Compare The Chemical Composition And Study The Processes Of Latifolin In Rats Of Dalbergiae Odoriferae Lignum And Heartwood Of Dalbergia Cochinchinensis Pierre Ex Laness

Dalbergiae odoriferae lignum is a dried heartwood of the trunk and roots of Dalbergia odorifera T.Chen,a legume plant.Modern research shows that fragrance mainly contains volatile oil and flavonoids,which have the effects of relaxing blood vessels,increasing coronary flow,antioxidation,anti-inflammatory,antitumor and antibacterial.Dalbergia cochinchinensis Pierre ex Laness and Dalbergia odorifera T.Chen are both plants of the genus Dalbergia,commonly known as“red rosewood”,mainly distributed in Cambodia,Laos,Thailand,Vietnam and other countries.At present,it is artificially planted in Yunnan,Guangxi,Fujian,Hainan and other places in China.The heartwood of Dalbergia cochinchinensis Pierre ex Laness in Thailand is often used as a traditional medicinal material and mainly used in the clinical treatment of blood stasis and cancer.Literature research shows that heartwood of Dalbergia cochinchinensis Pierre ex Laness mainly contains flavonoids.The heartwood of Dalbergia cochinchinensis Pierre ex Laness and Dalbergiae odoriferae lignum is very similar,and it is often adulterated in the Dalbergiae odoriferae lignum.Neoflavonoids are a characteristic component of the genus Dalbergia,and have various biological activities such as antiosteoporosis,antiandrogen,antiinflammatory,antitumor,antiallergic,and antioxidant.Our previous studies showed that latifolin is a neoflavonoids in Dalbergia cochinchinensis Pierre ex Laness and Dalbergiae odoriferae lignum,and has a good protective effect on acute myocardial ischemia and H9c2 cell hypoxia-reoxygenation injury in rats.Based on the phylogenetic theory of“kinship,chemical composition,and pharmacological activity”,in this paper,compare and study the chemical components of Dalbergiae odoriferae lignum and heartwood of Dalbergia cochinchinensis Pierre ex Laness by HPLC,GC-MS and LC-MS methods,molecular docking method was used to study the relationship between neoflavonoids and COX-2 and i NOS enzyme,the pharmacokinetic and metabolomics of latifolin in rats were studied,The main achievements are as follows:I.Study and compare the chemical constituents of Dalbergiae odoriferae lignum and heartwood of Dalbergia cochinchinensis Pierre ex Laness1. Determination of latifolin and seven other chemicals by HPLC: HPLC methods for determining latifolin and seven other chemicals in Dalbergiae odoriferae lignum and the heartwood of Dalbergia cochinchinensis Pierre ex Laness were established,It was found that the contents of 8 constituents in different batches of them were quite different,The percentages of liquiritigenin,luteolin,naringenin,isoliquiritigenin,formononetin,dalbergin,latifolin and pinocembrin are as follows:0.1341%?0.4952%,0.0282%?0.1670%,0.0163%?0.5913%,0.0535%?0.1880%,0.1424%?0.6401%,0.0680%?0.5907%,0.0032%?1.9807%,0.0096%?0.7402%;and only five components were detected in the heartwood of Dalbergia cochinchinensis Pierre ex Laness,and the percentage content of latifolin was0.4736%?2.3815%.HPLC fingerprints and similarity analysis showed that there were significant differences among the 18 batches of Dalbergiae odoriferae lignum,only 11had a similarity value above 0.5,but the similarities of the 5 batches of Dalbergia cochinchinensis Pierre ex Laness were above 0.85;PCA and PLS-DA analysis results show that 18 batches of Dalbergiae odoriferae lignum can establish a better PLS-DA analysis model.2. Using GC-MS method to analyze the volatile components of Dalber giae odoriferae lignum and heartwood of Dalbergia cochinchinensis Pierre e x Laness:A GC-MS analysis method for headspace injection,phenyl alcohol e xtractand volatile oil of Dalbergiae odoriferae lignum and heartwood of Dalber gia cochinchinensis Pierre ex Laness,but there is a great difference between th em;The volatile components of Dalbergiae odoriferae lignum are mainly trans-nerolidol,7-?2,6-dimethyl-hepta-1,5-dienyl?-3,8,8-trimethyl-bicyclo[4.2.0]oct-2-ene and dihydro-3-?2-methyl-2-propenyl?-2,5-furandione,which contain characteristic components such as bisabolene,?±??-bisabolol,farnesene and farnesol;The to p content of headspace injection and phenyl alcohol extract of Dalbergia cochi nchinensis Pierre ex Laness are 3-pyridinemethanol,[1R-?1R*4Z,9S*?-bicyclo7.2.0]undec-4-ene and methyl 4'-methoxy-4,5-methylenedioxybiphenyl-2-carboxylate.3.Identification and analysis chemical components of Dalbergiae odoriferae lignum and the heartwood of Dalbergia cochinchinensis Pierre ex Laness by UPLC-Q-TOF-MS/MS:The UPLC-Q-TOF-MS/MS rapid identification and analysis method of methanol extract from Dalbergiae odoriferae lignum and the heartwood of Dalbergia cochinchinensis Pierre ex Laness was established.A total of 83 constituents were identified from Dalbergiae odoriferae lignum,Including 6 flavones,12 flavanones,18 isoflavones,31 isoflavanones,10 neoflavonoids,4 chalcones,9 isoflavanones,2pterocarpins,3 benzofurans and 6 othercompotents.101 chemical components were identified from the heartwood of Dalbergia cochinchinensis Pierre ex Laness,Including8 flavones,14 flavanones,23 isoflavones,11 isoflavanones,15 neoflavonoids,7chalcones,2 rotenones,5 isoflavanones,3 pterocarpins,4 benzofurans,3phenylpropanoids,2 styrenics and 4 other compotents,of which 85 are known to identify specific structures.There are 54 identical components in the methanol extracts of them,mainly isoflavones,flavones and neoflavonoids.?.Virtual screening of inhibitory activities of neoflavonoids on COX-2 and i NOS enzymesThe molecular docking method was used to virtually screen the 14 neoflav onoids in Dalbergiae odoriferae lignum and the heartwood of Dalbergia cochin chinensis Pierre ex Laness,two enzymes closely related to cardiovascular disea sessuch as ischemic heart disease,COX-2 and i NOS enzymes.It was found th atthe molecular docking scores of latifolin and i NOS enzymes were higher in t he14 neoflavonoids,It scored highest with active site d,only 5-O-methyllatifoli n docked with active site c had a higher score than latifolin,and only 4'-hydr oxy-4-methoxydalbergione and 4,5-dimethoxy--2-hydroxydalbergiquinol docked w ith active site e has a higher score than latifolin;The scores of dockingdalberg iphenols and COX-2 enzyme active point first molecular docking are as follow s:3'-hydroxy-2,4,5-trimethoxydalbergiquinol>2-hydroxy-4,5-dimethoxydalbergiqui nol>5-O-methyllatifolin>latifolin>dalbergiphenol>mimosifoliol,5-OCH₃on the A ring and-OH on the B ring have great influence on the molecular docking of COX-2 enzyme active site a,and the molecular docking score with active site b is:dalbergiphenol>latifolin>2-hydroxy-4,5-dimethoxydalbergiquinol>3'-hy droxy-2,4,5-trimethoxydalbergiquinol>5-O-methyllatifolin,almost the order of t he score of active site a.?.Studying pharmacokinetics of latifolin in rats1.Absorption of latifolin in rats:There was a significant sex difference in the blood drug concentration of latifolin in rats.The pharmacokinetic process in vivo was consistent with the two-compartment model,and the peak time of the blood drug concentration in rats was faster.The peak time of latifolin in rats is faster,the maximum blood concentration time is 30?40min,T_1/2 is also 30?40min,The C_max of the high,middle,and low dose groups were 9.276±1.791,3.06±0.568 and 1.897±0.400mg/L,and the T_max were 40.5±12.778,47.0±7.483,and 28.5±7.806min,respectively;T_1/2 was43.248±11.159,30.361±4.992 and 31.227±7.696min;there are secondary absorption peaks in the curve of the drug time,there is a significant difference between male and female sex,the blood concentration of female rats is significantly higher than that of male rats,and it issignificant statistical differences.The area under the curve was1463.242±123.977mg/L?min and 598.243±46.057 mg/L?min in female and male rats,respectively,with the female significantly higher than the male.2.Study on the distribution of latifolin in rats:Latifolin was distributed in heart,liver,spleen,lung,kidney,brain and tissue of rats,but There is a cert ain difference in the tissue distribution of latifolin in rats.After female rats we re given gavage to the latifolin,The drug concentration in the small intestine a nd stomach was much higher than that in other tissues at 0.5h,1h and 2h tim e points,and the content of latifolin in each tissue at 4h was:brain>fat>small intestine>heart>liver>spleen>lung>kidney>muscle>stomach.After oral administra tion of latifolin to male rats,the concentration ofdrugs in the small intestine a nd stomach 0.5h,1h and 2h three time points isgenerally higher than that in o ther tissues,but the drug concentration in the small intestine tissue decreases r apidly at 1h and 2h.3. Metabolism of latifolin in rats:The metabolites of latifolin in plasma,urine,and feces of rats are different,and prototype drugs and were detected indifferent biological samples Phase?and phase?metabolites.Multiple structures with the same mass spectrometric information as the prototype drug were detected.It is speculated that it may be the isomerization of the L-prototype drug after entering the body,resulting in other configuration components.In additionto phase II metabolism such as sulfate esterification and acetylation,and phase I metabolites such as oxidation,reduction,and demethylation,there are a lot of components that become acid,ketone,and alcohol after oxidation,which maybe related to latifolin contains double bonds and is prone to redox reactions.Literature studies have shown that there is a significant gender difference in CYP450 enzymes in rats.The content of CYP450 enzymes in female rats is 10%to 30%lower than that in male rats.This may be the main reason leading to the significant gender difference in the metabolism of latifolin in rats.4. Excretion of latifolin in rats:The excretion of latifolin in the urine of rats has a gender difference between male and female.There is a significant difference in urinary excretion rate and average cumulative excretion.The urinary excretion rate of rats was significantly faster than that of females,and the average cumulative excretion was significantly greater.The average cumulative excretion rates of latifolin in the urine of female and male rats were 1.21%and 1.43%,respectively,but there was no statistical difference.The average cumulative excretion rates of latifolin in the feces of female and male rats were 17.80%and 14.79%,respectively.The average cumulative excretion rates of female and male rats were different,but there was no significant statistical significance,but the female rats excrete more from feces than male rats.The average cumulative excretion rate of latifolin in rat bile is 1.0940±1.0277%,which varies greatly from individual to individual;the average cumulative excretion rate of prototype in bile of female and male rats is 0.2284±0.1111%,1.9596±0.7756%,There is a significant difference in the average cumulative excretion rate of the prototype in the bile of female and male rats;while the average cumulative excretion and average excretion rate of latifolin in the bile of female and male rats are different,but there is no significant statistical difference.Latifolin is present in the urine,feces and bile in rats,but is mainly excreted through feces,and the prototype excretion rate of feces is higher than 10%.The significant sex difference between the latifolin in the blood concentration of rats may be related to the different excretion rates in male and female rats.?.Metabonomics of latifolin in ratsMetabolomics studies of plasma,urine,feces and bile samples of latifolin in rats.There were found that Plasma,urine,stool and bile samples of rata differences in male and female rats.Different amounts of different metabolites were detected under the positive and negative ion mode.Metabolic pathway analysis found that latifolin may have significant effects on steroid hormone biosynthesis and primary bile acid biosynthesis in rats.The effect of latifolin on steroid hormone biosynthetic pathway may be the reason that it has obvious gender difference in rats.