| Objective1 Through the study of the effect of Erbuzhuyu decoction which has removed psoralen on the expression of PROK1 mRNA in endometrium of patients with repeated IVF-ET failure,evaluate the clinical curative effect of subtraction supplemented soup decoction(decoction)and further explore its mechanism of action on endometrial receptivity.2 By investigating the effects of the Erbuzhuyu reduction prescription and it’s original prescriptions on the expression of endocytosis,ER,PR and LIF in the endometrium of blastocyst implantation dysfunction mice model,to explore the effects of Erbuzhuyu reduction prescription and it’s original prescriptions on endometrial receptivity of model mice and explore its pharmacodynamic mechanism.3 The experimental method of acute toxicology was used to administer the mice with the maximum dose and concentration of the two subtropical supplements within 24 hours.To observe whether the existence of acute toxicological effects of the Erbuzhuyu Decoction,provide more experimental evidence for the more extensive application of the old and famous Chinese medicine prescriptions and the safety study of the Erbuzhuyu Decoction.MethodClinical research:The patients with repeated IVF-ET failure were divided into two groups:the Erbuzhuyu reduction prescription and non-Chinese medicine group.Three months later,the endometrium in the mid-luteal phase of both groups was collected for biopsy,and the mRNA expression of PROK1 was measured by qCPR.With the endometrial biopsy method,a more realistic and objective exploration of the mechanism of action of the Erbuzhuyu reduction prescription.Experimental Study:Preparation of blastocyst implantation dysfunction mouse model with(GnRHa + HMG +HCG).1 Using electron microscopy scanning method to determine the morphological effects,and to contrast observation of the Erbuzhuyu Decoction,Erbuzhuyu reduction prescription,Bu Jiale,aspirin,and complementary Bu Jiale + aspirin The effect of each group of mouse endometrial pinopharyngeal expression.2 Use of immunohistochemistiy to study the molecular biological effects,to compare and test the Erbuzhuyu Decoction,Erbuzhuyu reduction prescription,Bu Jiale,aspirin and Bu Jiale plus aspirin,study and compare the effects of they have on ER,PR,LIF and integrin avB3 expression in endometrium of aspirin mice;comparison of traditional Chinese medicine monomer Rhizoma Drynariae and its constituents total flavonoids,naringin on endometrium ER,PR,LIF,integrin avβ3.3 The method of intragastric administration to the mice with a maximum concentration of suspension(without blocking the gastric tube)within 24 hours was used to determine if there was liver and nephrotoxicity in the Erbuzhuyu Decoction.Result1 In the clinical study,there was no significant difference in the expression of PROK1 mRNA in the endometrium between patients in the non-drug group and those in the traditional Chinese medicine intervention group(P>0.05),but the Chinese medicine intervention group was better in both the data dispersion and the mean of the data.2 In the scanning electron microscopy scan,observe the Erbuzhuyu DecoctionA(original prescription),Erbuzhuyu DecoctionB(remove psoralen),Erbuzhuyu DecoctionC(remove Rhizoma drynariae),Bu Jiale,aspirin,and Bu Jiale plus aspirin on the influence of the expression of the endocytosis of each group of mouse endometrium,it was found that the endometrium surface of the Chinese medicine group was more flat than that of the western medicine group.A large number of microvilli and prominent protruding pinopodes were found on the cell surface.In comparison with the three Chinese medicine groups,the interval between the pinopipothelial cells in the B group was more obvious,which was more conducive to cell adhesion.In the western medicine mice,the endometrium proliferative process of the endometrium was less developed,not full and different in morphology,and the cell boundary was less clear.3 Using immunohistochemistry to detect the expression of endometrial ER in the above-mentioned groups of model rats,the B group was better tan C group and A group model rats endometrium(P=0.0013,P=0.01).Compared with the western medicine group,the expression of ER in the endometrium of the aspirin + Bu Jiale group and Bu Jiale group micewas significantly higher than that of the A group(P = 0.002,P = 0.001);The expression of ER in the endometrium of the mice in Bu Jiale group was significantly higher than that in the C group(P=0.002).There was no significant difference between the B group and the western medicine group.However,the mean value of B group was significantly better than the aspirin group.In the expression of PR in mouse endometrium,the expression area of PR in endometrial uterus of mice in the B group was significantly higher than that in the A group and C group(P= 0.002,P=0.022);Compared with the western medicine group,the expression of PR in the endometrium of the mice in the C group was significantly lower than that of the aspirin + Bu Jiale group(P=0.022);There was no significant difference between the B group and the three groups of western medicine.However,the mean value of the B group was significantly better than that of the Bu Jiale group and the aspirin group.For the detection of LIF expression in mouse endometrium,the B groups was significantly better than A group(P=0.000).Compared with the Western medicine group,the expression of LIF in the endometrium,aspirin plus Bu Jiale group was significantly higher than that C group(P=0.000);The expression of αvβ3 in mouse endometrium was detected.Compared with the traditional Chinese medicine group,the area of expression in the B group was significantly higher than that in the C group(P=0.0025).On the average,the B group was better than in A group,and those in A group were better than those in C group;Compared with the wester medicine group,the expression of endometrial integrin avβ3 in the aspirin + Bu Jiale group mice was significantly higher than that in the C group(P=0.03);There was no significant difference between B group and A group and the western medicine three group,but the average value of the B group was significantly better than the western medicine three group.4 In the acute toxicology experiment,there were no obvious abnormalities in the vital signs,hair development,secretions,important organs,etc.in the mice of the control group and the experimental group.There was no change in body weight and liver and kidney function between the two groups of mice.No statistically significant difference(P>0.05).Conclusion1 Clinical studies have shown that Erbuzhuyu reduction prescription can upward adjustment expression of PROK1 mRNA in endometrium of patients with repeated IVF-ET failure,and thus promote the adherence of blastocysts to the endometrium,thereby improving endometrial receptivity.This provides a clinical basis for Chinese medicine to increase IVF-ET pregnancy rate.2 Experimental studies have confirmed that the Erbuzhuyu reduction prescription and it’s original prescriptions can improve the endometrial receptivity,the mechanism may be to increase the expression of the pinocytosis in the blastocyst implantation period,which is beneficial to blastocyst implantation,and the experiment also shows that Erbuzhuyu reduction prescription which has remove psoralen is the best.3 Experimental studies have shown that Erbuzhuyu reduction prescription and it’s original prescriptions can improve the expression of ER,PR,LIF and avβ3 in the endometrium of model mice.Therefore,the efficacy of the Erbuzhuyu reduction prescription and it’s original prescriptions is worthy of recognition.The mechanism of action of both may be through up-regulation of ER,PR,LIF,and avβ3 expression,thereby improving endometrial receptivity.4 Both Experiment 1 and Experiment 2 showed that the effect of Erbuzhuyu reduction prescription which has remove psoralen on improving the endometrial receptivity of model mice was more significant than that of the original prescriptions,and Erbuzhuyu reduction prescription which has remove Rhizoma drynariae was poorer than the former two groups.5 The mice administered with the maximum administration dose of the Erbuzhuyu Decoction did not exhibit acute poisoning.There was no significant change in appearance,body weight,morphology,and color of the main organs before and after the experiment and the normal group,and there was no significant difference in liver and kidney function between the two groups.The liver and kidney toxicity of psoralen monosomy was not demonstrated in the compound,and the Erbuzhuyu Decoction was safe and reliable in clinical application. |
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