test caspase-12,caspase-3 and TNF-α,making a statistical analysis by SPSS17.0.Results:the different concentrations of HTL can inhibit the growth of SHSY5Y cells more or less after 3h,and 200μmol/L and 400μmol/L have a better effect.Also,with the increase of concentration of HTL,caspase-12 and caspase-3 increased,TNF-a showed a growing tendency as well,suggested that there was a relationship between HTL and three proteins of endoplasmic reticulum stress.In clinical,the serum of caspase-12,caspase-3 and TNF-α was all improved in the group of VD,considering there was some connection between patients’ cognitive dysfunction and three kinds of proteins.In conclusion,HTL may contribute to the level of cognitive dysfunction of VD patients by inducing caspase-12 pathway of endoplasmic reticulum stress,which is may involved TNF-α.Conclusion:1.The low concentration of HTL could inhibit growth of SHSY5Y and hypoxia cell model(CoCl2+SHSY5Y cells)cells,which is depend on concentration;2.HTL could accelerate cell poptosis by caspase-12 pathways of endoplasmic reticulum stress,which involved TNF-α;3.The VD group’s serum of caspse-12,caspase-3 and TNF-α level is higher than the control group;4.Hcy can expedite the level of VD patients’ cognitive dysfunction,which is related to caspase-12,caspase-3 and TNF-α;...