| Objectives By observing the effects of the Gao's Prescription No.3 on behavior,pathology,inflammation related factors and Nuclear Factor-?B(NF-?B)in chronic pelvic inflammatory model rats,to evaluate the curative effect of the Gao's Prescription No.3 in the treatment of CPID(Chronic Pelvic Inflammatory Disease,CPID),to discuss the mechanism of action in the treatment of CPID and to observe the effect on reproductive function.Methods 1.Anti-inflammatory experiments: 60 female Kun Ming mices were randomly divided into 6 groups,10 in each group,the blank group,the Gao's Prescription No.3 high,medium and low dose group,Pu Ling group and Human placental tissue fluid group,7 days to take medicine.In 0.05 ml xylene reagent cause inflammation,remove the double ear in mice,and calculate auricle swelling degree and the inhibition rate.2.Pharmacodynamics and reproductive experiments: 119 female SD rats aged 12 weeks were randomly divided into two groups,pharmacodynamics group(10 rats in each group)and reproductive function group(7rats in each group).Except the blank group,all the other rats were modeled by mechanical injury combined with chemical burn(10d).The rats were divided into blank group,model group,Pu Ling group and Human Placental Tissue Histosolution group,except for the Human placental tissue fluid group,which was administered intramuscularly,the remaining groups were administered by intragastric administration,once a day,then treatment for 20 days.(1)Pharmacodynamics group:To observe the behavior,morphology and histopathological changes of the rats.ELISA method to detect the rat serum levels of GM-CSF and IL-4.Immunohistochemistry and RT-PCR to detecte the protein and gene expression of NF-?B in uterine tissues.(2)Reproductive group: After 20 days of administration,caged the rats of male to malein the reproductive group with a 7:2 ratio.To observe the emotional cycle of rats and the general situation of pregnant mice,recorded the litter size,average litter size,pregnancy rate and survival rate.Results 1 Anti-inflammatory experiments:After xylene-induced inflammation,compared with the blank group,the high,medium and low dose of The Gao's Prescription No.3 significantly inhibited the ear swelling caused by xylene in mice(P<0.05).2 Pharmacodynamics and reproductive experiments 1)Pharmacodynamics group(1)Behavioral changes in rats:The rats in the blank group behaved normally,after modeling,the rats showed lethargy of spirit,slow action and response,yellow and dull fur color,depilation,poor diet,increased vaginal secretions,and odor,and some of the later rats had poor defecation and abdominal distension.After drug treatment,the behavior gradually improved,and the rats in the high and medium dose groups of the Gao's Prescription No.3 recovere to normal.(2)Morphological changes of rat uterus tissue:(1)Visual inspection:In the model group,the uterus were congested and swollen,the lumens were narrow and deformed,a large amount of effusion or pus were found in the uterine cavity,poor elasticity,adhesion with surrounding tissues or heavy wrapping,some adhesion with the intestinal tube,intestinal obstruction,bilateral uterus are difficult to separate.After the treatment with the Gao's Prescription No.3,compared with the model group,the grading score of the anatomical morphology of the uterus in the drug administration groups were significantly decreased(P<0.05);(2)Rats uterine swelling:Compared with the blank group,the model group show significant uterine swelling.After treatment,the degree of uterine swelling in the high,medium and low dose groups were significantly reduced to compared with the model group(P<0.05).(3)Pathological morphology under light microscope:Model group:uterine tissue structure disorder,glandular disorder atrophy and deformation,visible infiltration of a large number of inflammatory cells,endometrial hyperemia and edema,degeneration and necrosis of epithelial cells,indicating the success of modeling.(4)Pathological score:Compared with the model group,the pathological scores of rat uterine tissue lesions were lower in the high,medium and low dose groups(P<0.05).(3)ELISA to detect the rat serum GM-CSF and IL-4 levels:The level of GM-CSF increased and the level of IL-4 decreased in rats in the model group(P<0.05).Compared with the model group,the GM-CSF levels of the rats in the high and medium dose groups were significantly reduced and the serum IL-4 levels were increased(P<0.05).Compared with the Pu Ling granule group and the Human placental tissue fluid group,the GM-CSF level of the rats in the high-dose group of the Gao's Prescription No.3 was significantly reduced and the IL-4 level was increased(P<0.05).(4)Immunohistochemical and RT-PCR to detect protein and gene expression of NF-?B in the rat uterus:The uterine NF-?B protein and gene expression in the model group were significantly increased(P<0.05).Compared with the model group,the uterine NF-?B protein and gene expression in the high,medium and low dose groups of the Gao's Prescription No.3 were significantly reduced(P<0.05);compared with the Pu Ling granule group,the NF-?B protein and gene expression of the high-dose group of the Gao's Prescription No.3 were significantly reduced(P<0.05).2)Reproductive group:(1)Changes in the estrus cycle of rats:The basic rule of the estrus cycle is observed in the blank group until pregnancy(4~5d).After modeling,some rats in each modeling group showed disturbance of estrous cycle and there were no statistically significant difference between each modeling group(P>0.05).During administration,the number of rats with estrous cycle disorder increased in the model group,comparing with the model group,the disorder rate of rats in the high-dose and medium-dose groups was improved(P<0.05).(2)General condition of pregnant mice:None of the rats in the model group are pregnant.After treatment with the the Gao's Prescription No.3,the pregnant rats had better mental state,more flexible activities,lustrous fur color,increased amount of drinking water,and faster increase of food intake and body mass.Compared with the blank group,there was no significant difference in the mean physical growth in the high-dose and medium-dose groups(P>0.05).(3)After the treatment of the Gao's Prescription No.3,the pregnant rate,pregnancy rate,litter number,litter size,average litter size and survival rate were increased significantly.Compared with the model group,the pregnancy rate and litter number of the high,medium and low groups were significantly increased(P<0.05),the average litter size of the high and medium dose group were increased(P<0.05).Conclusions 1 The Gao's Prescription No.3 can inhibit ear swelling degree caused by xylene in mice and has anti-inflammatory effect.2 The Gao's Prescription No.3 can effectively treat CPID.Comparison of efficacy,the high-dose group>medium-dose group>low-dose group and the efficacy of the medium-dose group is similar to that of the Human placental tissue fluid group,the low dose is equivalent to Pu Ling granules group.Its anti-inflammatory and antiadhesion mechanism may be related to the intervention of NF-?B signaling pathway.The Gao's Prescription No.3 up-regulates the level of anti-inflammatory factor IL-4,inhibits NF-?B activation and down-regulates the level of inflammatory factor GM-CSF.3 The Gao's prescription No.3 can improve the pregnancy rate of rats with CPID by controlling pelvic inflammation and inhibiting further damage of inflammation to fallopian tube and ovary.can improve the reproductive function of rats,inhibit further inflammation damage.The curative effect of each dose of The Gao's Prescription No.3 was better than that of the Pu Ling granules group,and the efficacy of the medium dose group was similar to that of the Human placental tissue fluid group.Figure 8;Table 17;Reference 105... |
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