The Effect And Mechanism Of Danggui-Shaoyao-San On APP/PS1 Alzheimer’S Disease Model Mice From The Perspective Of Lipid Metabolism

Objective The study was aim to observe the effects of extracting solution of Danggui Shaoyao San(DSS)on docosahexaenoic acid(DHA)metabolism,DHA metabolic signaling pathway neuroinflammation,Oxidative stress and the influence of cognitive function in the brain of APP/PS1 Alzheimer’s disease double transgenic mice,to discuss the mechanism of action of treat AD with DSS,to provide new methods for AD therapy.Methods First,the quality control research on DSS single herbs and the extracting solution of DSS.All the methods which measure the content of Paeoniflorin in White peony,ferulic acid in Angelica and Rhizome of Chuanxiong,23-acetyl alcohol B in Alisma are based on Chinese Pharmacopoeia 2015 edition,nand use the high-resolution mass spectrometry of flight time to qualitatively appraisal the seven kinds of active components of drugs in DSS water extract.Second,the influence of DSS on the mouse brain weight index、lipid levels 、learning and memory ability of the double transgenic AD model.Selected 6-month-old APP/PS1 transgenic mice(male)and screen them with Morris water maze,randomly divided the mice into model group(16),high,medium and low dose of DSS groups(11),positive medicine group(11)after elimination of unqualified animals,and A normal control group was selected for the transgenic male mice(16)with the same background.According to preliminary experimental results,the group of high,medium and low dose of DSS that gave mice daily gavage was 6.4 g/kg,3.2 g/kg and 1.6 g/kg respectively.The dosage was 3 mg/kg for the positive drug group(donepezil).The dosage was 10ml/kg for each drug group,and 10ml/kg of normal saline was given to the model group and control group.Gavage the mice once a day and Start behavioral testing after one month in a row.Weigh the mice the next day after the end of behavioral testing.Use 10% of chloral hydrate for intraperitoneal injection,take out blood when remove the eyeball and Centrifugally separated serum,then detect the TC,TG,LDL-C and HDL-C in serum,separate the whole brain and weigh it,calculate the brain weight index.Thirdly,the influence of DSS on DHA metabolism,nerve inflammation and oxidative stress in brain of APP/PS1 double transgenic AD model mice.Using LC-MS technology to test volume of unesterified DHA from the DHA enzyme metabolism,Using Western blotting to test Enzyme i PLA2,15-LOX released and metabolized from DHA through the cell membrane,and the rotein expression of enzyme c PLA2,COX-1,COX-2,5-LOX and 12-LOX,Which participate in the release and metabolism of inflammatory mediators.ELISA determined the inflammatory mediators PGE2,TXB2 and LTB4.Test kit was used to detect MDA,SOD,GSH and ROS in rats.Results 1.The quality control research on DSS single herbs and the extracting solution of DSS.The content of Paeoniflor in White peony,ferulic acid in Angelica and Rhizome of Chuanxiong,23-acetyl alcohol B in Alisma meets the standard of Chinese Pharmacopoeia 2015 edition,78 main components(in the negative ion mode,34 components were detected,and 44 components were detected in the positive ion mode)are detected from the extracting solution of DSS through the high-resolution mass spectrometry of flight time.For content determination of paeoniflorin in ester glycosides,paeoniflorin and benzoic acid,gallic acid,ferulic acid,rhizoma ligustici wallichii lactone I,rhizoma ligustici wallichii lactone A seven elements,content were 1.76,12.15,0.68,1.17,0.29,0.46,0.29 mg per gram extractum,and the highest content is paeoniflorin.2.The influence of DSS on the mouse brain weight index,lipid levels,learning and memory ability of the double transgenic AD model.The results of the AD model mouse brain weight index showed that: Compared with the control group,the proportion of brain in the model group decreased(P<0.01).The proportion of brain in high,middle,low DSS groups and positive drug control group was increased(P<0.01)The results of serum lipid level in AD model mice showed that: Compared with the control group,the TC,TG,and LDL-C levels in the model group all increased(P<0.01),HDL-C levels decreased(P<0.01);DSS high,middle,Low-dose group and positive drug group can significantly improve the lipid levels in AD model mice(P<0.05 or P<0.01).The behavioral results of AD model mice showed that:The result of Morris water maze positioning navigation showed that compared with the normal group,the latency period of the APP/PS1 model group with the same background and monthly age was significantly prolonged(p<0.05);Positive drug group(P<0.05)and high,middle and low DSS group(P<0.01)significantly shortened the latency of AD model mice.The result of Morris water maze space exploration showed that compared with the control group,the first time of the model group was significantly extended(P<0.01)and the number of crossing platforms was significantly reduced(P<0.01);the positive drug group and the high,middle,low DSS group could significantly shorten the time of the first crossing platform of the AD model mice(P<0.01)and increase the number of crossing platforms(P<0.05 or P<0.01).The result of Y maze showed that compared with the control group,the alternation percentage of the model group in the three arms decreased significantly(P<0.01).In the high,middle,low DSS group,the percentage of the mice in the model group was increased(P<0.01).The result of field experiment showed that compared with the control group,the time of the model group entering the central region decreased significantly(P<0.01).In the positive drug group and the high,middle,low DSS group,the time of the rat in the central area was increased(P<0.01).The result of conditioned fear experiment showed that compared with the control group,the lag time of the model group was significantly shorter(P < 0.01).The stagnation time of model mice(P<0.01)was increased in both the positive and DSS groups.3.The influence of DSS on DHA metabolism,nerve inflammation and oxidative stress in brain of APP/PS1 doubles transgenic AD model mice.Compared to the model group,there was a significant increase in the amount of DHA(P<0.05)in the brain of rats with high,middle,low DSS group and positive drug group.In addition to low DSS,positive drug group,high DSS group and middle DSS group can obviously increase the mice hippocampus and cortex in DHA have nerve protection by metabolic enzymes and antioxidant effect of NPD1—15-LOX and i PLA2 protein expression(P<0.05 or P< 0.01),lower involved in the release of inflammatory mediators,metabolic enzyme c PLA2,COX-1,COX-2,5-LOX and 12-LOX protein expression(P < 0.05 or P < 0.05),However,the expression of 15-LOX and i PLA2 in hippocampus was significantly increased in DSS low-dose group,and the expression of 5-LOX and 12-LOX in hippocampus and cortex(P<0.05 or P<0.01)and the expression of COX-1 in cortex(P<0.05).Compared with the model group,the positive drug group,the high,middle and low DSS group all can make the neuroinflammatory mediators PGE2,TXB2 and LTB4 in the hippocampal and cortex of the AD model decreased significantly(P<0.05 or P<0.01).The positive medicine group,the high,middle,and low DSS group can make the AD model mice hippocampus,cortex MDA and ROS reduced significantly(P < 0.05 or P < 0.01),positive medicine group,the high,middle,and low DSS group can increase the SOD activity of mice hippocampus and cortex(P < 0.05);The GSH content in the hippocampal and cortex of the mice was reduced(P<0.05 or P<0.01)(except in DSS),with the positive drug group and the high、middle and low DSS group.Conclusion The high,middle and low DSS group of extracting solution of Danggui Shaoyao San(DSS)all can improve the learning and memory disorders of the APP/PS1 double transgenic AD model mice,improve the blood lipid level,improve the AD model mice in the hippocampus and cortex and esterification of DHA content increase DHA by related metabolic enzymes to produce NPD1—15-LOX and i PLA2 protein expression,reduce the neurotransmitter PGE2,TXB2 and the formation of LTB4,cut with neurotransmitters and generate related c PLA2,12-LOX,5-LOX,COX-1 and the expression of COX-2 protein,and can also improve the oxidative stress state in the APP/PS1 Alzheimer’s disease double transgenic mice.