Neuroprotective Effect Of Senkyunolide H And Preparation And Characterization Of Its Nanoparticle

Ligusticum chuanxiong Hort.is a dicotyledonous plant belongs to Umbelliflorae,Umbelliferous family,and Ligusticum genus,which was widely distributed in Sichuan,Yunnan and Guizhou.As a traditional Chinese medicine,the root of L.chuanxing can be used to treat a variety of diseases,such as stroke,angioneurotic headache and rheumatic arthralgia.It has the functions of neuroprotection,promoting blood circulation,boosting Qi and tonifying collaterals.Among them,the neuroprotective effect of L.chuanxiong has attracted more and more attention,while few studies on the neuroprotective components has been reported.Therefore,the secondary metabolites were extracted from the roots of L.chuanxiong,the components with neuroprotective effect were screened and the underlying mechanism was discussed in this study.(1)The separation of senkyunolide H,senkyunolide I and ferulic acid were extracted from the root of L.chuanxiong by ethanol extraction method,separation,silicone column chromatography and gel column chromatography.(2)To confirm the effect and underlying mechanism of senkyunolide H,the in vitro Parkinson model was developed using MPP+-induced PC 12 cells.We demonstrated that senkyunolide H pretreatment significantly attenuated MPP+-induced neurotoxicity and apoptosis in PC 12 cells through cell viability,morphology,apoptosis-related factor expression,and oxidative stress-related parameters.Overall,this investigation provides novel evidence that senkyunolide H exerts neuroprotective effects via the ROS-mediated MAPK pathway and represents a potential preventive or therapeutic agent for neuronal disorders.(3)The in vitro OGD/R model was established by performing hypoxia and re-oxygenation treatment on PC 12 cells,which was used for considering the effect of senkyunolide H.After analyzation with network pharmacology.This study validates the expression level of PI3K-AKT-CREB pathway protein in cAMP downstream pathway,and finally we confirmed that senkyunolide H is activated by PI3K-AKT-CREB pathway and exerts neuroprotective effects.(4)In order to determine the improvement of senkyunolide H penetrating blood-brain barrier on damaged neurons,senkyunolide H-loaded liposomes were established.The encapsulation efficiency,size and Zeta potential,and morphological structure was observed,and the results indicated that the senkyunolide H-loaded liposomes showed small particle size and uniform distribution,with an average particle size of 61.13 nm and Zeta potential of-33.3 mV.The blank liposomes and SNH liposomes were spherical vesicles,and the encapsulation efficiency and drug loading were respectively 78%and 2.2%.