| Objectives Experiment 1By observing Gao's Pelvic Inflammatory Prescription No.4 on the general condition of rats with chronic pelvic inflammatory disease,rat uterus changes,morphological changes of rat uterine tissue under light microscope,serum immune related factor content,inflammation related factor protein and To investigate the effect of gene expression,explore the anti-inflammatory mechanism of Gao's Pelvic Inflammatory Prescription No.4 on rats with chronic pelvic inflammatory disease.Experiment 2 Observe the minimum inhibitory concentration of Gao's Pelvic Inflammatory Prescription No.4 on three common pathogens of pelvic inflammatory disease.Methods Experiment 1 Ten of 76 female SD rats were randomly selected as a blank group.The remaining 66 rats were made of animal models of chronic pelvic inflammatory disease using phenol gel chemical burns combined with mechanical injury.After 10 days,6 rats were randomly selected to take uterine tissues to evaluate the success of modeling.The remaining 60 were randomly divided into a model group and a Gao's Pelvic Inflammatory Prescription No.4 high(30g/kg),medium(15g/kg),low(7.5g/kg)dose group,Puling Penyankang granule group(3g/Kg),human placental tissue fluid group(1ml/Kg),10 animals per group.During administration,the general conditions of each group were observed and recorded.Continuous dosing after 20 days,put to death in the rat,macroscopic observation records in the rat pelvic and uterus shape changes,stripping the uterus tissue,weigh and wet weight calculation and swelling degree of the uterus and uterine coefficient,HE staining,the microscopic observation records in the rat uterus tissue morphology change,by carrying out the experiment of enzyme-linked immunosorbent(ELISA)method to detect the rat serum immunoglobulin A(Ig A),immunoglobulin G(Ig G),immunoglobulin M(Ig M),adrenocorticotropic hormone releasing hormone(CRH),adrenocorticotropic hormone(ACTH)and cortisol(CORT)content,Immunohistochemistry was used to detect the expression level of monocyte chemotactic protein-1(MCP-1)protein in rat uterine tissue,and real-time fluorescence quantification(RT-PCR)was used to detect the expression level of MCP-1 gene in rat uterine tissue.The data were analyzed and processed by SPSS 25.0 statistical software.Experiment 2Bacteriostasis experiment to observe the bacteriostasis in vitro of Gao's Pelvic Inflammatory Prescription No.4 on the common pathogenic bacteria of chronic pelvic inflammatory disease: escherichia coli,b hemolytic streptococcus,staphylococcus aureus.The microbroth dilution method of 8×12 well U-shaped aseptic orifice plate was used to conduct the in vitro bacteriostasis experiment of Gao's Pelvic Inflammatory Prescription No.4.10 concentration gradients were diluted with the water decoction of Gao's Pelvic Inflammatory Prescription No.4 to observe the Minimum inhibitory concentration of Gao's Pelvic Inflammatory Prescription No.4 to three pathogenic bacteria.Results Experiment 11.General situation of the rats: the rats in the model group had abnormal mental state,poor diet and drinking water,and the trend of weight gain of the rats was the same.As the number of days of administration increased in each group,the mental state of rats in each group improved,the amount of diet and water increased,and the weight increased.Among them,the high and medium dose group and placenta group improved significantly.2.Changes in rat uterus were observed: the left uterus of rats in the model group was obviously enlarged,hyperemic and deformed,filled with a large amount of effusion,and adhered to the surrounding tissues in a large extent.After administration in each group,the uterus of rats in the high-dose,medium-dose and placental groups weresignificantly improved.3.Uterine swelling degree and uterine coefficient results:compared with the model group,the uterine swelling degree and uterine coefficient of each drug group decreased,with a statistically significant difference(P<0.05),among which,the high,medium dose and placental groups of Gao's Pelvic Inflammatory Prescription No.4 improved significantly.4.Morphological changes of rat uterine tissue were observed under light microscope: the rat uterine cavity epithelium in the model group was seriously damaged,a large number of inflammatory cells were infiltrated,muscle layer was damaged,and intertissue adhesion was serious.After administration,all groups showed improvement,among which the high,medium and placental groups showed significant improvement.5.The contents of Ig A,Ig G,Ig M,CRH,ACTH and CORT in the serum of rats were observed by ELISA method.Compared with the model group,the contents of Ig A,Ig G and Ig M in the serum of rats in the high-dose,medium-dose and low-dose groups of Gao's Pelvic Inflammatory Prescription No.4 were increased,while the contents of CRH,ACTH and CORT were decreased,and the difference was statistically significant(P<0.05).6.Immunohistochemistry was used to detect the expression level of MCP-1 protein in rat uterine tissue.Compared with the model group,the expression level of MCP-1 protein in the high-dose,medium-dose and low-dose groups of Gao's Pelvic Inflammatory Prescription No.4 decreased,and the difference was statistically significant(P<0.05).7.The expression level of MCP-1 gene in the rat uterine tissue was detected by RT-PCR.Compared with the model group,the gene expression level of the high,medium and low dose groups of Gao's Pelvic Inflammatory Prescription No.4 decreased,and the difference was statistically significant(P<0.05).Experiment 2 Gao's Pelvic Inflammatory Prescription No.4 has a significant inhibitory effect on the three most common pathogenic bacteria of chronic pelvic inflammatory disease,e.coli,b hemolytic streptococcus and staphylococcus aureus,with the minimum bacteriostatic concentrations of 62.5mg/ml,125mg/ml and 62.5mg/ml,respectively.Conclusions Experiment 1 Gao's Pelvic Inflammatory Prescription No.4 can effectively inhibit the inflammatory response of Chronic Pelvic Inflammatory Dsease in SD rats,improve the inflammatory symptoms of the pelvis and uterus,increase the content of Ig A,Ig G,Ig M in the serum,improve the body's humoral immunity,reduce the CRH,ACTH and CORT levels,play an immunomodulatory role,significantly reduce serum MCP-1content,MCP-1 protein expression levels and gene expression levels,and play an antiinflammatory effect.Experiment 2 Gao's Pelvic Inflammatory Prescription No.4 can effectively inhibit three common pathogenic bacteria of chronic pelvic inflammatory disease in vitro: Escherichia coli,Staphylococcus aureus,type B hemolytic streptococcus.Figure 4;Table 11;Reference 102. |
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