| Objective:1.To observe the influence of cake-separated moxibustion with different transdermal enhancers on the blood lipid and liver lipid levels as well as the changes of liver tissue structure and serum lipid metabolism-related enzymes in the rabbits with hyperlipidemia(HLP);2.To observe the influence of cake-separated moxibustion with different transdermal enhancers on the expressions of Leptin/JAK2/STAT3 pathway-related factors,so as to explore the lipid-regulating mechanism of cake-separated moxibustion and the synergistic mechanisms of transdermal enhancer for cake-separated moxibustion.Methods:A total of 40 purebred New Zealand rabbits were selected,from which 8 were randomly selected as normal group(A)fed by common fodders while the remaining rabbits were fed by high-fat fodders for 12 weeks to establish HLP models which were randomized into model group(B),transdermal enhancer-free group(C)(hyperlipidemia model+water as solvent for cake-separated moxibustion),Azone group(D)(hyperlipidemia model+Azone Solution for cake-separated moxibustion).and Borneol group(E)(hyperlipidemia model+ Borneol Solution for cake-separated moxibustion)after successful molding.After 4-week intervention,the rabbits were anesthetized for blood sampling,and then sacrificed to collect the liver tissues for detection.Enzymic method was used to detect serum and liver total cholesterol(TC)and triglyceride(TG)levels,while direct one-step method to detect high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)levels,and transmission turbidimetry to detect the liver tissue apolipoprotein A-1(ApoA-1)and apolipoprotein B(Apo-b)levels.Liver tissue HE staining was performed to observe the morphology and structure.Enzyme-linked immunosorbent assay(ELISA)was applied to detect the serum Leptin,HSL,CYP7A1,and HMG-CoA reductase levels.Immunofluorescence quantitative polymerase chain reaction(PCR)and Western-Blotting were used to detect the expressions of Leptin/JAk2/STAT3 pathway-related factors and proteins.Results:1.In normal group,the hepatocytes were normal in morphology and structure,clear in borders,and well-aligned in arrangement,with round nucleus and obvious nucleolus,without lipid degeneration;in model group,the hepatocytes were swollen into round shape,the cytoplasm was loose and slightly stained,and the arrangement was disordered,accompanied by mild watery degeneration of the hepatocytes,and the hepatic sinusoid was compressed and narrowed,with large amount of droplets noted in the liver;in transdermal enhancer-free group,the hepatocytes were slightly enlarged,the cytoplasm was loose and slightly stained,and the arrangement was disordered,without hepatocytic degeneration,hepatic sinusoid was basically normal,and the intrahepatic lipid droplets were notably less than those in model group;in Azone group and Borneol group,the hepatocytes were basically normal in size and morphology,normal in cytoplasm staining,and well-aligned in arrangement,without hepatocytic degeneration,hepatic sinusoid was normal,and the intrahepatic lipid droplets were obviously less than those in model group and less than those in transdermal enhancer-free group;in Borneol group,the intrahepatic lipid droplets were slightly less than those in Azone group.2.Relative to normal group,model group was markedly higher in serum and liver tissue TC,TG,and LDL-C levels as well as liver tissue Apo-b and notably lower in serum and liver tissue HDL-C level and liver tissue ApoA-1 level(P<0.05 or P<0.01);relative to model group,relative to model group,transdermal enhancer-free group,Azone group,Borneol group were notably lower in serum and liver tissue TC,TG,LDL-C and Apo-b levels and markedly higher in liver tissue HDL-C and ApoA-1 levels(P<0.05 or P<0.01);relative to transdermal enhancer-free group,Azone group was significantly lower in serum and liver tissue TC and TG levels and apparently higher in liver tissue ApoA-1 level(P<0.05 or P<0.01),while Borneol group was notably lower in serum and liver tissue TC,TG,LDL-C and Apo-b levels and prominently lower in HDL-C and ApoA-1 levels(P<0.05 or P<0.01);relative to Azone group,Borneol group was markedly lower in serum TC,TG and LDL-C levels as well as liver tissue TG,LDL-C and Apo-b levels and notably higher in HDL-C level(P<0.05 or P<0.01).3.Relative to normal group,model group was markedly higher in serum HSL and HMG-CoA reductase level and significantly lower in CYP7A1 level(P<0.05 or P<0.01);relative to model group,transdermal enhancer-free group,Azone group and Borneol group were serum significantly higher in HSL and CYP7A1 levels and prominently lower in HMG-CoA reductase level(P<0.01);relative to transdermal enhancer-free group,Azone group and Borneol group were prominently higher in HSL and CYP7A1 levels and notably lower in HMG-CoA reductase level(P<0.05 or P<0.01);relative to Azone group,Borneol group was significantly higher in the increase range of HSL and CYP7A1levels(P<0.05 or P<0.01).but there was insignificant difference between two groups in the influence on HMG-CoA reductase level(P>0.05).4.Serum Leptin ELISA results:Relative to normal group,model group was markedly lower in Leptin level(P<0.01);relative to model group,transdermal enhancer-free group,Azone group,and Borneol group were notably higher in Leptin level(P<0.01);relative to transdermal enhancer-free group,Azone group and Borneol group were significantly higher in Leptin level(P<0.05 or P<0.01),but no difference was noted between Azone group and Borneol group in Leptin level.Liver tissue PCR results:Relative to normal group,model group was markedly lower in the expressions of Leptin,JAK2 and STAT3 mRNA(P<0.01);relative to model group,transdermal enhancer-free group,Azone group and Borneol group were significantly higher in the expressions of Leptin,Leptin Receptor,JAK2 and STAT3 mRNA(P<0.01);relative to transdermal enhancer-free group,Azone group and Borneol group were notably higher in the expressions of Leptin,Leptin Receptor,JAK2 and STAT3 mRNA(P<0.01);relative to Azone group,Borneol group was notably higher in the expressions of Leptin,Leptin Receptor,JAK2 and STAT3 mRNA(P<0.05 or P<0.01).The immunohistochemical test and western-blotting detection tendency was consistent with quantitative PCR results.In addidition,the immunohistochemical test and western-blotting showed that the tendency of P-JAK2 and P-STAT3 was consistent with that of JAK2 and STAT3.Conclusion:1.Cake-separated moxibustion has favorable effect in regulating the rabbits'blood and liver lipid,and protect and repair the liver tissues,for which adding Azone and Borneol as transdermal enhancers will provide better efficacy.2.Cake-separated moxibustion may exerts its lipid-regulating effect by influencing lipid metabolism-related enzymes including HSL,CYP7A1 and HMG-CoA reductase,and Azone and Borneol may play their lipid-regulating synergistic mechanism by impacting above 3 enzymes3.Cake-separated moxibustion can activate Leptin-mediated JAK2/STAT3 lipid-regulating pathway and upregulate the expressions of relevant factors,for which adding Azone and Borneol can more significantly activate this pathway,which may be one of the lipid-regulating mechanisms of cake-separated moxibustion and the synergistic mechanisms of Azone and Borneol as transdermal enhancers for cake-separated moxibustion. |
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