Intervention And Clinical Efficacy Of Baixia Decoction On Pseudomonas Aeruginosa Infection

ObjectiveIn this study,the effect of Baixia Decoction on the inhibition of Pseudomonas aeruginosa and its minimal inhibitory concentration were studied in vitro;Through animal experiments to understand the effect of Baixia Decoction in inhibiting Pseudomonas aeruginosa in vivo;Observation of the effect of Baixia Decoction on biofilm formation and structure of Pseudomonas aeruginosa by biofilm experiment;Observing the effect of Baixia Decoction on the treatment of Pseudomonas aeruginosa infection of extremities wounds through clinical research.Methods?1?In vitro bacteriostasis experiment of Baixia Decoction against Pseudomonas aeruginosa,determination of the antibacterial effect of Baixia Decoction on Pseudomonas aeruginosa with medicinal media;determination of antibacterial effect of Baixia Decoction against Pseudomonas aeruginosa by K-B method;determination of the MIC of Baixia Decoction by diploid dilution.?2?Interventional effect of Baixia Decoction on Pseudomonas aeruginosa infection in SD rats,establishment of an acute rat model of Pseudomonas aeruginosa infection in the back of SD rats,wound were wet dressed with BaiXia Decoction,and compared with gentamicin,observe changes in wound area on the first,third,sixth,ninth and 14th days of the experiment,observing the bacteria content in wound granulation,pathological section observation of macrophage and inflammatory cells in granulation tissue.?3?Effect of BaixiaDecoctiononbiofilmformationofPseudomonasaeruginosa,determination of MIC of Baixia Decoction by agar plate dilution method,Baixia Decoction gradient concentration works on Pseudomonas aeruginosa biofilm,crystal violet staining then measures the A value at 650 nm,determination of SMIC₅₀and SMIC₈₀values of Baixia Decoction to evaluate the effect of Baixia Decoction on Pseudomonas aeruginosa biofilms;at the 6th,1st,3rd,and 7th days of the experiment,the effect of Baixia Decoction on the formation and structure of Pseudomonas aeruginosa biofilms was observed and compared with gentamicin and control groups.?4?Clinical observation on the effect of Baixia Decoction on Pseudomonas aeruginosa infection,patients with defects in the wound surface and Pseudomonas aeruginosa infection were selected as observing subjects,60 patients were evenly divided into Baixia Decoction group and gentamicin group,the treatment group was given Baixia decoction soaking daily,and the control group was given conventional dressing gentamycin wet for 14days,observe changes in wound exudation,granulation growth,wound area,inflammatory indicators,and symptom scores of TCM syndromes.Results?1?The results of the in vitro antibacterial effect of Baixia Decoction against Pseudomonas aeruginosa showed that Pulsatilla chinensis,Prunella vulgaris,and Baixia Decoction all had significant inhibitory effects on Pseudomonas aeruginosa.The antibacterial effect was ranked from the largest to the smallest,BaiXia Decoction>Prunella vulgaris>Pulsatilla Chinensis.Kruskal-Wallis test,Chi-Square=25.818,P<0.05,the difference between the groups was statistically significant.The MIC of Pulsatilla chinensis against Pseudomonas aeruginosa is 1:8,the MIC of Prunella vulgaris against Pseudomonas aeruginosa is 1:16,and the MIC of Baixia Decoction against Pseudomonas aeruginosa is 1:32.?2?As a result of the intervention of Baixia Decoction on the infection of Pseudomonas aeruginosa in SD rats,The Inflammatory swelling gradually faded on the first 1-3 days.The pus gradually increased on days 1-6 and then gradually decreased.The pus disappeared on the 9th to the 14th day;the granulation tissue of the wound began to grow with the increase of pus in the early stage,and the granulation tissue of the wound on the 6th day began more.The growth has been obvious,the wounds have become lighter,and the area has decreased.On the 9th to 14th days,the granulation tissue covered the wound surface and the area has further shrunk.Compared with the gentamicin group and the control group,the difference is significant?P<0.01?.On the 14th day,some tissue blocks were taken at the center and edge of the wound surface for pathological section observation under an optical microscope,the number of macrophages was greater in the Baixia Decoction group;inflammatory cells such as lymphoid and neutrophils were more visible in the gentamicin group,and a few macrophages were seen;a large number of necrotic tissues and inflammatory cells were seen in the control group.The bacterial content in wound granulation was significantly decreased in Baixia Decoction group and gentamycin group.On the 5th day after modeling,the bacterial content in granulation of Baixia Decoction and gentamicin group was significantly higher than that of control group,differences?P<0.01?,on the9th day after modeling,the bacterial content in the granulation of Baixia Decoction and gentamicin group was significantly reduced,and there was no significant difference between the two groups?P>0.05?.?3?The effect of Baixia Decoction on the biofilm formation and structure of Pseudomonas aeruginosa was determined by the agar plate dilution method.The MIC of Baixia Decoction was 64 mg/ml,and the concentration of Baixia Decoction acted on the Pseudomonas aeruginosa biofilm.Crystal violet stainingwas used to measure the A value at 650nm.The SMIC₅₀and SMIC₈₀values of Baixia Decoction were 512mg/ml and 128mg/ml,respectively.The gentamicin SMIC₅₀and SMIC₈₀values were 64?g/ml and 8?g/ml,respectively.At the sixth hour,optical microscope showed that the bacteria in the blank group was in the initial stage of adhesion,and the bacterial adhesion in the Baixia Decoction group and the gentamicin group was reduced,but it was still visible;at the first day,optical microscope showed preliminary formation of micro-colony colonies,while micro-colonies in the Baixia Decoction and gentamicin groups were not completely formed;at the third day,optical microscope showed the control group can be seen a large number of bacteria were fused together,the biofilm had been initially formed,and no biofilm was formed in the Baixia Decoction group and the gentamicin group;at the seventh day,Scanning electron microscopy showed that a complete biofilm was formed in the control group,and the biofilm was destroyed in both the Baixia Decoction group and the gentamicin group.?4?The clinical curative effect of Baixia Decoction on Pseudomonas aeruginosa infection in wounds was observed,before treatment,there are no statistical difference about age-rank?P>0.05?and gender?P>0.05?through chi-square test.The area of wounds,syndrome scores,and inflammatory indexes were also analysised by statistical t-test,the P values were all>0.05,there was no statistical difference between the two groups,and they were comparable.The total clinical efficacy after two weeks of treatment was statistically different by non-parametric test P<0.05;after two weeks,the wound reduction rate in the two groups was not statistically different by t-test P>0.05;the main symptoms of TCM syndromes before and after treatment in both groups were analyzed by paired t-test,P<0.01,with statistical difference;the scores of TCM syndromes before and after treatment in the two groups were compared by paired t-test,P<0.01,with statistical difference;after two weeks,the reduction rate of the main symptom score of TCM syndromes was statistically significant after t-test,P<0.05;the reduction rate of TCM syndrome scores after two weeks was statistically significant by t-test,P<0.05;after two weeks,the WBC,NE%and CRP in the two groups were compared by paired t-test,P<0.01,there was statistical difference between the two groups,through intra-group comparison and comparison between groups,the treatment group reduced WBC,NE%and CRP content better than control group;the area of wounds reduced statistically significant?P<0.05?at each time points after treatment intra-group compared with the area before treatment,differences in wound area between the two groups were compared on the 3rd,6th,and 9th day P>0.05,and P<0.05 on the 14th day,there were differences in the later period;the rate of wound healing at two different time points in both groups was slower in both groups?P<0.05?in the early stage and faster in the later stage?P<0.05?,The early experimental group was faster?P<0.05?,and the intermediate and late two groups were equivalent?P>0.05?.Conclusion?1?Pulsatilla Chinensis,Prunella vulgaris and BaiXia Decoction all have antimicrobial effectsagainst Pseudomonas aeruginosa.The antibacterial effects are ranked in descending order:BaiXia Decoction>Prunella vulgaris>Pulsatilla Chinensis.?2?Baixia Decoction can promote wound healing and inhibit the growth of Pseudomonas aeruginosa in the wounds of SD rats infected with Pseudomonas aeruginosa.?3?Baixia Decoction inhibits the early adhesion growth of Pseudomonas aeruginosa to form biofilms and destroy the formed mature biofilm.?4?Baixia Decoction can inhibit the growth of Pseudomonas aeruginosa in wounds,promote the wound healing of patients,and improve the symptoms of traditional Chinese medicine.