Effect Of Jiedu Yizhi Recipe On Neuroprotection And Aβ Peptide Aggregation Of Transgenic Nematodes Of AD

Objective: To observe the effect of xantholysaccharide on the neuroprotection of nematodes and the aggregation of A beta peptide in the clinical treatment of AD,the effects of xantholaccharide,an extract of xanthoxylophane,on the model of Alzheimer’s disease nematodes,were studied.Methods: Polysaccharides from Coptis chinensis were extracted from the freeze-dried powder of Jiedu Yizhi Formula prepared by our research group.Polysaccharides from Coptis chinensis were extracted by ultrasonic extraction,boiling extraction and cold soaking extraction.The optimum extraction conditions were investigated to obtain the extracts of Coptis chinensis polysaccharides with high purity.The model of genetically modified nematode of Alzheimer’s disease was prepared.Coptis chinensis was added to the experimental model.Polysaccharide extracts,including control group(containing Coptis chinensis polysaccharide 0 mg/L),low dose group(containing Coptis chinensis polysaccharide25 mg/L),medium dose group(containing Coptis chinensis polysaccharide 50 mg/L),high dose group(containing Coptis chinensis chinensis polysaccharide 100 mg/L),were used to investigate the effects of different concentration of drugs on nematode longevity,neuroprotection and aggregation of A beta peptide.Results:1.The yield of Polysaccharides from Coptis chinensis was 39.4% by ultrasonic extraction,42.7% by boiling extraction,22.8% by cold soaking extraction.2.The effects of Coptis chinensis polysaccharide on nematode longevity were studied in dose group(containing Coptis chinensis polysaccharide 25 mg/L),middle dose group(containing Coptis chinensis chinensis polysaccharide 50 mg/L),high dose group(containing Coptis chinensis chinensis polysaccharide 100 mg/L).The average survival rate of control group was 22.2-28.6%,that of 25 mg/L group was 63.0-72.3%,that of 50 mg/L group was48.9-62.5%,and that of 100 mg/L group was 59.3-72.03% and 5 days respectively.Theaverage survival rate of the control group was 5.6-10.2%,that of the 25 mg/L group was13.0-29.8%,that of the 50 mg/L group was 13.3-18.8%,and that of the 100 mg/L group was26.0-28.8%,the longevity level of nematodes in the 25 mg/L experimental group was similar to that in the control group.3.Paralysis rate of nematode model was tested under the same experimental grouping and concentration.From 36 h to 36 h,paralysis rate of nematode in each experimental group was higher than 90%.From 36-60 h,paralysis rate of nematode increased significantly.At 60 h,paralysis rate of nematode in 25 mg/L group was 50.0%,paralysis rate of nematode in 50mg/L group was 96.7%,paralysis rate of nematode in 100 mg/L group was 65.9%,paralysis rate of nematode in 60-92 H.The paralysis rate of tested nematodes decreased more obviously.At 92 hours,the nematodes in each experimental group were basically completely paralysed.Of them,6.7% of the nematodes in 100 mg/L experimental group did not occur paralysis.4.The experimental results of the effect of different concentrations of Coptis chinensis polysaccharides on the aggregation of Abeta peptide in nematode model showed that the aggregation of Abeta peptide in control group was very close,scattered holes could be seen in25 mg/L experimental group,and large fluorescent staining holes could be seen in 50 mg/L experimental group,which was more and more obvious than that in low-dose group.The phenomenon of fluorescent staining holes in 100 mg/L experimental group was more obvious,and some of the head of nematode was almost the same.Visible fluorescence staining.The fluorescence intensity of lysate was determined by dyeing after nematode lysis.The results showed that the fluorescence intensity of control group was 0.74±0.08,that of 25 mg/L experimental group was 0.71±0.06,that of 50 mg/L experimental group was 0.58±0.07,and that of 100 mg/L experimental group was 0.49±0.05,the increase of drug concentration,and the fluorescence intensity decreases gradually.Conclusion:1.Boiling method can obtain higher yield.The optimum extraction conditions are as follows: the ratio of solid to liquid is 1:3,the extraction temperature is 100 C,the extractiontime is 20 minutes,and the extraction times are 5 times.2.The results showed that Coptis chinensis polysaccharides had significant effects on the longevity of nematode model.There was no significant difference in the effect of 25-100mg/L concentration on the longevity of nematode model.3.The neuroprotective effect of Coptis chinensis polysaccharide on nematode was obvious.The neuroprotective ability increased with the increase of drug concentration in the range of 25-100 mg/L.4.It was determined that Coptis chinensis polysaccharide(CPS)with 25-100 mg/L could significantly inhibit the aggregation of A-beta peptide in AD nematode model,and the inhibition was positively correlated with the increase of drug concentration in the experimental concentration range.