| Objective:Glucomannan gel is a new compound traditional Chinese medicine,which is made from extracts of Rhizoma baize,Centella asiatica and Fructus cnidii,and is being developed to treat gynecological inflammation.This topic on the basis of previous studies,the technology and composition of the improved drug the pharmacodynamics research,first of all,the first part through the establishment of the mouse ear swelling model to observe the glucomannan gel inhibiting effect on mouse ear swelling induced by xylene and ear tissue changes in concentrations of inflammatory factors in order to explore the anti-inflammatory effect;The second part explored the therapeutic effect of glucomannan gel on vaginitis by establishing a rat model of mixed infection vaginitis.In the third part,the therapeutic effect of glucomannan gel on cervicitis was explored by establishing a rat cervicitis model,and the mechanism of its anti-inflammatory effect was preliminally discussed whether the expression of MKP-1 was related.Methods:(1)The ear swelling model of mice was induced by xylene to observe the anti-inflammatory effect of glucomannan gel.7 days after administration,xylene was evenly applied to the inner and outer sides of the right ear,but not to the left ear,and the ear swelling rate caused by xylene was calculated.The contents of TNF-α,IL-6 and IL-1β in ear tissues of mice in each group were determined by enzyme-linked immunosorbent assay.(2)The rats model of mixed infection vaginitis was established by injecting staphylococcus aureus,Escherichia coli and Candida albicans into the vagina,and the therapeutic effects of glucomannan gel on mixed infection vaginitis were observed.After 7days of vaginal local administration,the contents of TNF-α,IL-1,IL-6,IL-8 and EGFR in serum of each group were determined by enzyme-linked immunosorbent assay.The vaginal mucosa tissues of rats were taken for pathologic examination.(3)To observe the therapeutic effect of glucomannan gel on cervicitis in rats,the model of cervicitis in rats was established by deep injection of phenol mucilage into vagina.After 7 days of vaginal local administration,the contents of TNF-α,IL-1,IL-6,IL-8and EGFR in serum of each group were determined by enzyme-linked immunosorbent assay.The cervical tissues of rats were taken for pathologic examination.The expression of MKP-1 mRNA in cervical tissues was detected by RT-PCR.Western blotting was used to detect the expression of MKP-1 protein in cervical tissues.Results:(1)Effect of glucomannan gel on ear swelling induced by xylene in mice:(1)Inhibition rate of ear swelling: the rate of ear swelling in model group was significantly increased(P<0.01).Compared with model group,ear swelling rate of mice in each administration group of glucomannan gel and compound metronidazole vaginal plug group was significantly reduced(P<0.01);There was no significant difference between gel matrix group and model group(P>0.05).(2)The contents of TNF-α,IL-6 and IL-1β in ear tissue of mice were detected by ELISA: compared with blank group,the contents of TNF-α,IL-6and IL-1β in ear tissue of mice in model group were significantly increased(P<0.01);Compared with model group,the contents of TNF-α,IL-6 and IL-1β in ear tissues of mice in each dose of glucomannan gel group and compound metronidazole vaginal plug group were decreased in different degrees(P<0.01 or P<0.05).(2)Effects of glucomannan gel on rat model of mixed infectious vaginitis:(1)Changes of TNF-α,IL-1β,IL-6 and IL-8 in serum detected by ENZYme-linked immunosorbent assay Compared with blank group,the contents of TNF-α,IL-1β,IL-6 and IL-8 in serum of model group were significantly increased(P<0.01).Compared with model group,the contents of TNF-α,IL-1β,IL-6 and IL-8 in serum of rats in each dose of glucomannan gel group and compound metronidazole vaginal plug group were significantly decreased(P<0.01),while there was no significant difference between gel matrix group and model group(P>0.05).(2)Changes of SERUM EGFR content in rats detected by ENZYme-linked immunosorbent assay: Compared with blank group,serum EGFR content in model group was significantly decreased(P<0.01);Compared with model group,the content of EGFR in serum of rats in each dose of glucomannan gel group and compound metronidazole vaginal plug group was significantly increased(P<0.01),but there was no significant difference between gel matrix group and model group(P>0.05).(3)Pathological morphologic examination of vaginal mucosa tissue: observed under light microscope(200x,40x),the model group had vaginal mucosa lamina corporeal edema,loose connective tissue arrangement,accompanied by more inflammatory cells scattered infiltration.Compared with the blank group,the inflammatory reaction was significantly aggravated.The vaginal squamous epithelium was covered in the administration group of glucomannan gel,and the epithelial cells were closely arranged and normal in shape.The lamina propria was rich in collagen and closely arranged,with very little inflammatory infiltration.(3)Effects of glucomannan gel on rat model of cervicitis induced by phenol mucilage:(1)Changes of TNF-α,IL-1β,IL-6 and IL-8 contents in serum of rats detected by ENZYme-linked immunosorbent assay: Compared with blank group,the contents of TNF-α,IL-1β,IL-6 and IL-8 in serum of model group were significantly increased(P<0.01);Compared with model group,the contents of TNF-α,IL-1β,IL-6 and IL-8 in serum of rats in different doses of glucomannan gel group and compound metronidazole vaginal plug group were decreased in different degrees(P<0.01 or P<0.05).There was no significant difference between gel matrix group and model group(P>0.05).(2)Changes of SERUM EGFR content in rats detected by ENZYme-linked immunosorbent assay: Compared with blank group,serum EGFR content in model group was significantly decreased(P<0.01);Compared with model group,EGFR content in serum of rats in different doses of glucomannan gel group and compound metronidazole vaginal plug group was increased in different degrees(P<0.01 or P<0.05).There was no significant difference between gel matrix group and model group(P>0.05).(3)The cervical tissue pathological morphology:observed under light microscope(200 times and 40 times),cervical squamous epithelium covering model group,a few scattered in neutrophils infiltrating,visible cortex on the lamina propria rich collagen,closely packed,visible more inflammatory cells are scattered in the infiltration,including neutrophils,lymphocytes,and so on,compared with the blank group,mild increase inflammation reaction.The cervical covering squamous epithelium of the glucomannan gel administration group was normal in shape,with abundant collagen and compact arrangement of lamina propria,and a few inflammatory cells scattered and infiltrated.(4)Results of RT-PCR and Western blotting:(1)The expression of MKP-1 mRNA in cervical tissue of model group was significantly decreased compared with that of blank group(P<0.01);Compared with the model group,the mRNA expression of MKP-1 in the cervical tissues of rats treated with glucomannan gel was increased in different degrees(P<0.01 or P<0.05).(2)The expression of MKP-1 protein in cervical tissues was detected by Western blotting: Compared with the blank group,the expression of MKP-1 protein in cervical tissues of model group was significantly decreased(P<0.01);Compared with the model group,the expression of MKP-1 protein in the cervical tissues of rats treated with glucomannan gel was increased in different degrees(P<0.01 or P<0.05).Conclusion:1.By establishing a mouse ear swelling model,the mice were induced to produce acute inflammatory state.The comparison of the degree of ear swelling and the content of inflammatory factors in the ear tissue between the drug administration group and the model group showed that the glucomannan gel had good anti-inflammatory effect.2.By establishing a rat model of mixed infectious vaginitis,the changes of inflammatory factors and EGFR in serum and the pathological changes of vaginal mucosa were detected,indicating that glucomannan gel has a good therapeutic effect on vaginitis.3.By establishing a model of cervicitis in rats,the changes of serum inflammatory factors,EGFR content,histopathological changes of cervical tissues and expressions of MKP-1 mRNA and protein in cervical tissues were detected,indicating that glucomannan gel has a good therapeutic effect on cervicitis.The improvement of inflammatory response may be related to the promotion of MKP-1 expression. |
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