| Objective:In this experiment,Zuoguiyin,the typical prescription to invigorating kidney yin,is as the research object,and through combining TCM clinical efficacy with modern pharmacological and metabonomic research methods,we study to delay the aging of liver and kidney of rats and its mechanism of action,then providing more experimental basis for the theory of anti-aging of Zuoguiyin,but also providing scientific guidance for its clinical medication.Methods:In this experiment,the experimental rats were divided into the normal control group,the model group and the drug groups(the vitamin E group,the Liuweidihuangwan group,the Zuoguiyin high dose group,the Zuoguiyin low dose group),using the D-galactose to build subacute aging model of experimental rats,at the same time,the drug groups were given corresponding intervention drug for 8 weeks,the vitamin E group as western medicine positive control group,the Liuweidihuangwan group as positive control group of Chinese medicine,then studying on the Zuoguiyin high dose group and the Zuoguiyin low dose group.Experimental research methods are as follows:(1)Behavioral experiment(including Morris water maze test and platform test)was used to evaluate the rats model of aging,to investigate the learning and memory ability of rats;(2)Measure the liver function index and free radical metabolism index in liver and kidney of rats,then studying the effects of Zuoguiyin on liver and kidney function and free radical metabolism in aging model rats;(3)Pathological sections were used to observe the liver and kidney sections of the senescent model rats,and objective to study the influence of Zuoguiyin on the morphological structure and pathological changes of liver and kidney tissue in aging model rats;(4)Using immunohistochemistry on apoptosis related protein Bax,Bcl-2 and Caspase-3 were measured,then studying the effects of Zuoguiyin on pathological changes of liver and kidney,and apoptosis related protein expression level in aging rats;(5)By metabonomic method and LC-MS technique to analyze the urine of model rats,to compare of Zuoguiyin callback effect on endogenous biomarkers,then studying the effect of Zuoguiyin on aging rats urine metabonomics.Results:(1)In the behavioral evaluation of rats,Zuoguiyin can shorten the latency time of rats in water maze and step-down test,and increase the number of hidden platforms through water maze test,and reduce the number of platform test errors;(2)In biochemical tests,besides ALT,reduced the effects of Zuoguiyin on AST,ALP,CREA and BUN index level in rats serum,showed a trend of obvious correction;Zuoguiyin could increase the SOD and T一AOC activity in liver and kidney of rats,and reduce the content of MDA;(3)Observing the pathological changes of liver and kidney,Zuoguiyin could alleviate the pathological damage of liver and kidney cells in a certain extent,and can keep the integrity of the structure;(4)In immunohistochemical test,Zuoguiyin could enhance the expression of anti apoptosis protein Bcl-2,and reduce the expression of promote apoptosis protein Bax and Caspase-3;(5)In the rats urine metabolomics experiment,based on the urine metabolic profile of rats model,identified a total of 29 biomarkers and 9 related metabolic pathways,Zuoguiyin can make the urine metabolism profile in rats model returning to normal,have a callback function to 23 biomarkers and effect 7 related metabolic pathways.Conclusions:Zuoguiyin can improve the learning and memory ability of aging rats,improve liver and kidney function in aging model rats,increase of free radical in vivo aging rats metabolism and scavenging ability,protect liver and kidney cells from injury,exert anti-aging effect.The effect of delaying aging may regulate the expression of related apoptosis proteins in liver and kidney tissues,and regulate the abnormal metabolism of aging model rats. |
