Development Of Shen Bei Yifei Dripping Pill And Its Effect On Pulmonary Fibrosis Cell Model

Objective:Prepare Shenbei-Yifei Dripping Pills(SYDP)with ginseng radix et rhizoma and fritillariae ussuriensis bulbus as raw materials,optimize the extraction and refining process,select excipients,optimize formulation prescriptions,optimize preparation parameters,establish quality standards for SYDP,and observation of its effect on pulmonary fibrosis cell model.Methods:1.Extraction and purification:Ginseng and Fritillaria were extracted with ethanol as the solvent and refined by adsorption and clarification.The factors affecting the extraction and purification process were investigated.The comprehensive scores of Ginsenosides and Fritillaria B were used as indicators to optimize the extraction process.Parameters,optimization of the refining process with extract yield and component retention;extracts were vacuum dried,crushed through a 120 mesh sieve,and powder properties determined.2.Formulation prescription:The preparation excipients were selected based on the compatibility of the original excipients and the dropping pill forming index as an indicator,to optimize formulation prescriptions.3.Preparation of process parameters:The overall score of each index of the dropping pills forming was used as an index to optimize the small-scale test process parameters,and the best process parameters were determined through the pilot test.4.Quality standard study:Check the formulation indicators such as the appearance of pills and the difference in the weight of pills;TLC for the identification of ginseng and Fritillaria;Determination of ginsenosides Rgi,Re,and Rb1 by high performance liquid chromatography,using acid dyes Method to determine the content of beta-femamin B.5.The influence of pulmonary fibrosis cell model:Human lung adenocarcinoma cell line SPC-A1 induced by transforming growth factor-β1(TGF-β1)was used as a model cell and given different concentrations of SYDP.The expression differences of α-SMA and collagen type 1(Coll)in the model cells before and after administration were compared.Results:1.The optimum extraction process parameters were:70%ethanol extraction 2 times,2 hours each time,solvent dosage 14-fold liquid-material ratio,12-fold liquid-material ratio.2.The purification process parameters are as follows:condensed to the ratio of material to liquid at 1:2,and the temperature is kept at 60℃.Add component B first,and the amount is 7%.Stir every 30 minutes.After 2 hours,add 3.5%of A component,stand for 6 h,filtration,that is to obtain the refined liquid;Purified liquid concentrated and dried at 60℃ for 24 h,to obtain raw material extracts,yield of extract was 10.44%,the raw material was crushed 120 mesh sieve to measure the fluidity,Hygroscopicity and other powder properties.3.Select the PEG4000 and PEG6000(3:1)mixed matrix as the dropping pill matrix and PEG400 as the wetting agent for the raw material and matrix mixing.The formulation of SYDP is:extract 33.33%,PEG 4000 50%,PEG6000 16.67%,PEG400 amount.4.The optimum process parameters are as follows:drip rate 1 drop/s,drop distance 10 cm,dropping temperature 75℃.5.SYDP is a brownish yellow dripping pill with obvious ginseng aroma and completely disintegrated within 10 minutes;TLC identification results show that in the corresponding position with the reference drug chromatography and reference substance chromatography,The chromatograms of the test samples showed the spots or fluorescent spots of the same color,respectively;the average content of benzin B in the 3 batches of samples was determined to be 21.52 μg·g-1 by the acid dye colorimetric method;the content of the 3 ginsenosides was determined by the HPLC method at 20 to 100 Within the range of μg·mL-1,there was a good linear relationship between the content of three ginsenosides and the peak area.The RSD values of precision,reproducibility and stability in the methodological survey were all less than 2%,and the recoveries were 99.40%.The average contents of ginsenoside Rg1,Re,and Rb1 in three batches of samples were 100.96%and 99.96%,which were 0.6716 mg/g,0.5419 mg/g,and 1.0324 mg/g,respectively.6.When TGF-β1 induced SPC-A1 cells for 12 h,the cells became fusiform and spindle-shaped,and the cell gap was slightly larger and the degree of fusion was decreased.After the model cells were given drugs for 24 h,the cell morphology became oval.The interval between cells became smaller;when the dose was 600 μg·mL-1,the expression of α-SMA and Coll was significantly down-regulated.Conclusion:The optimized extraction and refining process of ginseng and Fritillaria are reasonable and feasible.The formulation of dripping pill preparation is reasonable and the preparation process is stable and feasible.The established quality standard has good repeatability and specificity and can be used for the quality of SYDP.Control;The raw materials of this product have certain intervention effects on the morphological changes and partial protein expression of pulmonary fibrosis.