Protective Effect And Mechanism Of Decoction Of Bidens Pilosa L.on Non-alcoholic Fatty Liver

OBJECTIVE:To investigate the preventive effect and possible mechanism of decoction of Bidens pilosa L.on non-alcoholic fatty liver induced by high fat and sugar in mice.METHODS:100 Balb/c mice were adaptively fed about 1 week,80 mice were randomly selected and fed with high-fat and high-sugar diet for 8 weeks to establish a non-alcoholic fatty liver（NAFLD）model,and the remaining 20 mice were fed with regular feed;Serological indicators and pathological tissues of NAFLD model were measured,and then the NAFLD models that met the selection requirements were randomly divided into model group,metformin(200mg·Kg^-1)treatment group,and decoction of Bidens pilosa L.(10g·Kg^-1)treatment group,metformin,and Bidens pilosa L.combined treatment group(100mg·Kg^-1+5g·Kg^-1),each treatment group was given an intragastric administration at 8 am every day,and the normal group and the model group were given an equal volume of normal saline,during the treatment period,except for the normal group,the other groups were still fed with high-fat and high-sugar feed;after 4 weeks of treatment,blood was taken from the eyeballs and the serum was quickly separated after 8 hours of fasting to detect serological indicators;mice were sacrificed,the liver tissue was taken and divided into three parts,the liver lobules from the same part of each mouse were subjected for oil red O staining experiments,the liver lobes from the same part were fixed for HE staining and immunohistochemical staining,the remaining liver tissue was stored at-80°C for related factors by Western blot experiments.RESULTS:（1）Results of serological indicators:Compared with the normal group,the levels of cholesterol（TC）,triglyceride（TG）,LDL-cholesterol（LDL-C）and blood glucose（FBG）in the model group were significantly increased（P<0.01）,HDL-cholesterol（HDL-C）was significantly reduced（P<0.01）;compared with the model group,the TC,TG,LDL-C and FBG of each treatment group were significantly reduced（P<0.05 or P<0.01）,HDL-C was sigificantly increased（P<0.05 or P<0.01）;compared with the metformin treatment group,the blood lipid level and blood glucose level of the combined medication group were significantly changed（P<0.05 or P<0.01）;compared with the treatment group of decoction of Bidens pilosa L.,the levels of blood lipids and blood glucose were significantly changed in the combination group（P<0.05 or P<0.01）;（2）Liver morphology:The liver of the normal group is soft and moist,with bright color and smooth surface,the liver of the model group is rough,tarnished,pale and yellowish,and brittle,the appearance of the liver in each treatment group was improved,and the liver in the combination treatment group was significantly improved;（3）HE staining:In the normal group,the liver tissue structure is clear,the cells are evenly distributed,and it has typical cell structure characteristics,the cell membrane and the liver lobules are intact,the tissue structure of the model group was disordered,and the structure of the hepatic lobules was damaged,the tissue structure of each treatment group was restored,and the effect of the combined drug group was better;（4）Oil red O staining results:Compared with the normal group,the fat distribution in the liver tissue of the model group was significantly increased（P<0.01）,compared with the model group,the fat distribution in each treatment group was significantly reduced（P<0.01）,compared with the metformin treatment group,the fat distribution in the combination group was significantly reduced（P<0.01）,compared with the treatment group of decoction of Bidens pilosa L.,the fat distribution of the combined medication group was significantly reduced（P<0.01）;（5）Immunohistochemical staining results:Compared with the normal group,the expression of GRP78 in the model group was significantly increased（P<0.01）,compared with the model group,the expression of GRP78 protein in each treatment group was significantly reduced（P<0.01）,compared with the metformin treatment group or the decoction of Bidens pilosa L.treatment group,the expression of GRP78in the combination group was significantly reduced（P<0.01）;（6）Western Blot results:Compared with the normal group,the protein expression levels of endoplasmic reticulum stress signaling pathway-related factors GRP78,p-PERK,e IF2α,ATF4,CHOP,IRE1α,and Cleaved-caspase12 in the model group were significantly increased（P<0.01）,and Bcl-2 expression was significantly reduced（P<0.01）;Compared with the model group,the expression of endoplasmic reticulum stress signaling pathway-related proteins in each treatment group was significantly reduced（P<0.01）,and Bcl-2 expression was significantly increased（P<0.01）;Compared with the monotherapy group,the expression of each protein in the combination group was more significant（P<0.01）.CONCLUSION:The decoction of Bidens pilosa L.has significant therapeutic effect on non-alcoholic fatty liver caused by high fat and sugar in mice,and metformin and decoction of Bidens pilosa L.combined treatment of NAFLD is more effective than monotherapy.The mechanism may be related to down-regulating the expression of endoplasmic reticulum stress-related factors,reducing the apoptosis of hepatocytes caused by endoplasmic reticulum stress,and down-regulating blood lipid and blood glucose levels.It provides experimental basis and theoretical basis for clinical application of Bidens picata L.,and lays a foundation for combined use of Bidens picata L.and metformin.