Study On Extraction And Anti-Inflammatory And Analgesic Mechanism Of Total Flavonoids Of Erigeron Breviscapus

Inflammation is a complex immune defense process in which multiple cells and multiple factors participate in the body due to damage factors such as mechanical damage,bacterial or viral infections.It is directly or indirectly related to many diseases.The basic symptoms of inflammation,including redness,heat,swelling,and pain,are one of the most common basic pathological processes in human diseases.When the body is injured or has suffered an injury collectively,it will produce a feeling of pain,which seriously affects physiological activities.Therefore,the discovery of natural medicines with anti-inflammatory and analgesic effects has gradually become a hot work in scientific research.In this experiment,the traditional Chinese medicine Erigeron Breviscapus was selected to explore its antiinflammatory and analgesic activity.In this experiment,the extraction rate of the total flavonoids of Erigeron Breviscapus was used as the screening index.The water extraction and the alcohol extraction were first compared.The results showed that the extraction rate of total flavonoids by the ethanol reflux extraction method was higher than that obtained by the water decoction method.Among them,the breviscapine ethanol The total flavonoid extraction rate of reflux extract was 5.23%.Afterwards,single-factor experiments and orthogonal experiments were carried out to screen and obtain the best extraction process of the total flavonoids of Erigeron Breviscapus,and verify it.The experimental results showed that the optimal solution for extracting total flavonoids from Erigeron Breviscapus was 1:18 g/mL,45%ethanol,soaking time 4 h,and extraction at 80℃ for 1.5 h.At this time,the extraction rate of total flavonoids was 9.73%.The next step is to conduct a methodological investigation experiment on the best extraction process.The effective components of the alcohol extract of Erigeron Breviscapus were analyzed by UPLC,and acetonitrile-0.1%phosphoric acid water was selected for elution.The detection wavelength is 335 nm,and the flow rate is 0.3 mL/min.The elution gradient conditions are 0min:5%acetonitrile,3 min:15%acetonitrile,6 min:15%acetonitrile,8 min:16%acetonitrile,0min:18%acetonitrile.The main components of the alcohol extract of Erigeron Breviscapus are chlorogenic acid,scutellarin,isochlorogenic acid A,isochlorogenic acid C,and the contents are respectively 13.48 ± 0.20 μg/mg、47.60±0.44μg/mg、23.13±0.24μg/mg、22.55±0.27 μg/mg.At the same time,the methodology was investigated,and the results showed that the four components all have a good linear relationship,and their precision,stability,repeatability,and sample recovery are all within the normal range.The results show that the method is feasible.Through DPPH free radical scavenging experiment,reducing power experiment and hydroxyl free radical scavenging experiment,the antioxidant activity of the alcohol extract of Erigeron Breviscapus was done preliminary research.When the concentration is 0.25 mg/mL,the DPPH free radical scavenging rate of the alcohol extract of Erigeron Breviscapus is 92.4%,the hydroxyl radical scavenging rate of the alcohol extract of Erigeron Breviscapus is 94.16%,and the reducing power of the alcohol extract of Erigeron Breviscapus is slightly greater than VC.Through the establishment of inflammation and pain related models,the antiinflammatory and analgesic activity of the alcohol extract of Erigeron Breviscapus was explored.Three models of mouse ear swelling caused by xylene,writhing with acetic acid and pain caused by hot plate were established.The results of the study showed that compared with the model group,the alcohol extract of Erigeron Breviscapus alcohol extract group had an ear swelling inhibition rate of 38.41%,a writhing response inhibition rate of 35.69%,and a pain threshold increase rate 57.21%.The results show that the alcohol extract of Erigeron Breviscapus has certain anti-inflammatory and analgesic effects.Establish a cellular inflammation model of LPS stimulated macrophages to study the antiinflammatory and analgesic mechanism of Erigeron Breviscapus.The results showed that wild baicalin and the effective component group can significantly reduce the NO content in the supernatant of RAW 264.7 cells,and significantly inhibit the expression of iNOS and COX-2 proteins.It shows that Erigeron Breviscapus may inhibit the release of inflammatory factors such as NO produced by macrophages,and down-regulate the expression of iNOS and COX-2 proteins to exert anti-inflammatory and analgesic effects.