Study On The Material Basis And Mechanism Of Ginseng-Clove In Alleviating Obesity With Mice

Objective: In order to find medicinal and edible medicinal materials that can alleviate food-borne obesity,large-scale screening was carried out based on Mortierella alpina.In order to explain the composition and effect of ginseng-clove medicine on alleviating obesity,the spectrum-effect relationship study and the activity evaluation study of weight loss effect were carried out.Finally,the mechanism of leptin-mediated JAK2 / STAT3 pathway affecting appetite and fat synthesis AMPK / m TOR / SREBP pathway regulating key enzymes in fat synthesis to alleviate obesity was studied.Methods:1.Based on the high-yield fatty acid Mortierella alpina,the best extraction method of ginseng-syringa herb pair inhibiting its fat synthesis was screened,the TG content in the enzyme was determined,the morphology was observed by electron microscopy and the fatty acid temperament was analyzed,and the m RNA expression of ACC,ACLY,FAS and ME genes in the fat synthesis pathway was determined by q RT-PCR.2.Based on the HPLC method,the fingerprints of 10 batches of ginseng-clove drug pairs were established,the common components were identified and the contents were determined.Through the model of Mortierella alpina,the TG content contained in the mold and the enzyme activities of ACC,ACLY and FAS were detected.3.By establishing a mouse model of food-borne obesity,Micro-CT scanning analysis was used before and after administration intervention,body weight and calorie intake were measured during the cycle,glucose tolerance and low temperature tolerance were measured at the end of the period,and waist circumference,epididymis and perirenal fat were measured to calculate body fat ratio;the serum lipid four items,FFA,GLU,INS,fat secretion factors LEP and ADP,inflammatory factors IL-6 and TNF-α,and the enzyme activities of FAS,ACC and ACLY in epididymal fat were detected.HE staining sections of epididymal and perirenal fat and liver tissue were observed to evaluate the activity of ginseng-clove drug pair in alleviating obesity.4.The proliferation activity of 3T3-L1 preadipocytes was determined by MTT assay,and the ability of fat accumulation during cell differentiation was determined by oil red O staining.The gene expressions of FAS,ACLY and ACC,LEPR,JAK2,STAT3 and SOCS3 in adipocytes were determined by q RT-PCR.At the same time,Western blot was used to detect the changes of leptin resistance-related proteins LEPR,JAK2,STAT3 and SOCS3 in adipocytes and hypothalamus,as well as the protein expression of lipid synthase-related proteins AMPK,m TOR and SREBP.The protein expression of appetite-related proteins AGRP,NPY,MC4 R and POMC in the hypothalamus was detected to study the mechanism of ginseng-clove drug pair in alleviating obesity.Results:1.Based on the screening of Mortierella alpina,the extraction method with the strongest ability to inhibit lipid synthesis was 100 % ethanol extract with a ratio of ginseng to clove of 1 : 2.After drug intervention,scanning electron microscopy and fatty acid temperament analysis showed that the lipid in the mold was reduced,and the enzyme activity of ACC,ME and other important factors in the lipid synthesis pathway including FAS was inhibited by chemical reagent and enzyme-linked immunosorbent assay.2.Based on the HPLC method,the appropriate chromatographic conditions were successfully established,and the fingerprints of 10 batches of ginseng-syringe medicinal pairs from different producing areas were established.According to the fingerprints,it was found that the similarity of 10 batches of ginseng-syringe medicinal materials was high,and18 common components of gallic acid,chlorogenic acid,ferulic acid,Rg1,Re,quercetin,eugenol,isoeugenol,Rg2,Ro,Rb1,Rd,Rb2,Rb3,Rd,acetyleugenol,isoacetyleugenol and dehydrodiisoeugenol were identified.The ’ spectrum-effect ’ relationship study found that ginseng clove extract inhibited the enzyme activities of ACLY,FAS and ACC in mold,and reduced the accumulation of total lipids in mold.It was found that peaks 5 and 7 contributed greatly to inhibiting enzyme activity and reducing lipid accumulation in M.alpina.3.Successfully established food-borne obesity,after administration intervention,it was found that the intake of calories in obese mice was controlled,and the weight gain was alleviated to some extent.The weight and body fat of waist circumference,epididymal and perirenal fat were significantly lower than those in the model group,and the glucose regulation ability and body temperature regulation ability were significantly improved.Serum detection showed that compared with the model group,the levels of four blood lipids,FFA,GLU,INS,LEP,IL-6 and TNF-α in the serum of mice after administration were significantly decreased,while the ADP content was increased,and the enzyme activities of FAS,ACC and ACLY in epididymal fat were inhibited.HE staining showed that the single volume of epididymal and perirenal fat in mice after administration was significantly smaller than that in the model group,and the lipid droplets in the liver were significantly reduced.4.AEGC intervention was performed during the differentiation of 3T3-L1 preadipocytes into adipocytes,and oil red O staining and semi-quantitative analysis showed that the lipid droplets in the cells were significantly reduced,and the triglyceride content in the cells was reduced.The enzyme activities of ACC,ACLY and FAS were inhibited.The m RNA expression of FAS,ACC,ACLY and SOCS3 was down-regulated and the m RNA expression of LEPR,JAK2 and STAT3 was up-regulated by q RT-PCR.Western blot showed that the protein expression of SOCS3 and p-m TOR in cells and hypothalamus decreased,while the protein expression of LEPR,JAK2,STAT3 and AMPK increased.The protein expression of SREBP in cells was down-regulated,the protein expression of AGRP and NPY in thalamus was down-regulated,and the expression of MC4 R and POMC was upregulated.Conclusion:The 1:2 100 % ethanol extract of ginseng-syringa screened by MA can reduce the synthesis and accumulation of fat in MA and adipocytes,and alleviate food-borne obesity in mice.The mechanism of action is to alleviate the occurrence of obesity through leptin and its receptor-mediated JAJ2 / STAT3 pathway and affecting appetite regulators and fat synthesis factors AMPK / m TOR / SREBP.