Effect Of Dingkundan On CD146 And CD140b Gene Expression In Endometrial Tissue Of Rat Intrauterine Adhesion Mode

Objective: Through observing the effects of Dingkun Dan on the expression of EM SC-specific markers(CD146 、 CD140b)in the endometrium of IUA rats,and to investigate the possible mechanism of Dingkun Dan treating intrauterine adhesions.Methods: Sixty healthy(normal estrous cycle)SPF adult female SD rats were randomly divided into 6 groups with 10 rats in each group:sham operation group,model group,Dingkun Dan low,medium dose group,high dose group and estradiol valerate positive control group.Except for the sham operation group,the other groups of rats with intrauterine adhesions were established by mechanical combined infection double injury method.Sham operation group and model group were given normal saline.The low,medium and high dose groups of Dingkun Dan were given Dingkun Dan 1.10g/(Kg·d),2.20g/(Kg·d)and 4.40g/(Kg·d)respectively,and the estradiol valerate group was given progynova 0.06 g /(Kg·d).Each rat was intragastric once a day for 4 weeks,and after 4 weeks of continuous treatment we collected the uterus tissue.Analyzed the deposition degree of collagen fibers by Masson staining;observed the histomorphological changes of rat endometrium by HE staining;detected the expression levels of(CD146 、 CD140b)genes by Q-RT-PCR.Results: 1.Morphological observation: compared with the sham operation group,the endometrium of the model group was damaged and thinned,the glands were significantly reduced,the blood vessels were sparse,and the uterine cavit ies were partially dilated;compared with the model group,the morphology of uterine cavity in Dingkun Dan and estradiol treatment group recovered to varying degrees,the endometrium thickened,the number of glands and blood vessels increased.2.Masson staining: compared with the model group,the fibrosis in the model group was significantly higher(P<0.01),compared with model group,fiber deposition was significantly reduced in all drug treatment groups(P<0.01),compared with estradiol positive control group,the proportion of fibrosis area in Dingkun Dan low-medium dose group was higher(P<0.05),and the anti-fibrosis of Dingkun Dan high-dose group was similar to that of estradiol group(P>0.05).3.Detection of m RNA expression by Q-RT-PCR: compared with model group,Dingkun Dan and estradiol treatment groups could upregulate CD146 and CD140 b in uterine tissue of IUA rats,and the differences were statistically significant(P<0.05).CD140b: compared with Dingkun Dan low-dose group,CD140 b gene expression in medium-high dose group and estradiol group was significantly increased(P<0.05);compared with estradiol group,Dingkun Dan medium-high dose groups had no statistical significance(P>0.05).CD146: compared with Dingkun Dan low-dose group,the expression of CD146 gene in estradiol group showed an increasing trend but no statistical significance(P>0.05),while the expression of CD146 gene in medium-high dose group was significantly increased(P<0.05);compared with estradiol group,Dingkun Dan low,medium and high groups had no significant difference(P>0.05).Conclusion: 1.Dingkun Dan can effectively prevent and cure the intrauterine adhesions after double injury of endometrium caused by mechanical and infection in rats.2.The low expression of CD146 and CD140 b is related to the formation of IUA.3.Dingkun Dan may reverse fibrosis and repair damaged endometrium by up regulating the expression of EMSC(CD146,CD140b).4.Dingkun Dan dose up to the clinical equivalent dose(10.8g,Bid)or twice the clinical equivalent dose(21.6g,Bid)can better promote the expression of EMSC.