Kidney And Regulating Week The Aih Technology And Research

Objective1. To evaluate the treatment effect of kidney-reinforcing and menstrual cycle-regulation therapy in patients undergoing intrauterine insemination (IUI). To observe the effect of kidney-reinforcing and menstrual cycle-regulation therapy in follicle development and endometrial receptivity, and explore the mechanism of kidney-reinforcing and menstrual cycle-regulation therapy in improvement the clinical pregnancy rate.2. To research Yu Pao Decoction (a recipe of kidney-reinforcing and menstrual cycle-regulation) in rat's ovarian GC proliferation, secretion, cell cycle and the protein expression of EGFR,CX43and P38MAPK.To explore the mechanism of it in promoting follicular development from the level of the cell and molecular biology in order to provide scientific and objective experimental evidences to verify the effectiveness of the recipe.MethodClinical research1.60sterile women according with the clinical research criteria were randomlydivided into two groups. They were given the same clomiphene treatment.34cases in the control group were treated with clomiphene alone, and26cases in the study group were treated with kidney-reinforcing and menstrual cycle-regulation decoction in combination with clomiphene. The syndrome points of the deficiency of kindey qi of both groups after treatment were observed, and the relevance of the syndrome points of deficiency of kindey qi and endometrial thickness, pulsatility index (PI) and resistant index (RI) of endometrial blood stream, follicle blood flow and uterine artery were discussed.2. It was evaluated between both groups including the thickness and types of endometrium, pulsatility index (PI) and resistant index (RI) of endometrial blood stream, follicle blood flow and uterine artery and clinical pregnancy rate.Experimental study1. Preparation of water extraction and alcohol precipitation of Yu Pao Decoction, after filtered and stored at4℃。2. Determination was made the drug level of Yu Pao Decoction:We treated rat's ovarian GC with1×10-7g/ml～1×10-3g/ml of the decoction, and then respectively cultured for24h,48h,72h. At the same time, we set up a blank control group, through the MTT assaying different concentration of Yu Pao Decoction on GC growth and proliferation, determining the optimal dose-effort point to further experiments.3. Effect of Yu Pao Decoction on GC secretion, cell cycle, and the protein expression of EGFR, CX43and P38MAPK:GC was isolated from rat's ovaries and cultured for48 hours, divided into five group; three dosage groups of Yu Pao Decoction, blank control group and FSH(50ng/ml) control group at random.,after cultured for48h, assayed the secretion quantity of E2by chemiluminescence, used flow cytometry to assess the cell cycle change of GC, respectively detected the protein expression of EGFR,CX43and P38MAPK by Western-Blot method.Results1. The results of clinical research showed that after treatment the symptoms of deficiency of kidney qi in the study group were significantly relieveded comparing with the control group, the difference was significant (P<0.05).The clinical curative effect comparison of both indicated that the apparent rate and total effect rate in the study group were both higher than in the control group(P<0.05). The apparent rate of the study group was23.08%,the total effective rate was69.23%, while the apparent rate of the control group was11.76%, the total effective rate was41.18%; There were correlations between deficiency of kidney qi and endometrial thickness, pulsatility index (PI) and resistant index (RI) of endometrial blood stream, follicle blood flow and uterine artery. Negative correlation was displayed in deficiency of kidney qi and endometrial thickness, positive correlation in deficiency of kidney qi and pulsatility index (PI) and resistant index (RI) of endometrial blood stream perfusion, RI of follicle blood flow and uterine artery.2. Yu Pao Decoction could obviously promote the growth of endometrium and decrease the PI and RI of follicle blood flow, uterine artery and endometrial blood stream on the day of HCG injection in the IUI cycle. Compared with the control group, the difference was significant (P<0.05); Type A endometrium accounted for53.85%, B was38.46%in the study group on the day of HCG injection. In the control group,Type of A and B endometrium separately accounted for23.53%and29.41%. The study results also revealed that the clinical pregnancy rate and the cycle pregnancy rate of the study control were significantly higher than the control group (P<0.05). The clinical pregnancy rate and the cycle pregnancy rate of the study control were34.61%and19.56%. The control group was respectively11.76%and6.06%.3. Within the scope of1×10-6g/ml～1×10-4g/ml, the promoting proliferation capacity of Yu Pao Decoction gradually was enhanced increasing with the increase of the dosage and the culture-time. The best culture-time was48h.4. Each dosage group of Yu Pao Decoction could markedly increase secretion of E2compared with the blank control group (P<0.05), and the secretion quantity of E2gradually increased with the increase of the dosage. The difference was obvious between the low-dosage of Yu Pao Decoction and FSH control group (P<0.05). No significant difference of the secretion of E2was found between the medium,high-dosage group and FSH control group. 5. Though flow cytometry analysis operation, it showed that there was a remarkabe increase the in the ratio of GC Go/G1-phase and a obvious decrease in the ratio of S-phase and proliferation index compared with each dosage group of Yu Pao Decoction and FSH control group(P<0.05). The high-dosage group of Yu Pao Decoction could regulate up the ratio of S-phase and proliferation index. At the same time, regulate down the ratio of Go/G1-phase (P<0.05). There was no difference between the low,medium-dosage group of Yu Pao Decoction and FSH control group (P>0.05).6. EGFR protein expression gradually was increased with the increase of dosage of Yu Pao Decoction. The EGFR protein expression of each dosage group of Yu Pao Decoction was better than the blank control group (P<0.05).The high-dosage group of Yu Pao Decoction showed the most obvious effect (P<0.05), and the high-dosage group had the same effect with FSH control group(P>0.05). The EGFR protein expression of low-dosage group was significant lower than FSH control group (P<0.05).7. Three dosage groups of Yu Pao Decoction and FSH control group all could promote CX43expression. The CX43expression of low and medium-dosage group were lower than FSH control group (P<0.05). The high-dosage group could achieve the same effect with FSH group.8. Three dosage groups of Yu Pao Decoction and FSH control group all could promote P38MAPK expression. The difference was remarkable comparing with the blank control group (P<0.05). Compared with FHS control group, only decreased in low-dosage group, the P38MAPK expression in medium and high-dosage group had no difference (P>0.05).Conclusion1. Kidney-reinforcing and menstrual cycle-regulation therapy combined with clomiphene can significantly improve the clinical pregnancy rate of patients undergoing IUI.The possible mechanism of combined therapy is promoting oocyte maturation by benefiting follicle blood flow; enhancing endometrial receptivity by increasing endometrial thickness, changing the type of endometrium, reducing the blood flow resistance, increasing blood flow perfusion of uterine artery and endometrial blood stream.2. It is necessary to GC proliferation for follicle growth. Yu Pao Decoction can not only promote GC proliferation and secretion but also push cell cycle forward; as well as regulate up the CX43expression; enhance the ability of GC anti-apoptosis; adjust EGFR expression level; activate P38MAPK pathway,through above ways to achieve follicle growth and maturation.