Study On The Efficacy And Mechanism Of Polygonatum Kingianum On Relieving Nonalcoholic Fatty Liver Based On Mitochondrial Function Regulation

BackgroundMitochondrial dysfunction is an important pathogenesis of non-alcoholic fatty liver disease(NAFLD).The development of mitochondrial modulators/nutrients from natural products to treat mitochondrial dysfunction is an effective strategy for the prevention and treatment of NAFLD.Polygonatum kingianum(PK)is an important dual-use resource for medicine and food.The previous research of this group showed that the active ingredient group(total polysaccharide,total saponin)can alleviate type2 diabetes,and its water extraction can also regulate abnormal lipid metabolism.However,it is still unclear whether scutellariae can alleviate NAFLD and related pharmacological mechanisms.ObjectFrom the perspective of mitochondrial function regulation,cell and animal experiments were used to investigate the alleviation effect and mechanism of PK on NAFLD.MethodCell experiment: 20 SD male rats were randomly divided into normal control group(normal saline),drug-containing liver mitochondria group(water extract of PK,4 g/kg),continuous intragastric administration for 6 weeks,once a day.Rats in the normal control group were fed with basal diet,and rats in the mitochondria group containing the drug were fed with basal diet.At the end of the experiment,the fresh liver was taken,and the liver mitochondria were obtained by differential centrifugation after homogenization.The normal liver mitochondrial samples and the PK chinensis medicinal liver mitochondria samples were obtained after methanol precipitation.Liver L-02 cells were inoculated into 6-well plates and divided intonormal group,model group,and drug-administered group(mitochondrial samples).Except the normal group,the other groups were induced with liver liter L-02 with 5%medical fat emulsion molding solution.After 24 h,the mitochondria samples(normal mitochondrial group 50 ?g/mL,100 ?g/mL,200 ?g/mL,PK-containing mitochondrial group 50 ?g/mL,100 ?g/mL,200 ?g/mL)were administered to medical fat.The milk-induced liver L-02 cells were further cultured for 24 hours.The lipid droplets in the cells were observed under a microscope,and the cells were used to detect triglyceride(TG),total cholesterol(TC),malondialdehyde(MDA),and superoxide disproportionation.Enzyme(SOD)content.Animal experiments: 63 SD male rats were randomly divided into normal control group(normal saline),model group(normal saline),resveratrol group(positive control,40 mg/kg),and low water extract of raw PK,medium and high dose groups(1,2,4 g / kg),low,medium and high doses of PK extract(1,2,4 g / kg),continuous intragastric administration for 14 weeks,daily One time,except for the normal control group,rats in each group were fed with high-fat diet for 14 weeks to induce NAFLD.Blood was taken from the corner of the eye for 14 weeks.The kits were used to detect serum TG,TC,aspartate aminotransferase(AST),alanine aminotransferase(ALT),high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)levels.After the rats were sacrificed,the organ(liver,spleen,kidney)index was measured,and the levels of TC,TG,and HDL-C in the liver were detected by the kit,and MDA,SOD,and glutathione peroxidase(GSH)in the liver mitochondria were detected.-PX),ATP synthase and respiratory chain complex I,II levels;qPCR detection of UCP-2 and CPT-1 mRNA expression in rat liver;Western blot analysis of rat hepatocyte apoptosis related protein(Caspase-3,Caspase-9,Cytchrome c,Bcl-2,Bax)expression level.ResultsCell experiments: except for the normal group,the other groups produced lipid droplets after induction of liver L-02 cells for 24 h.After administration of mitochondria containing scutellariae,the lipid droplets in the mitochondrial cells of the PK-containing essence were obvious.Decreased,and TC,TG,MDA levels were significantly lower(P < 0.01,P < 0.001),while SOD activity was significantly increased(P < 0.001).Animal experiment: High-fat diet-fed rats were successfully induced withNAFLD.After giving raw and processed PK extracts,the liver index of rats was significantly decreased.Hepatocyte swelling,degeneration,necrosis and inflammatory damage were alleviated.Liver TC,TG decreased significantly(P < 0.05,P < 0.01,P < 0.001)and HDL-C significantly increased(P < 0.01),serum TC,TG,ALT,AST,LDL-C decreased(P < 0.05,P < 0.01).,P < 0.001)and HDL-C were significantly increased(P < 0.05),MDA content in liver mitochondria was significantly decreased(P < 0.05)and SOD,GSH-PX,ATP synthase and respiratory chain complexes I and II were significantly active.Elevated(P < 0.05,P < 0.01,P <0.001),the UCP-2 mRNA was significantly down-regulated and CPT-1 mRNA was up-regulated in the liver,and the expression levels of Caspase-3,Caspase-9 and Bax in the liver were significantly decreased(P < 0.05,P < 0.01,P < 0.001),the expression of Cytchrome c in Bcl-2 and liver mitochondria was significantly increased in the liver(P < 0.05,P < 0.01,P < 0.001).Comparison between the raw and processed PK and the same dose group,the levels of processed PK serum ALT,AST and LDL-C in the serum and blood stasis were significantly lower than those in the raw PK(P < 0.05),and the TC level in the liver of the processed PK stasis was significantly lower than that in the raw PK(P < 0.05),the processed PK HDL-C level of the liver was significantly higher than that of the raw PK(P < 0.01),and the MDA content of the raw PK was significantly lower than that of the processed PK(P < 0.05,P < 0.01),and the raw PK SOD,GSH-The PX level was higher than that of P.chinensis P < 0.05,P < 0.01).The activity of Na+-K+-ATPase and Complex II was significantly higher than that of processed PK(P < 0.01,P < 0.001,P < 0.001).The expression levels of Caspase-3 and Caspase-9 in raw PK were significantly lower than those in processed PK(P <0.05),and the expression levels of Cytchrome c and Bcl-2in processed PK were significantly higher than those in raw PK(P < 0.05,P < 0.001).The expression of Bax in raw PK was significantly lower than that in processed PK(P< 0.001).ConclusionsThe PK-containing liver mitochondria can alleviate the steatosis of liver L-02 cells induced by medical fat emulsion.Both the raw and the processed PK can alleviate the high-fat diet-induced NAFLD by improving mitochondrial function,and the mechanism may be related to enhancing the antioxidant activity of mitochondria.Stress,energy metabolism,fatty acid ?-oxidation and inhibition of hepatocyte apoptosis,and PK can be used to prevent the development of mitochondrial regulators(nutrients)for NAFLD.The effect of processed PK chinensis in reducing serum and liver lipids,increasing mitochondrial energy metabolism and inhibiting mitochondria-mediated hepatocyte apoptosis is better than raw PK.The clinical application of PK and the in-depth development and utilization of resources provide an important scientific basis.