Effect Of Qutanhuoxue Decoction On PPARα/CPT-1 Pathway And Insulin Resistance In NAFLD Rats

Objective:In this experiment,the rat model of nonalcoholic fatty liver disease was reproduced by feeding high-fat diet,and the intervention of Qutanhuoxue decoction was used to detect the relevant indicators,and then observed and summarized the effects of the Qutanhuoxue decoction on the PPARα/CPT-1 pathway and insulin resistance in NAFLD rats,and explored its mechanism of treating NAFLD from the perspective of lipid metabolism.To provide the basis for the clinical practice experience and academic theory inheritance of famous veteran TCM.Methods:1.Grouping,modeling and drug delivery(1)32 SPF male SD rats were randomly divided into group C(a total of 6 in the normal group)and group model building(a total of 26).The rats in normal group chose to be fed with sufficient normal feed,group model building Rats were fed sufficient high-fat diet.Except for the different diets,the other conditions were the same in normal group and model building.Rats in each group moved freely,ate and drank water.The modeling process lasts for 8 weeks.(2)8 weeks after modeling,randomly select 2 rats from the model building group to verify whether the modeling is successful,fast for 12 hours,weigh,anesthetize,dissect and separate the rat liver,and wash the excess blood with normal saline,Absorbent paper to dry the excess water,take the left liver lobe tissue for hematoxylin-eosin(HE)staining,get the liver pathological results,and compare the NAS points to confirm the successful replication of the NAFLD rat model.The remaining NAFLD rats(24 rats)were randomly divided into groups M(6 rats in the model group),group D(6 rats in the low-dose group of Qutanhuoxue decoction),and group Z(6 rats in the middle-dose group of Qutanhuoxue decoction),H group(6 mice in the high-dose group of the Qutanhuoxue decoction).(3)Feed supply remains unchanged after grouping.Rats in group C and group M were given a dose of 10 mL·kg-1·d-1 with medical grade sterile 9%NaCl injection;rats in groups D,Z,and H were given 6,12,16 g· The dose of kg-1·d-1 was given to the stomach with the liquid of the Eliminating Phlegm and Blood Circulation Recipe.The total duration of this process is 4 weeks.2.Specimen collection and processing(1)At the 4th weekend of the intervention,fast for 12 hours,weigh,anesthetize,open the abdominal cavity of the rat,perform abdominal aortic puncture with a disposable blood collection needle,collect about 5 mL of arterial blood from the negative pressure blood collection tube,and leave it for 4 hours before centrifugation,Keep the upper clear serum,store it in the refrigerator at-80℃for later use.(2)Sacrifice the rat,dissect the rat,extract the liver,and weigh the mass.The left lobe of the liver was fixed in neutral formalin and stained with HE;the right lobe of the liver was also taken about 1.0cm×1.0cm×1.0cm for frozen section and stained with Oil Red O;the remaining liver was placed in a cryotube.Store in-80℃ refrigerator.3.Compare the effects of Qutanhuoxue decoction on liver pathological changes in rats with non-alcoholic fatty liver disease HE and Oil Red O staining were used to observe the effect of Qutanhuoxue decoction on liver pathology in rats with non-alcoholic fatty liver disease.4.Compare the effects of Qutanhuoxue decoction on related serum biochemical indexes in rats with non-alcoholic fatty liver disease(1)The kit detects rat serum ALT,AST,TG,TC,LDL-C,HDL-C,FBG content.(2)ELISA method to detect the content of FINS in rat serum.(3)Calculate HOMA-IR.5.Compare the effects of Qutanhuoxue decoction on the expression of PPARa,CPT-1,IRS-1 mRNA in rats with non-alcoholic fatty liver disease Real-time PCR method was used to detect the expression of PPARa,CPT-1 and IRS-1 mRNA in liver tissues of rats with non-alcoholic fatty liver disease.6.Compare the effects of Qutanhuoxue decoction on the protein expression of PPARα,CPT-1,IRS-1 in rats with non-alcoholic fatty liver disease Western blotting method was used to detect the expression of PPARα,CPT-1,IRS-1 protein in rats with non-alcoholic fatty liver disease by Qutanhuoxue decoction.Results:1.Qutanhuoxue decoction can improve the general state of non-alcoholic fatty liver disease rats such as body shape,coat color and stool.2.Qutanhuoxue decoction can improve the degree of liver steatosis and inflammation in rats with non-alcoholic fatty liver disease.3.Qutanhuoxue decoction can significantly reduce ALT,AST,TC,TG,LDL-C in rats with non-alcoholic fatty liver disease,increase HDL-C,significantly reduce FBG,FINS,HOMA-IR,promote lipid metabolism and improve Insulin resistance.4.Qutanhuoxue decoction can up-regulate the expression of PPARα,CPT-1,IRS-1 protein in the liver tissue of the rat model of non-alcoholic fatty liver disease.Conclusion:1.The structure of the liver lobules in the liver tissue of rats in the M’group is disordered,with circular lipid droplets of different sizes visible,and the NAS score is ≥4,which proves that the modeling is successful;2.Qutanhuoxue decoction can improve liver tissue degeneration and lipid deposition;among them,the high-dose group is better;3.Qutanhuoxue decoction improve serological indicators,among which the high-dose group is better;4.Qutanhuoxue decoction can up-regulate the expression of PPARα,CPT-1 and IRS-1,among which the high-dose group is better;5.Qutanhuoxue decoction can treat non-alcoholic fatty liver disease.Its mechanism of action may be the same as that of Qutanhuoxue decoction which activates the PPARα/CPT-1 pathway and up-regulates the expression of IRS-1,thereby improving liver lipid metabolism and IR.Reduce the degree of liver steatosis and inflammation,thereby treating non-alcoholic fatty liver disease.