The Activity Of Regulating Lipid And Mechanism Study Of Piperlonguminine From Long Pepper

Long Pepper is the grown ear-fruits of which is frequently used in Chinese medicine and Mongolian medicine.That can warm the middle energizer to disperse cold and regulating qi to alleviate pain.It may be not only used in clinic to therapy gastric abscess,disgorging,diarrhea,abdominal pain,toothache and headache.In the traditional Mongolian medicine,Piper Longum L.was used as for treating hyperlipemia and coronary heart disease.Recently,piperlonguminine and it's effective component were studied increasingly,and it's pharmacologic action became a focus for many scholars.After a few year's investigation,our team obtained a kind of chemical compound named as piperlonguminine from Long Pepper,and we found out that the piperlonguminine was the most important compound having the hypolipidemic activity for the first time.In order to study piperlonguminine's effect on accommodating lipid metabolism and anti-artherosclerosis,and to investigate its mechanism of action in molecular level,the chemical,medical science,biological and pharmacological methods were adopted."The following studies were carried out in this thesis.Firstly,the piperlonguminine was prepared using piperineas a starting material,and the structure was determined with high performance liquid chromatogram(HPLC),nuclear magnetic resonance hydrogen spectra(H-NMR) and carbon-spectra(C-NMR).Secondly,we observed piperlonguminine's influence on rat's lipid metabolic disorder using hyperlipemia model,and carried out acute toxicity test study in mouse to confirm piperlongguminine's availability about accommodating lipid metabolism and security.We systematically observed piperlonguminine's effect on HLP and AS rabbits,lipid,NO and apoprotein's contents in serum,antioxidation enzymes activities in serum and tissue.The arteriae aorta,heart and liver's pathomorphology was observed and liver's lipid was determined.Thirdly,we detected piperlonguminine's effect on LDLR,ApoB and HMG-CoAR mRNA expression in HLP rats using semi quantitative RT-PCR technique;and on LOX-1 and VCAM-1 gene expression in HLP and AS rabbits using Real-time PCR.Follow study outcomes were obtained by above-mentioned investigation:1.Piperlonguminine was synthesized successfully by semisynthetic method,and the purity was more than 98%,which not only satisfied pharmacological and toxicological experiment,but also provide good way for producing it in the future.2.Animal studies confirmed that Piperlonguminine may lower significantly TC, TG and LDL-C,and heighten HDL-C in HLP rat dose dependently,and can prevent availably serum lipid to heighten and degrade AI. 3.Toxic reaction was not found in mouse acute toxicity when 5g/kg piperlonguminine was administered one time,which manifest piperlonguminine was little toxic,safe and reliable chemical compound,deserving to further lucubrate.4.Piperlongumnine's effect on serum lipid contents in HLP and AS rabbits was observed dynamically,and found that piperlongumnine can lower TC,TG,LDL-C,and AI and more than positive control Simvastatin in TG;Piperlongumnine can lessen aorta athermatous plaque's areas and intima thickening in rabbits that was induced by high-fat feeds,and relieve the degree of aorta pathology injury and coronary artery stenosis,and had better effect on anti-atherosclerosis.5.Piperlongumnine had heighten AI in HLP rats and AS rabbits,and lower ApoB and ApoB/ApoAl,and had effect on accommodating lipoprotein metabolism.6.Piperlongumnine lessened liver lipid contents in HLP and AS rabbits,and relieved liver adipose degeneration extent in HLP rabbits.Piperlongumnine not only caused lipid's accretion accumulation in liver,but also lowered TC and TG;which indicated initially had preventive and therapeutic effected on adiposis hepatica.7.Piperlongumnine heightened serum NO in AS rabbits,and protected vascular endothelial cell.8.Piperlonguminine enhanced SOD,CAT,GSH-Px activity,reduced MDA contents in HLP and AS animals,elevate antioxidation capability,and decreased free radical and lipid peroxidate damage to blood vessel endothelium.9.LDLR,ApoB,HMG-CoAR gene expression in HLP rat's liver was detected by RT-PCR technique,which indicated that piperlonguminine could accommodate serum lipid through elevating LDLR'gene transcriptional level and lowering ApoB m RNA expression,and had nothing to do with HMG-CoAR m RNA expression.10.Aorta LOX-1,VCAM-1 gene expression in HLP and AS rabbits could be inhibited by piperlonguminine and decrease 42%and 51%respectively comparing with model group,which is possibly a molecule mechanism for piperlonguminine to resist AS.All in all,we found firstly that piperlonguminine can accommodate lipid metabolic disorder and resist AS and its toxicity is very little and it is a kind of safe, reliable chemical compound that can accommodate lipid metabolic disorder and resist AS,which mechanism of action is possibly concerned with elevating liver LDLR and lowering ApoB gene expression,and enhancing antioxidase activity and anti-oxidation ability,and degrading aorta LOX-1 and VCAM-1 gene expression and so on.The outcome provided scientific experiment base for piperlonguminine's further investigation and development.Furthermore,it has important significance for elucidating Chinese and Mongolian drug's molecule mechanism and breakthrough drug's study and exploitation on accommodating lipid metabolism and anti-artherosclerosis.