The Influence Of Traditional Chinese Medicine (tcm) And The Receptivity Of Endometrium Experimental And Clinical Research

Objective1. On the basis of establishing the endometrial immunity microenvironment, this thesis investigates the effect of Zidan Decoction, on the bio-markers of endometrial receptivity and it discusses the molecular mechanism of endometrial receptivity is improved by Zidan Decoction.2. A self case-control study is carried out in this research in order to study the effect of Zidan Decoction on endometrial receptivity during the process of treating hypomenorrhea triggered by kidney deficiency or blood stasis.MethodsExperimental research3. Fresh endometrial tissues were obtained from pregnant mice on gd4.5days. Enzymolysis, filtration and pasted wall purification were adopted to isolate, purify and culture the endometrial stromal and glandular cells. With the primary culture technique, the vitro model of endometrial cells was set up and the purity of cells was identified by the method of immunohistochemistry.4. Mice’s Uterine Natural Killer(UNK) cells were obtained from pregnant mice on gd125days by the application of immunomagnetic microbeads and the cell purity was detected by a Flow Cytometer.The uNK cells were cultured in vitro and the supernatant of uNK cells were collected.5. Based on establishing of endometrial cells in vitro, mixed endometrial glandular cells and stromal cells at rate of1:1with uNK cells supernatants of different consistence from10%to90%and co-cultured them for24h and48h respectively. Chose an optimal concentration of uNK cells supernatants and set up uterine immune micro environmental system.6. Serum containing Zidan Decoction, Asprin and Blank Serum referrd to the Liyikui’s method were prepared with different concentrations of2.5%,5%,7.5%,10%and15%. The uterine immune micro environmental system was cultured in media with concentrations of serum containing the Zidan Decoction, Asprin and blank serum as the positive control, as well as2.5%FBS DMEM-F12culture media as the negative control’ for24h、48h、72h respectively. The cell proliferation effected by the Zidan Decoction containing serum in the uterine immune micro environmental system was detected by the MTT method so as to select the best consistence and time of drug action.7. Different concentrations of the Zidan Decoction were acted on the culture solution. FBS, blank and Aspirin control group were set up for contrast. The expression of LIF,integrin ανβ3and VEGF which were influenced by the Zidan Decoction in the immunity environment was assayed by Realtime-PCR.Clinical researchBy taking the method of self case-control study, the_menstrual volume was estimated by a pictorial chart of menstrual blood loss and the endometrial receptivity was assessed by using transvaginal color doppler sonography. The patients with hypomenorrhea due to kidney deficiency and blood stagnation, or those satisfied the inclusion criteria, were treated with the Zidan Decoction in three menstrual cycles. The effect of the Zidan Decoction was observed on improving endometrial receptivity of hypomenorrhea patients through menstrual quantity, Salle score, endometrial thickness, the inner shape of the endometrium, endometrial artery flow parameters and the endometrial blood flow states.Results1. Both the Zidan Decoction and Asprin containing serum could improve the expression of LIF, intergrin alpha(v)beta(3) and VEGF gene effectively of endometrial cells in the uterine immune micro environmental system(P<0.05).The effect of the Zidan Decoction containing serum is much better than that of Asprin, and there was no dose-effect relationship between them.2. The Zidan Decoction could increase patient’s menstrual quantity obviously (PBAC score, before treatment19.49±8.63, posttreatment48.19±13.86, P<0.001),promote the endometrial thickness(before treatment7.5±2.6mm, posttreatment9.1±1.8mm, P<0.001), perfect inner shape of the endometrium(before treatment A+B29/43; after A+B42/43),PI (before treatment PI2.85±2.22; posttreatment1.81±0.78, P<0.05) and the blood supply of endometrium locality (the endometrial blood flow only3cases existed before treatment but30cases posttreatment). Ultrasound average marks and sale score were increased (before treatment9.11±2.32; posttreatment16.14±2.70), and the endometrial receptivity was improved much better.Conclusion1. The uterine immune micro environmental system is preliminary established.2. The Zidan Decoction can improve the expression of LIF, intergrin alpha(v)beta(3) and VEGF gene effectively of endometrial cells in the uterine immune micro environmental system. Its efficacy is much better than Aspirin containing serum and there is no dose-effect relationship between them. Its affection on endometrial receptivity may be raised through promoting the expression of LIF, intergrin alpha(v)beta(3) and VEGF gene.3. The Zidan Decoction can increase patients menstrual quantity obviously, promote the endometrial thickness, perfect inner form of the endometrium, the supply of uterine artery blood and endometrium locality. Consequently, it can improve the endometrial receptivity effectively.