The Influence Of AGEs And The Regulation Of Tonifying Kidney Recipe On The Learning And Memory In Ovariectomied Rats

Alzheimer's disease(AD) is a degeneration disorder of central neural system that significantly impairs the memory and cognitive functions of brain. Among the senior people, the pathologic character of AD is the formation of senile plaques in hemisphere, neurofibrillary tangles and the wide vacuolation of neurocyte. With the aging of the social population, AD has become one of the serious social problems that are highly thought of by the world. But the etiology of AD is still unknown, the advanced age and positive dementia family are considered as the protruded risk factor. Because of the aging and senile, the level of sex hormone gradually decrease, the effect of estrogen and AGEs related to the aging and senile in the pathogenesis of AD receive much attention. AGEs are compounds with characteristics of fluorescence that is produced by the non-enzymatic reaction of the free amino of the aldehyde groups or ketone groups of monosaccharides and proteins in body. The function of AGEs has been verified in many studies of senile diseases, and AGEs has been defined as the mark of senile biology in molecular level. It was reported that the content of AGEs in brain tissue of AD increased distinctly. The influence of AGEs on AD is probably due to the following reasons: ① The direct modification of protein can accelerate the aggregation of soluble Aβand antagonize the hydrolysis of protease, which results in the descent of normal updating metabolism and accumulation of abnormal protein and thus lose the normal structure and function. ② The pathway of receptor can enhance the oxidative stress and promote the degeneration of neurocyte. The generation of AGEs is complex, which is mainly related with the disorder of carbohydrate metabolism, oxidative stress and the produce of free radical. It has not been reported whether the descent of glucose tolerance and the enhancement of oxidative stress in postmenopausal woman because of the decrease of the level of estrogen can accelerate the generation of AGEs and the deposition of AGEs in brain and speed up the process of AD. According to traditional Chinese medicine, the kidney is the foundation of life, stores essence, dominates wisdom. The etiology of AD is deficiency of kidney, therefore AD has been treated with tonifying kidney.In order to study the pathologic mechanism of AD more, we adopt the model of ovariectomied rats and observe the content of AGEs in cerebrum cortex and hippocampi, and than, explore the relationship between the content of AGEs and the learning and memory. Moreover, we take AG—the specific inhibitor of AGEs, as the control to explore the pathologic significance of AGEs in the learning and memory in ovariectomied rats and the relationship between the AGEs and the learning and memory and the mechanism of tonifying kidney recipe(TKR) of preventing and curing AD. The experiment is divided into five parts.Part Ⅰ The preparation of AGEs and AGEs antibody in vitro Objective: To obtain high titer and specific polyclonal antibody of AGEs for immunochemical detection of AGEs.Method: BSA and glucose were incubated together in vitro to prepare AGEs-BSA, purified it through gel chromatography, immunize New Zealand white rabbit to obtain the anti-AGEs serum, and prepare the polyclonal antibody of AGEs through the sedimentation of saturate ammonium sulfate.Result:1. The evaluation of the purified AGEs-BSAThe purified AGEs-BSA possess special fluorescence spectrum and showed smear shape in SDS-PAGE.2. The specificity and titer of AGEs-BSA antibody AGEs-BSA antibody has strong specificity through the detection of ELISA and two-dimensional immunodiffusion, which has crossing reaction with AGEs-HAS but not with HSA. The titer of antibody by the detection of ELISA is 1: 2000.Conclusion: 1. AGEs-BSA can be successfully prepared through the incubation of glucose and BSA.2.The high titer of polyclonal antibody of AGEs can be successfully prepared by using AGEs-BSA to immunize New Zealand white rabbit and be used to detect AGEs by immun...