| Objective:(1)The quality standard of Mongolian Medicine Jizhaomudaoriji was studied to provide experimental data for the formulation of quality standard of this medicine.(2)To investigate the effect of Jizhaomudaoriji on the mouse model of Streptococcus pneumoniae pneumonia.Methods:(1)According to the Chinese Pharmacopoeia(2020 edition),Inner Mongolia Mongolian Medicine Decoction Pieces Processing standards(2020 edition),Inner Mongolia Mongolian Medicine Preparations 2021 edition of the third volume of the relevant methods of the preparation of safflower,grass,chebule,coptis chinensis microscopic identification.The chemical and chemical identification of sal sal was carried out.Thin layer identification of safflower,artificial bezoar and coptis chinensis was carried out.Aconitine limit test was carried out for aconitine.The contents of hydroxyl safflower yellow A and gallic acid in the preparation were determined by HPLC.Chromatographic conditions for hydroxysafflower yellow pigment A were as follows: column XBridge-C18(250×4.6mm,5μm);The mobile phase was methanol-acetonitrile-0.7% phosphoric acid(26:2:72).The column temperature was 30℃.Flow rate 1ml/min;Detection wavelength 403nm;Chromatographic conditions for gallic acid: Column XBridge-C18(250×4.6mm,5μm);Using methanol-0.1% phosphoric acid(1:99)as mobile phase;The column temperature was 30℃.Flow rate 1ml/min;Wavelength270nm;Also carry out routine inspection of Jizhaomudaoriji;(2)84 SPF BALB/C mice were randomly divided into seven groups,(blank group,model group,The one prescription of Jizhaomudaoriji high-dose,low-dose group(provided by international Mongolian Hospital),The second prescription of Jizhaomudaoriji high-dose,low-dose group(provided by Kulun Mongolian medicine factory),Amoxicillin positive control group),Each group of 12 animals was fed daily and their weight was measured.The blood samples were collected from the orbit of the mice two hours after the last administration.The levels of IL-1 and IL-6 in the serum of the mice in each group were detected by ELISA enzyme-linked Streptococcus pneumoniae assay(Elisa),TNF-a expression levels were measured in lung tissue for histopathology assessment.Results :(1)After microscopic identification,there are the following microscopic features.Safflower: The pollen grains can be round,olive or oval in shape,with a diameter of up to 60μm.They also have three germinal pores and small dentate protrusions on the surface.Corolla,filaments,stigma fragments are common,there are long tubular secretory cells often located near the ducts,about 66μm in diameter,containing yellowish brown to reddish brown secretions.Aconitum: The shape of stone cells is various,from square-like,rectangular to fusiform,the diameter is from 20～133(234)μm,the length is up to 465 μm,the wall is slightly thick,some cells contain brown matter and precipitated powder particles.Terminalia chebula:Lignified thick-walled cells are yellowish or colorless and may be oblong,polygonal or irregular in shape,with one end swollen into a boot.Rhizoma Picrorhiza: Cylindrical,slightly curved,occasionally branched,3-12 cm long,0.3-lcm in diameter.Surface grayish brown to dark brown,scabrous,with dense annular nodes,with slightly elevated bud scars or root scars,distal ends densely covered with dark brown scaly petiole residues.Light body,hard and brittle,easy to break,slightly flat section,light brown to dark brown,wood has 4-10 similar white spot vascular bundle arranged in a ring.(2)Bainaosha is a kind of mineral medicinal material,so it is identified by physical and chemical identification.(3)In the analysis of safflower,artificial bezoar and Corydalis by TLC,the same color spots were found in the chromatographic positions of the samples corresponding to those of the control herbs and the control samples,negative controls did not have this characteristic.(4)The chromatographic results of three batches of radix aconitum showed that there were no or less spots in the samples compared with the control samples.(5)The moisture content of Jizhaomudaoriji(one prescription)is between 5.1% ～ 6.25%;The content of the extract ranged from 20.92% to21.47%,with an average of 21.23%.The moisture content of Jizhaomudaoriji(second prescription)is between 6.25% ～7.5%.The content of the extract ranged from 21.65% to22.57%,with an average of 22.08%.The dissolution time of Jizhaomudaoriji(one prescription)is 41 min,47min,52 min,respectively,Jizhaomudaoriji(second prescription)were43min,45 min and 53 min respectively.In the weight difference test,the weight difference limit of each 10 samples in three batches was within± 10%,and no samples exceeded the weight limit.(6)The concentration of hydroxysafflor yellow a in the two High-performance liquid chromatography showed a good linear relationship in the range of 0.0284～0.8549mg/m L,The regression equation was y=101906x-10393(r=0.998,n=6),the average recoveries were 97.01% and 100.27%,and RSD were 0.75% and 0.75% respectively,and the repeatability,precision and stability were in accordance with relevant regulations.After High-performance liquid chromatography detection,the concentration of gallic acid ranged from 0.2795 to 0.8385 mg/m L,and there was a good Linear independence between the two.The regression equation was y=92362x+1126.9(r=1,n=6),and the average recovery was99.15%,RSD=1.10%.The repeatability,accuracy and stability have reached the corresponding standards;(7)After HE examination,it was found that there were obvious defects in the structure of lung tissue in the model group,a lot of neutrophil appeared in the bronchioles and alveolar spaces,red blood cells exudated,the alveolar wall capillary expanded,and the alveolar space decreased,the neutrophil and histiocytes exudate,some alveoli fuse and the alveolar spaces disappear.ELISA showed that the serum levels of IL-1,IL-6 and TNF-α in the model group were significantly higher than those in the control group after 7 days of Streptococcus pneumoniae treatment,the difference was statistically significant(P<0.05).Compared with the model group,the level of inflammatory factors in the serum of mice in the treated group decreased.So the difference was statistically significant(P<0.05).Conclusion(1)the identification and content determination methods used in this experiment are advanced,simple,reliable,specific,repeatable and stable,the results can provide reference data for making quality standard in Jizhaomudaoriji.(2)Jizhaomudaoriji has anti-inflammatory effects on the model of Streptococcus pneumoniae pneumonia. |
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