| OBJECTIVE:By observing the in vitro inhibitory effect of Fuzheng Touxie Jiedu Huayu Decoction and its drug-containing serum in combination with antibiotics on MDRPA,the in vivo protective effect of this decoction on MDRPA-infected elderly pneumonia rats and the effect of immune inflammatory injury,to reveal the anti-MDRPA mechanism of this decoction.METHODS:Using tube dilution method and K-B paper method,to observe the inhibitory effect of Fuzheng Touxie Jiedu Huayu decoction and its drug-containing serum combined with antibiotics on MDRPA in vitro,and the effect of this decoction on the ultrastructural changes of MDRPA under transmission electron microscope.The MDRPA pneumonia model was prepared by oral intubation.The vivo protective effects of this decoction and its combination antibiotics on aging rats,inflammation infection related to different phases,lymphocyte proliferation and lung pathological injury were observed.RESULTS:1 The growth curve of the experimental strain MDRPA was determined.In the tube dilution method,the MIC of the Chinese medicine extract was 0.14 g/ml.The blank serum had no significant antibacterial effect,and the different doses of the medicine-containing serum had a certain antibacterial effect.The MIC of imipenem and imipenem combined with blank serum was 16μg/ml,the MIC of imipenem combined with low dose,middle-dose drug-containing serum was8μg/ml,and the MIC of imipenem combined with large-dose drug-containing serum.4μg/ml.The MIC of ceftazidime and ceftazidime combined with blank serum was 128μg/ml,the MIC of ceftazidime combined with low dose drug-containing serum was 64μg/ml,and the MIC of ceftazidime combined with medium-dose,large-dose drug-containing serum was 32μg/ml.KB paper method,antibacterial effect of antibiotics combined with serum-containing of each group was stronger than antibiotics,antibiotics combined with blank serum.There was no significant difference between antibiotics combined with different doses of drug-containing serum in each group.2 Under transmission electron microscope,the normal MDRPA showed a typical gram-negative bacillus morphology.After intervention with different doses of drug-containing and imipenem,the outer membrane,cytoplasm,and internal structure of the bacteria took a different degree damage.3 The MDRPA pneumonia model was established by the orotracheal intubation method.On the 1st and 3rd day after inoculation,MDRPA was cultured in the lung homogenate of the experimental group,and no MDRPA was cultured in the lung homogenate of the control group(P<0.05).During the observation period,the bacterial load in the lung tissue of the experimental group showed a downward trend with time.At 5th and 7th days after inoculation,MDRPA was not cultured in lung homogenates of the experimental and control groups(P>0.05).Lung pathology is the pathological manifestation of pneumonia.The median lethal dose(LD50)of MDRPA in aged rats was 3.251×108 CFU/ml,and the 95%confidence interval of LD50 was 1.340×108 CFU/ml-5.161×108 CFU/ml.4 The best effective dose was screened.There was a difference between the blank group and the model group,the high-dose group,the medium-dose group,and the low-dose group(P<0.05).The average life time of the blank group and the model group,the high-dose group,and the low-dose group had significant differences(P<0.05).There was no significant difference in the mean life time between the blank group and the medium-dose group(P>0.05).The death protection rates of high-dose group,middle-dose group,and low-dose group were 22.22%,22.22%,and 11.11%,respectively,and the life extension rates were 45.00%,60.00%,and 30.00%,respectively.The blank group(BG)respectively compared with the model group(MG),western medicine group(WMG),and traditional Chinese medicine group(TCMG),the mortality rates were significantly different(P<0.05).The BG respectively compared with Chinese medicine advanced administration group(TCMAG)and Chinese and western medicine treatment group(TCM-WMG),There was no significant difference in the mortality rate(P>0.05).The BG was significantly different from the MG and the TCMG on average life time(P<0.05),and there was no significant difference between the BG and the WMG,the TCMAG,and the TCM-WMG(P>0.05).WMG,TCMG,TCMAG,TCM-WMG,the death protection rate of was 33.33%,22.22%,55.56%,44.44%,and the life extension rate was 56.00%.24.00%,88.00%,72.00%.5 Compared with WBC at different time points and different intervention groups,there was no statistically significant difference between groups at each time point,among groups,BG,MG and each drug intervention group(P>0.05);At different time points,there was no significant difference between the BG,MG,and each drug intervention group in each treatment group at different time points(P>0.05).Comparison of NE%:Vertical comparisons were made among groups in each time point,and there was no significant difference in NE%between different treatment groups at each time point(P>0.05).The NE%in the BG was compared at different time points and there was no significant difference(P>0.05).NE%increased at 2h and 1d after infection in each infected group,and decreased at 3d,5d,and 7d.The NE%of the MG was compared with that of 3d,5d,and 7d respectively on the 1st day,and the difference was statistically significant(P<0.05).NE%showed a downward trend at 3d,5d,and 7d.WMG NE%at 2h,1d were compared with 3d,5d,7d,the difference was statistically significant(P<0.05),NE%decreased at 3d,5d,7d;TCMAG NE%at 2h and 1d were compared with 5d and 7d respectively,the difference was statistically significant(P<0.05).The early administration of traditional Chinese medicine decreased NE%at the 5th and 7th days.TCM-WMG NE%at 2h and 1d were compared with 5d and 7d respectively,the difference was statistically significant(P<0.05).The NE%decreased obviously at 5d and 7d.Comparison of LY%:Vertical comparisons were made among groups in each time point,and there was no significant difference in LY%between different treatment groups at each time point(P>0.05).In BGs,there was no significant difference in LY%at different time points(P>0.05).In different infected groups,LY%showed an upward trend at 3d,5d,and 7d after infection.The LY%of the MGs had an upward trend at 3d,5d,7d,compared with 2h and 1d after infection,but the statistics showed no significant difference(P>0.05).In WMGs,LY%at 2 h,1d were compared with 3 d,5 d and 7 d respectively,the difference was statistically significant(P<0.05).LY%rose at 3 d,5 d and 7 d.In TCMAGs,LY%at 2 h and 1 d was compared with that of 5 days and 7 days respectively,the difference was statistically,significant(P<0.05).,the LY%was significantly increased at 5d,7d.In TCM-WMGs,LY%at 2 h was compared with 5 d and 7 d respectively,the difference was statistically significant(P<0.05).LY%at 1d was compared with 5d,the difference was statistically significant(P<0.05).The LY%was significantly increased at the 5th and 7th days.6 T lymphocyte proliferation assay;WMG,TCMG,TCMAG compared with the BG respectively,the proliferation capacity of T lymphocytes were different(P<0.05),WMG T lymphocyte proliferation capacity was lower than the BG,TCMG,TCMAG.There was no significant difference between the TCM-WMG and the BG(P>0.05).The T-lymphocyte proliferation ability of the TCM-WMG was nearly normal.Compared with other groups,the MG had the most proliferative ability of T lymphocytes(P<0.05);WMG had no significant difference in proliferation ability of T lymphocytes compared with TCMG and TCMAG(P>0.05).The T-lymphocyte proliferation ability of the TCM-WMG was different from that of the WMG,TCMG,and TCMAG(P<0.05),and was stronger than that of the other groups.B lymphocyte proliferation assay:Compared with the BG,the TCMG and the MG respectively had a stronger proliferative capacity of B lymphocytes than the BG(P<0.05);WMG,TCMAG,and TCM-WMG compared with the BG respectivel,there was no significant difference in the proliferation ability of B lymphocytes(P>0.05).Compared with other groups,the proliferation ability of B lymphocytes in the MG was higher than that in other groups(P<0.05);WMG had no significant difference in proliferation ability of B lymphocytes compared with TCMG,TCMAG and TCM-WMG(P>0.05);Compared with the TCMG,the B cell proliferation ability of the TCMAG was lower than that of the TCMG(P<0.05).7 Pulmonary pathology:BG of elderly rats with mild chronic inflammation.Pneumonia was manifested at 2 h after model establishment,inflammation was gradually increased at 6 h,inflammation was most obvious at 1 d and 3 d,and it was patchy and substantively infiltrated.Inflammation became stable at 5 d and 7 d.In the drug intervention group,the inflammation of the TCM-WMG was lighter,followed by the TCMAG,followed by the WMG and TCM.CONCLUSION:1 The tube dilution method and KB paper method study showed that the drug-containing serum combined with antibiotics has a synergistic antibacterial effect.And under the observation of transmission electron microscope,the higher the drug-containing serum concentration of antibiotics,the more severe the damage of MDRPA ultrastructure is.2 The MDRPA pneumonia model was successfully established by the intubation method and the LD50 was determined.The TCMAG and the TCM-WMG can reduce the mortality rate of rats and prolong the survival date,which has a protective effect.3 There was no significant change in WBC among different time points or different treatment groups.The NE%of the MG,WMG,TCMAG and TCM-WMG showed a downward trend at the 5th day of treatment,and the LY%showed an upward trend.4 The TCM-WMG can reduce the proliferation of T and B lymphocytes,correct the immune disorders in the early stage of infection,and make the immune status normal.5 Pathological changes of lung tissue:In each drug treatment group,the TCM-WMG had the best treatment effect,with less inflammation in the lungs,less substantial infiltration,and less damage to alveolar structure. |
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