| Objective:To explore the effect of Huluan decoction on the thickness and morphology of endometrium in rat model of thin endometrium.To reveal the mechanism of promoting the proliferation of thin endometrium by detecting the expression of mRNA and protein in endome trium.To provide theoretical basis for clinical treatment and scientific research.Methods:60 SD rats were randomly divided into six groups:blank group,model group,aspirin group,low,medium and highHuluan decoction groups.10 rats in each group.The other five groups,except the blank group,made a thin endometrial model by intrauterine perfusion of anhydrous ethanol for 5 minutes.The rats in the model group and the blank group were given gastric perfusion with distilled water,and the other grous were given to the intragastric administration respectively by Aspirin,low,medium and highHuluan decoction.Each group was perfused continuously for 7 days.After three estrous cycles,The thickness of endometrium was processed by leica Qwin Plus image.and the morphology of endometrium was observed by HE staining;The endometrial tissue in blank group,model group,aspirin group and medium dose Huluan decoction group were selected to detect the expression of VEGF、VEGFR-2、PI3K and Akt mRNA by real-time PCR,and the expression of VEGF、VEGFR-2、PI3K and Akt protein in endometrial tissue by western-blotting.Results:1.The thickness of endometrium(cm)in blank group was 0.89±0.17,in model group was 0.10±0.04,in aspirin group was 0.65±0.22,in low dose group was 0.33±0.10,in middle dose group was 0.56 ±0.07,in high dose group was 0.71±0.11.The thickness of endometrium in model group was significantly thinner than that in blank group.The difference was statistically significant(P<0.05);compared with the model group,the endometrium of aspirin,low,medium and highHuluan decoction group were thickened to varying degrees,and the difference was statistically significant(P<0.05);compared with the aspirin group,the low dose Huluan decoction group were significantly increased(P<0.05).the endometrium of low dose Huluan decoctiongroup group were thinner,and the difference was statistically significant(P<0.05);there was no significant change in intimal thickness in the high、middle dose group(P>0.05).The comparison of endometrial thickness among the three dosage groups showed that:high dose group>middle dose group>low dose group,and the difference was statistically significant(P<0.05).2.Endometrial morphology:in blank group:the number of endometriaj glands was more,the glands were well developed,the glandular cavity was large,the interstitial cavity was loose,the glandular epithelium was high columnar,and the interstitial vessels were obviously proliferating.Compared with the blank group,the endometrium was thinner and the glands were dysplasia.Compared with the model group,aspirin group and Huluan decoction group could improve the morphology of endometrial tissue.The thickness of endometrium and the number of glands in the model group were significantly higher than those in the model group.The gland developed well,the glandular cavity was large,the interstitial cavity was loose,the glandular epithelium was high columnar,and the interstitial vessels were obviously proliferating.3.The results of real-time PCR showed that the expression of VEGFR VEGFR-2、PI3K and Akt mRNA in model group was the lowest in the four groups,and the expression of VEGF、VEGFR-2、PI3K and Akt in Huluan decoction group and aspirin group was significantly higher than that in model group.The difference was statistically significant(P<0.05),but there was no significant difference between aspirin group and Huluan decoction group(P>0.05).4.Western-blotting results showed that the protein expression of VEGF and VEGFR-2 in the model group was the lowest in the four groups,and the protein expression in the Chinese medicine group and aspirin group was significantly higher than that in the model group.The difference was statistically significant(P<0.05),but there was no significant difference between aspirin group and Huluan decoction group(P>0.05).The expression trend of PI3K and Akt protein was similar to that of VEGF and VEGFR-2-Conclusion:1.Huluan decoction can improve the morphology of endometrium and promote the hyperplasia of endometrium.It has obvious therapeutic effect,and the higher the dosage is,the stronger the effect is.2.The expression of VEGF、VEGFR-2、PI3K and Akt in endometrium of rats was up-regulated by Huluan decoction and aspirin.VEGF and VEGFR-2 were closely related to vascular regeneration and tissue repair,and its high expression could promote the repair of thin endometrium.It suggested that Huluan decoction could activate the PI3K/Akt signaling pathway by up-regulating the expression of VEGF and VEGFR-2 to induce the repair of the thin endometrium in rats. |
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