Puerarin Hydrogel Attenuates Hyperhomocysteinemia In Mice Through Chronopharmacokinetics By Interacting With Rev-erbα

Puerarin is one of the main active ingredients in Pueraria lobata,a common and traditional Chinese medicine.In addition to its protective effect on the cardiovascular system and liver,puerarin has many medicinal values.It has been widely concerned by researchers,but due to the puerarin The water solubility of the element is not particularly good.The factors such as low oral bioavailability,low membrane permeability,and short half-life limit the use of oral preparations,so the common dosage forms of puerarin are injections,and puerarin injections are used to treat Adjuvant treatment of coronary heart disease,angina pectoris,myocardial infarction and other common diseases has a wide range of clinical applications.But common puerarin injections all add propylene glycol as a co-solvent,although propylene glycol as an additive has a high safety index.But the cost of mass production increases,too much use will also have certain side effects.Because of its structural specificity,puerarin can form a hydrogel by simple operation without chemical modification.Based on the above reasons,we studied whether the puerarin hydrogel injection can be used as an injection for the treatment of diseases.We chose 1%puerarin hydrogel to treat hyperhomocysteinemia.The main research content methods and experimental results of this article are as follows:(1)First,we selected a model of hyperhomocysteinemia in cardiovascular diseases to model mice,and tested whether the modeling was successful.(2)Make puerarin into a 1%hydrogel,give a gradient dose of puerarin hydrogel for one week to treat hyperhomocysteinemia mice to detect various physiological indexes and related gene expression,and related genes Protein levels.(3)The optimal concentration for comprehensive evaluation of drug efficacy was selected at a dose of 200 mg/kg,and mice with high homocysteine at different times(ZT10/ZT22)were injected intraperitoneally with puerarin hydrogel solution.The levels of homocysteine and triglycerides,as well as related RNA and protein in the blood and liver of rats were used to determine the difference in the therapeutic effect of puerarin hydrogel injection at different times.While monitoring the metabolic curve of puerarin hydrogel in vivo at different times,the puerarin hydrogel was intraperitoneally injected into mice at different time points(ZT10/ZT22)to eliminate the factors causing differences in absorption effects at different times.Collect blood at a certain time to measure the content of puerarin.(4)Test whether puerarin is a receptor for Rev-erba by co-expressing chimeric receptors,and detect the protein by adding puerarin 24h after transfection of the promoter and/or high expression knockdown plasmid to cells,RNA and fluorescent reporter genes to verify the mechanism of action of puerarin on cells.The above experiments found that the solubility of puerarin increased sharply after heating and boiling,and cooled after boiling.The solution was all liquid in about 20 minutes.The mice were given intraperitoneal injection of puerarin hydrogel solution for one week.The liver and plasma homocysteine and The value of triglycerides decreases,and the levels of RNA and protein increase.In the data,the change of ZT10 is more obvious than that of ZT22,and the metabolic curve of puerarin administered to mice at different times has no significant difference.Treated with puerarin at the cellular level,the levels of related genes increased.After Rev-erba knockdown,there was no significant difference in these genes between the control group and the administration group.The fluorescent reporter gene is concentration-dependent.Conclusion:In this study,it was found that the puerarin hydrogel dosage form can be used for injection administration.In the mechanism,puerarin induced the expression of Bhmt,Cbs and Cth(three enzymes involved in homocysteine catabolism and the target of Rev-erba),which alleviated methionine-induced hypercysteineemia in mice.In addition,because the rhythm of Rev-erba expresses it as a drug target,there are certain differences in drug efficacy at different drug administration times.In the study,it was found that compared with ZT22 administration,puerarin administration of ZT10 can produce stronger The pharmacological effect is consistent with the antagonistic effect.Based on the luciferase reporter gene,Gal4 co-transfection and target gene expression analysis,it was determined that puerarin is an antagonist of Rev-erba.Overall,this study demonstrates that the temporal pharmacological targeting effect of puerarin on Rev-erba can reduce hyperhomocysteinemia in mice.