The Study On The Relationship Between Leptin And KLF14 In The Formation Of Cholesterol Stones In Mice

Objective:A mouse model of gallbladder cholesterol stones was established by high-fat diet lithogenic method.Exogenous leptin intervention in mice during the establishment of the mouse model.The stone formation,HDL,LDL,TC,Apo-A1,serum leptin levels and hepatic Klf14 expression levels of cholesterol stones in the gallbladder of mice were observed.To investigate the possible relationship between leptin and KLF14 in the formation of cholesterol stones in the mouse gallbladder and the mechanisms linking them.Methods:In this experiment,40 healthy female C57BL/6J mice were selected for the study and all mice were acclimatized and fed for 2 days.The mice were divided into four groups of10 mice each using the random number table method:Basal feed saline control group(G-NS group),basal feed leptin intervention group(G-Leptin group),high-fat feed saline control group(L-NS group),and high-fat feed leptin intervention group(L-Leptin group).Mice in the G-NS and G-Leptin groups were fed basal chow.Mice in the G-NS and G-Leptin groups fed a basal diet.Mice in the L-NS and L-Leptin groups fed a high-fat diet.After 6 weeks of chow feeding,mice in the G-Leptin and L-Leptin groups were given leptin for intraperitoneal injection,and mice in the G-NS and L-NS groups were given saline for intraperitoneal injection.The injections were given every other day for 2 weeks.After all mice were kept for8 weeks,the eyes of the mice were removed for blood sampling,and the mice were executed using the neck dislocation method.Retain bile specimens and liver specimens and record data after testing:(1)The stone formation of bile and gallbladder was observed.Cholecystolithiasis in mice was observed by polarizing microscope after bile smear.(2)Remove the eyeball to take 2m L of blood,let it stand for 2 hours at room temperature and then centrifuge at3500 rpm for 10 min to take the serum.Measure TC,HDL,LDL,Apo-A1,levels in serum samples.Mouse serum leptin level was measured by ELISA.Determination of Klf14 expression levels in liver tissues by quantitative fluorescence q PCR.The results were finally statistically analyzed to draw correlation conclusions.Results:1.Stone forming rate: 0%,40%,60% and 80% in the G-NS,G-Leptin,L-NS and L-Leptin groups,respectively.2.Serum total cholesterol content: G-Leptin group was lower than G-NS group(P <0.01),L-NS group was higher than G-NS group(P < 0.01),L-NS group was higher than L-Leptin group(P < 0.01),L-Leptin group was higher than G-Leptin group(P < 0.01).3.Serum low density lipoprotein content: L-NS group was higher than G-NS group(P <0.01),G-Leptin group was higher than G-NS group(P < 0.05),L-Leptin group was higher than G-Leptin group(P < 0.05).4.Serum high density lipoprotein and apolipoprotein A1 content: L-NS group was lower than G-NS group(P < 0.01),L-Leptin group was lower than L-NS group(P < 0.05),L-Leptin group was lower than G-Leptin group(P < 0.05),G-Leptin group was lower than G-NS group(P < 0.05).5.Serum leptin content: G-Leptin group was higher than G-NS group(P < 0.01),L-Leptin group was higher than L-NS group(P < 0.01),L-Leptin group was higher than G-Leptin group(P < 0.01),L-NS group was higher than G-NS group(P < 0.01).6.Liver Klf14 expression: L-Leptin group was lower than L-NS group(P < 0.01),L-Leptin group was lower than G-Leptin group(P < 0.01),L-NS group was lower than G-NS group(P < 0.01),G-Leptin group was lower than G-NS group(P < 0.05).Conclusion:1.High-fat diet can successfully establish a mouse cholesterol stone model.Both leptin and KLF14 may affect lipid metabolism and participate in the formation of cholesterol stone.The serum LDL content of mice increased after high leptin injection.Serum TC,HDL,Apo-A1 content and liver Klf14 expression level decreased.2.Leptin may reduce the serum Apo-A1 and HDL content by inhibiting the expression of KLF14,leading to abnormal lipid metabolism and increased bilious cholesterol content,thus promoting the formation of gallbladder cholesterol junction.