Based On The Changes Of Fatty Acid Degrading Enzyme Expression To Explore The Molecular Mechanism Of Phlegm Promoting Colorectal

Objective: The high-fat diet is a risk factor to colorectal cancer,meanwhile,in T raditional Chinese Medicine theory,it is lead to Spleen deficiency then generating Phl egm syndrom.This study is to clarify the effect of “Phlegm” in a high-fat diet feedin g colorectal cancer(CRC)mouse model.The molecular mechanism of “Phlegm”promo ting CRC was explored from the expression changes of long-chain fatty acyl-CoA synt hetase(Acyl-CoA Synthetase 1,4,ACSL1 and ACSL4),which playing important roles in fatty acid decomposition.This study may provide some experimental supports for the application of traditional Chinese medicine to prevent and treat CRC.Methods: A CRC model was establishe by a single intraperitoneal injection of A OM(20 mg/kg bw)and 3 cycles of 1%(w/v)DSS in drinking water.High fat diet w as used to induce “Phlegm” and Erchen Decoction(ECD),as the drug counterevidenc e,was applied to reuduce phlegm.A totle of 120 male ICR mice were divided rando mly into 6 groups.The control group: normal diet feeding.CRC group: AOM(oxyge nethane)/DSS(dextran sodium sulfate)treatment was used to induce colorectal cancer.Phlegm group: high-fat diet feeding.Phlegm CRC group: high-fat diet and AOM/DSS treatment.ECD+CRC group: high-fat diet,AOM/DSS treatment,Erchen decoction.E CD group: high-fat diet,and treated with Erchen Decoction.The CRC group,the CR C group and the phlegm CRC group were induced by colorectal cancer using the AO M+DSS “four-step method”.The body weight of the mice was weighed weekly,and t he stool condition and mental state of the mice were observed daily.Mice were sacrif iced 36 weeks later for material collection.The tumor incidence rate of each group of mice was compared.The total cholesterol(TC),Triglyceride(TG),and blood glucose(GLU)in the serum of each group were detected and compared,and liver and viscer al fat were observed.Histopathological changes were observed by HE(Hematoxylin a nd eosin)staining of colorectal tissue.The real-time fluorescence quantitative PCR and automated capillary Western blot were used to detect ACSL1 in visceral fat,liver an d colorectal tissues of each group.ACSL4 mRNA and protein expression levels.Results:(1)The incidence of tumors in each group: the incidence rates of tumo rs in the control group,CRC group,phlegm syndrome group,Phlegm CRC group,H uayu CRC group and Huayu group were 0%,66.7%,5%,92.9%,5.3%,0%.(2)Compared with the control group,the body weight,TC and TG of the Phlegm g roup and the Phlegm CRC group were significantly increased(P<0.05),and the body weight,TC and TG were significantly lower than those of the CRC group and the p hlegm group.(P<0.05).(3)Comparison of HE staining results: In the liver tissue sections under light microsc ope,the mice in the control group had normal structure,no steatosis cells,and no inf lammatory infiltration;the liver tissue structure of the CRC group was basically norma l,but inflammatory infiltration was observed;A large number of diffuse hepatocyte ste atosis was observed in the liver tissue of the mice in the syndrome group.In addition to a large number of diffuse hepatocyte steatosis,a large number of inflammatory cel l infiltration were observed in the liver tissue of the CRC group.The CRC group and the phlegm group mice The pathological morphology of liver tissue was significantly improved,and steatosis vacuolar hepatocytes and inflammatory infiltration were signifi cantly reduced.In the colorectal,the mice in the control group had normal structure;t he CRC group and the CRC group had diffuse hyperemia of the intestinal mucosa,m ucosal epithelial erosion,and a large number of inflammatory cell infiltration;the phle gm group had obvious inflammatory infiltration;the phlegm CRC group and phlegm I n the group,the inflammatory infiltration was significantly reduced after the drug inter vention,and the tissue morphology was significantly improved.In the adipocytes,the normal morphology of the control group was moderate and evenly arranged;the size of the fat cells in the CRC group was normal,and the coronary structure was observ ed,indicating the presence of inflammatory;the adipocytes in the phlegm group show ed an increase,and inflammatory cell infiltration was observed;In the CRC group,th e fat cells showed severe deformation and disorder,and a large number of inflammato ry cell infiltration occurred at the same time.The CRC group of phlegm and phlegm group and the phlegm group and the CRC group became smaller,and the inflammator y infiltration disappeared.(4)The expression of ACSL1 and ACSL4 mRNA and protein in liver tissue,adipose tissue and colorectal tissue: CRC group,phlegm syndrome group and phlegm syndrom e CRC group were significantly higher than the control group(P<0.05).After the inte rvention of Erchen Decoction,the mRNA and protein expressions of ACSL1 and ACS L4 were significantly decreased in the CRC group and the phlegm group(P<0.05).Conclusion:(1)The CRC model induced by the high-fat diet and AOM+DSS “f our-step method” is stable and reliable.(2)Phlegm promotes the formation of CRC,Erchen decoction has a certain preventive effect on CRC.(3)Erchentang may have a preventive effect on the CRC by down-regulating ACSL1 and ACSL4.