| Phillygenin(PHI)belongs to lignans,which are mainly derived from the leaves of Forsythia suspensa and Osmanthus fragrans.It has a variety of pharmacological activities,including its good antioxidant activity,anti-inflammatory activity,anti-hyperlipidemia and anti-tumor activity.This paper mainly studies the therapeutic effect and molecular mechanism of phillygenin on Parkinson’s disease by taking Parkinson’s disease(PD)model nematode as the research object and using the better antioxidant activity of phillygenin.The main research contents and results are as follows:(1)Firstly,the effect of phillygenin on PD was studied by using network pharmacology.By using the platform of Pharm Mapper database and Gene Gards database,156 intersection target proteins of phillygenin and Parkinson’s disease were obtained.The core target proteins such as IGF1,MAPK1,MAPK8,MAPK14 and SOD2 were obtained after further analysis of the MCODE plug-in in the String data analysis platform and Cytoscape 3.6.0software.The GO function and KEGG pathway of PHI and PD intersection target protein were further enriched and analyzed using DAVID database platform.The results predicted that the main molecular pathway of phillygenin regulating Parkinson’s disease might be completed through IIS in the life span pathway.(2)The effect of phillygenin on the physiological indexes of PD model nematode was studied.Put different concentrations of phillygenin(0,10,20,40 μg/m L),20 μg/m L and 40μg/m L phillygenin prolonged the life span of nematode,improved the movement ability of nematode,increased the accumulation of fat content in nematode,decreased the accumulation of synaptophysin in nematode,and enhanced the food perception behavior of BZ555 nematode and the expression of dopaminergic neurons.The results showed that phillygenin could improve the pathological characteristics of PD model nematode.(3)Study on the molecular mechanism of phillygenin on PD model nematode.Put different concentrations of phillygenin(0,10,20,40 μg/m L)after acting on TK22 nematode,μg/m L of phillygenin can prolong the life of the oxidative stress-sensitive TK22 nematode.After the occurrence of acute oxidative stress in NL5901 nematode induced by juglone,it was found that 20,40 μg/m L phillygenin can prolong the survival time of NL5901 nematode under acute oxidative stress.These results showed that phillygenin could reduce the level of oxidative stress in nematodes.The content of ROS and antioxidant enzyme in the nematode after the effect of phillygenin on the nematode was detected by using the kit for detecting the content of ROS and antioxidant enzyme,the results showed that phillygenin could reduce the content of ROS in nematode and promote the transcriptional expression of antioxidant enzymes.After the treatment of TJ356 and LD1 nematodes with 20 μg/m L of phillygenin,it was found that phillygenin could promote DAF-16 and SKN-1 from cytoplasm to nucleus.The results of q-PCR results showed that phillygenin increased the expression of daf-16,skn-1 and target genes gst-4,gcs-1 and sod-3,and decreased the expression of upstream key component daf-2.The binding of phillygenin with DAF-2,DAF-16,SKN-1,SOD-3,GCS-1 and GST-4 was analyzed by molecular docking software.Final result discoveryed that the binding energy was less than-5 k J/mol,indicating that phillygenin had good binding activity with these targets.To sum up,phillygenin can enhances the antioxidant stress ability of PD model nematode by regulating the gene expression of key regulatory factors in the insulin signal pathway,weaken the protein damage caused by ROS and thus weaken the pathological damage caused by Parkinson’s disease. |
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