Study On The Antihypertension Mechanism And Metabolomics Of N~6-(2-Hydroxyethyl) Adenosine In Hypertensive Rats

HEA is one of the active constituents of Ophiocordyceps Sobolifera.It is a natural calcium ion antagonist and adenosine A₁ receptor agonist.Calcium antagonists are commonly used in the prevention and treatment of cardiovascular diseases,but the study on the antihypertensive effect of HEA no reports have been reported yet.In this experiment,the HEA decompression mechanism and metabolic mechanism were studied.From HEA extraction,animal experiment,metabolic analysis,and HEA and HSA binding experiments,a series of studies were carried out using biochemical and molecular biological methods.The main results are as follows:HEA extraction and separation:The polysaccharide is removed by water extraction and alcohol precipitation,and the impurities with different boiling points are removed by distillation.The concentrated liquid is again removed from the macromolecular material by large-empty resin,and the mixture is freeze-dried to obtain a milky white powder,and the HEA is further separated by the liquid phase.Codrycyl adenosine showed that HEA peaked around 10 min and the purity of HEA was over 95%using the standard curve.The antihypertensive effect of HEA on model rats:The SD rats were subjected to hypertension model by intragastric administration of L-NAME 10 mg/kg for three weeks.After two weeks,the systolic blood pressure of the model rats reached 140 mmHg or more,the diastolic blood pressure reached above 90 mmHg,above,HEA 2.5 mg/kg,5 mg/kg and 7.5 mg/kg concentrations were administered,and DPCPX was used to detect HEA on A₁R agonism.The experimental results show that HEA has a hypotensive effect on hypertensive rats.The antihypertensive effect of HEA on SHRs:SHRs were divided into groups,SHR control groups and WKY blank control group were added.Four weeks after intraperitoneal injection,relevant experiments were performed.Blood pressure measurement showed that the low and medium concentration of HEA had an obvious antihypertensive effect,but the low-concentration group had a lower anti-hypertensive effect than the middle-concentration group.Serum biochemical indicators showed that HEA can repair the hypertensive renal injury by lowering blood urea nitrogen/creatinine value.HEA hypotensive mechanism may be through the regulation of related receptors in the MAPK pathway,reduce of ERK1/2 protein expression and increase of eNOS protein expression,while reducing the levels of inflammatory factors IL-6 and TNF-?,WB on rats The hypothesis of the above mechanism was demonstrated in myocardial tissue experiments and ELISA serum experiments.Effect of HEA on intestinal microbes in hypertensive rats:The microbial diversity of SHRs after administration was studied.High-throughput sequencing method studied the effects of HEA on intestinal microbes in hypertensive rats from species diversity,species composition,and sample comparison,and proved that HEA can improve hypertension by increasing the richness of beneficial microorganisms in the intestine.The method of GC-MS was used to study the function of the composition of intestinal microbes in hypertensive rats,to analyze whether the effects of drug administration on the intestinal microbes of hypertension,and the metabolic pathway analysis by differential metabolite screening,to find the differences in the various pathways.This part can deeply study the intrinsic relationship between HEA and intestinal microbes from multiple angles.The binding of HEA and HSA:Through the fluorescence spectroscopy,synchronous fluorescence spectroscopy,three-dimensional fluorescence spectroscopy,and molecular docking methods,the binding process between HEA and human serum albumin was systematically studied.Combined with this experiment,HEA can be effectively explored into the human body and human serum albumin.Combining the process and the auction site,the mechanisms of storage,transportation,and transmission of HEA in vivo are further explored,which provides a basis for the analysis of HEA antihypertensive mechanism.Summary of the article:HEA has a significant antihypertensive effect on hypertensive rats.Zoology and molecular biology experiments show that HEA antihypertensive effect is to activate adenosine A₁ receptor,reduce inflammatory factor content,increase eNOS and reduce ERK1/2 protein expression.The amount of expression,which in turn regulates the MAPK pathway is achieved.The antihypertensive mechanism is also associated with changes in microbial diversity and target metabolic differences.The interaction between HEA and human serum albumin reveals the transport mechanism of HEA in vivo and supply references for the development and utilization of Ophiocordyceps Sobolifera.