| Objectives Through experiment 1(pharmacodynamics experiment: to observe the behavior and uterine histomorphology of model rats,the changes of IL-2,FGF-2,PCT and hs-CRP in serum,and the protein and gene expression level of FGF-2 in uterine tissue)and experiment 2(analgesic experiment: to observe the analgesic effect of traditional Chinese medicine on writhing induced by glacial acetic acid in mice),Objective to comprehensively evaluate the intervention,efficacy and dose effect relationship of Gao's Pelvic Inflammatory Prescription No.5 on chronic pelvic inflammatory disease model rats and pain mice,and analyzed the anti-inflammatory,anti-adhesion and analgesic mechanism of the prescription.Methods In experiment 1,Twelve week old SD healthy female rats were divided into seven groups: blank group,normal saline group,Shaofu Zhuyu Granule group,high dose group,middle dose group,low dose group and human placenta tissue fluid group,10 rats in each group.In addition to blank group,the other group used phenol gel and mechanical damage method to establish CPID model.After the successful establishment of the 10 day model,all the 7 groups were treated for 20 days.In the course of the experiment,the behavioral changes of rats were observed,the pelvic condition and uterine pathological changes were observed under light microscope,and the levels of IL-2,FGF-2,PCT and hsCRP in serum and the protein and gene expression of FGF-2 in uterine tissue were detected.SPSS 25.0 was used to analyze the experimental data,and the difference was statistically significant(P<0.05).In experiment 2,60 Kunming mice were divided into model group,high dose group,middle dose group,low dose group,Shaofu Zhuyu granule group,placental tissue fluid group,10 mice in each group.All of them were given once a day for 7 days.One hour after the last administration,0.2ml of 0.6% acetic acid was injected into the abdominal cavity of each mouse.The writhing reaction was induced and the writhing times were observed.SPSS 25.0 was used to analyze the experimental data,and the difference was statistically significant(P<0.05).Results 1 Behavioral changes of rats: in the physiological saline group,the mental state was poor,the food intake was significantly reduced,the vaginal secretions were more,and the body weight was lighter than before.Rats in the blank group: the mental state was good and all aspects of physiological function were normal.In the course of treatment,compared with the normal saline group,the mental state of rats in each group gradually improved,the food intake increased,the vaginal secretion decreased,the body weight increased slowly.The rats in the high and middle dose groups of Gao's Pelvic Inflammatory Prescription No.5 were more significantly improved than those in other administration groups.2 Morphology of uterus of rats was observed with naked eye: in physiological saline group,the shape of uterus on the left side(affected side)was obviously enlarged,and there was a large amount of fluid or pus in the uterine cavity,which adhered to the surrounding tissue;the blank group,the high and middle dose groups of Gao's Pelvic Inflammatory Prescription No.5 were similar to the placental tissue fluid group: there was no significant difference between the left and right sides of the uterus,and there was no adhesion in the surrounding tissue.Gao's Pelvic Inflammatory Prescription No.5 low dose group and Shaofu Zhuyu granule group: the thickening and swelling of the left uterus(affected side)was slighter than that of the normal saline group,and some uterine cavity adhesion occlusion or a small amount of fluid accumulation could be seen,and slight adhesion with the surrounding tissue.3 The histomorphological changes of rat uterus were observed under light microscope:in the blank group,the delamination of uterine wall was clear,and no inflammatory cell infiltration was found.The high and middle dose groups of Gao's Pelvic Inflammatory Prescription No.5 and the placental tissue fluid group were similar to the blank group.Normal saline group: when the uterine cavity dilated,the uterine wall became thinner,when the uterine cavity proliferated obviously,the uterine wall thickened,fibroblasts proliferated,and a large number of inflammatory cells could be seen in the whole layer.Gao's Pelvic Inflammatory Prescription No.5 low dose and Shaofu Zhuyu granule group: the degree of uterine dilatation was slighter than that in the normal saline group,the morphology of epithelial cells showed proliferative degeneration and more inflammatory cell infiltration.4 The degree of uterine swelling in rats: compared with the normal saline group,the three dosage groups of Gao's Pelvic Inflammatory Prescription No.5 were significantly improved,and the differences were statistically significant(P<0.05).There was significant difference between the high and medium dose groups of Gao's Pelvic Inflammatory Prescription No.5 and Shaofu Zhuyu Granule group(P<0.05).5 The levels of IL-2,FGF-2,PCT and hs-CRP in serum of rats were detected by ELISA method: compared with the normal saline group,the level of IL-2 increased and the levels of FGF-2,PCT and hs-CRP decreased in the three dose groups of Gao's Pelvic Inflammatory Prescription No.5(P<0.05).Compared with Shaofu Zhuyu Granule group,the level of IL-2 increased and the levels of FGF-2,PCT and hs-CRP decreased in the high and medium of Gao's Pelvic Inflammatory Prescription No.5(P<0.05).6 Immunohistochemical method was used to detect the protein expression of FGF-2 in rat uterus: the protein expression level of FGF-2 in the three dose groups of Gao's Pelvic Inflammatory Prescription No.5 was significantly lower than that in the normal saline group(P<0.05).There was significant difference between the high and medium dose groups of Gao's Pelvic Inflammatory Prescription No.5 and Shaofu Zhuyu Granule group(P<0.05).7 The expression level of FGF-2 gene in rat uterus was detected by RT-PCR method:the expression level of FGF-2 gene in the three groups of Gao's Pelvic Inflammatory Prescription No.5 was significantly lower than that in the normal saline group(P<0.05).Compared with Shaofu Zhuyu Granule group,FGF-2 gene expression in high and medium dose groups of Gao's Pelvic Inflammatory Prescription No.5 was significantly lower(P<0.05).8 Results of analgesic experiment: compared with the model group,the dosage of Gao's Pelvic Inflammatory Prescription No.5 could reduce the writhing times of mice and improve the pain inhibition rate of mice(P<0.05).There was significant difference between the high and medium dose groups of Gao's Pelvic Inflammatory Prescription No.5 and Shaofu Zhuyu Granule group(P<0.05).Conclusions Gao's Pelvic Inflammatory Prescription No.5 has anti-inflammatory and anti adhesion effects,which can effectively treat chronic pelvic inflammatory disease in rats.1.It can improve inflammatory state,inhibit inflammatory exudation and promote inflammatory absorption,the mechanism may be related to down-regulating the levels of PCT and hs-CRP;2.Enhance the anti-inflammatory ability of the body,and its mechanism may be related to the up-regulation of IL-2 level.3.To reduce pelvic adhesion and prevent uterine fibrosis,the mechanism may be related to the down-regulation of FGF-2 level.4.It has analgesic effect,and the curative effect is related to the drug dose.Figure 4;Table 10;Reference 98. |
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