| Objective Immunohistochemical staining was used to compare the expression of vascular endothelial growth factor(VEGFA)in endometrial adhesion patients and non-uterine adhesion patients.The significance of VEGFA expression in intrauterine adhesions was explored.Observe the expression of VEGFA in endometrial tissue of SD rats and the effect of Zuogui Pill on the expression of VEGFA in endometrial tissue of rats with IUA.Observe the effect of Zuogui Pill in improving the intrauterine of IUA rat model.The possible mechanism of membrane angiogenesis provides a theoretical basis for the clinical application of Zuogui Pill in the treatment of IUA patients.Methods In the first part of the clinical study,15 patients with mild to moderate intrauterine adhesions who were diagnosed with infertility,low menstruation,and amenorrhea were examined in our hospital by hysteroscopy.Fifteen patients without intrauterine adhesions underwent microsurgical treatment were the control group,and the postoperative results of the control group were proliferative endometrium.The corresponding endometrial tissue was collected during the examination and surgery of each group.The positive expression of VEGFA in endometrial tissue and normal endometrial tissue was detected by immunohistochemical method.The image processing software was used to measure VEGFA in high magnification field.Positive expression of the area of optical density(OD),the final statistical software analysis of each group of data.The second part of the experimental study,20 female SD rats were randomly divided into A and B groups,each with 10 rats.In the estrous model of rats,both groups of rats were treated with 100 °C hot water flow catheter to damage the left endometrium for 10 s.After the successful establishment of the model,group A rats were given intragastric gavage on the third day after the model was established.Rat Zuoguiwan suspension was given by gavage.Rats in each group were sacrificed 28 days later.Rats in each group were divided into left and right sides.The left side of group A was the model group and the right side was the control group.The left side of group B was the traditional Chinese medicine + model group.Right uterus for the traditional Chinese medicine group,a total of 4 groups.The uterine tissues of each group were divided into two parts,one paraffin-embedded section was fixed in 4% paraformaldehyde,HE staining was used to observe the endometrial tissue structure,cell morphology and glandular number in each group.The Masson staining was used to calculate the endometrial fibers.The areas were stained by immunohistochemical staining to observe the positive expression of VEGFA in each uterus tissue.The image analysis system was used to determine the ratio of the area of endometrial fibrosis in each group of rats and the optical density of VEGFA-positive expression area;the other was placed in liquid nitrogen.After 20 min,the cells were stored at-80 °C and the expression of VEGFA protein in the uterine tissues was detected by immunoblotting.Results The first part of clinical research:(1)VEGFA was expressed in both intrauterine adhesions and normal endometrium,and the positive expression products of VEGFA were brown-yellow particles in the cytoplasm of luminal epithelium and glandular epithelium.(2)The expression of VEGFA in the intrauterine adhesion group was significantly lower than that in the normal control group,and the difference was statistically significant(P<0.05).The second part of animal experiments:(1)Rats in each group were given 28 days after gavage.Morphology showed that the model group was narrow and atrophic in the middle part of the puncture,and the puncture wound was hyperplasia at the puncture wound.The proximal ovary end was obviously covered with fluid and adhered densely with the surrounding tissue.The group had no obvious hyperplasia and narrowness,and had no adhesion with surrounding tissues.(2)HE staining showed that part of the uterine cavity in the model group was stenosis or atresia,the level was fuzzy,the continuity of the luminal epithelium was interrupted,the matrix glands and blood vessels were missing;the uterine cavity of the model + Chinese medicine group was significantly larger than that of the model group,and the luminal epithelium was still intact Newborn glands and blood vessels can be seen in the stroma.Compared with the control group,the number of glands in the model group was significantly decreased(P<0.05).There was no significant difference in the number of glands in the Chinese herb group (P>0.05).Compared with the model group,the number of glands in the model + Chinese medicine group was significantly increased.(P<0.05).(3)Masson staining showed that a large number of blue-stained collagen fibers were observed in the uterine stroma of the model group,and a small amount of cord-like collagen fibers were seen in the stroma of the model + Chinese medicine group.No obvious fibrous adhesion was observed.The fibrosis area in each group was significantly higher than that in the control group(P<0.05).There was no significant difference in the Chinese medicine group(P>0.05).Compared with the model group,the model + Chinese medicine group was significantly reduced(P<0.05).(4)Immunohistochemical staining: Compared with the control group,there was no significant difference in the optical density of the VEGFA positive staining area in the uterine tissue of the traditional Chinese medicine group(P>0.05),and the positive expression of the model group was significantly reduced(P<0.05).Compared with the model group,the model + Chinese medicine group The positive expression was significantly increased(P<0.05).(5)Western blot analysis: Compared with the control group,the expression of VEGFA protein in each group had no significant difference(P>0.05),and the expression of VEGFA protein in the uterine tissue of the model group was significantly decreased(P<0.05);The expression of VEGFA protein in the model+TCM group was significantly increased(P<0.05).Conclusions(1)VEGFA was lowly expressed in endometrial tissue of patients with intrauterine adhesions,and low expression of VEGFA may be involved in the pathogenesis of intrauterine adhesions.(2)Zuogui Pill can effectively prevent and cure intrauterine adhesions in rats.Morphology and histology can be observed.Zuogui Pill can effectively restore the integrity of the intima of rats,improve the volume of uterine cavity,promote glandular and angiogenesis,and reduce internal Membrane fibrosis area.(3)Zuogui Pill can effectively prevent the occurrence and development of intrauterine adhesions in rats,and its mechanism may be achieved by up-regulating the expression of vascular endothelial growth factor VEGFA that promotes vascular endothelial regeneration in endometrial tissue. |
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