| Background and ObjectiveWith the continuous development of modern science and technology, peopleexperience fast pace of lif. Serious pollution of living environment to make theincidence rate of infertility had been increased year by year, which seriously affectedthe goodwill as between spouses and the harmonious development of the society. Theappearance of assisted reproductive technology (ART) provides a new solution forclinical treatment of infertility, and brings hopes to patients with infertility. As animportant treatment of ART, in vitro fertilization rate of In vitro fertilization andembryo transfer technique (IVF-ET) was90%, which is very high. However, theclinical pregnancy rate is still at a lower level (only30%to60%), and completelyunable to satisfy the needs of patients. There are many factors that influence thesuccess of pregnancy, including the thickness of endometrium, the quality of embryo,endometrial receptivity for embryonic and endometrial microenvironment. However,the endometrial thickness is the key factor for accept ability of endometrial embryosand growth of embryo. In recent years, the methods of increased endometrial thickness mainly include endometrial surgery with minimal damage and drug therapy.The endometrial surgery with minimal damage demands high level of doctors’operation technology and right time to trigger, so it’s not suitable for those doctorswho have less clinical experience. The drug therapy mainly includes growth hormone,estrogen, aspirin and Chinese traditional medicine prescription. Although all of themhave certain effects, they are not the best way. The price of growth hormone is tooexpensive for many patients. The security of estrogen and aspirin has many questions.The Chinese traditional medicine prescription is not convenient for taking, and patientcompliance is poor, so it cannot reach a better therapeutic effects. The Kuntai capsulewas a new kind of Chinese drug. It is very convenient to take. Clinical studies haveshown that it not only can improve the patient’s quality of ovarian, but also canincrease endometrial thickness of the infertility patients. But we could notcompletetly understand the mechanism of action of Kuntai capsule.The aim of the research was to observe the effections of Kuntai capsule onthickness of endometrium,the tissue morphology, expression of epidermal growthfactor (EGF), insulim-like growth factor-I (IGF-I), proliferrating cell nuclearantigenting(PCNA）and leukaemia inhibitory factor (LIF）in mouse after controlledovarian hyperstimulation and Super ovulation.Inaddition, we expiored its mechanism,experimental evident of Kuntai capsule is provided for seeking a new way ofincreasing clinical pregnancy rate.Materials and methodsWe selected clean unfertilized female healthy kunming mice as experimentalresearch object. The breeding condition of indoor humidity was60%~70%, indoortemperature was18℃~20℃,12hours of darkroom,12hours of light, free feedingand drinking water.We observed the estrus cycle of mice and choose continuousnormal rats oestrous cycle. Mice were randomly allocated into seven groups, andthere were fifteen mice in each group, including the control group (group A), thesuper ovulation group(group B), the super ovulation+lung low-dose of Kuntaicapsule group (filling Kuntai capsule90.1mg/100g weight,groupB1), the superovulation+large dose of kuntai capsule group [equal two times lung low-doses (groupB2)], the controlled ovarian hyperstimulation group [COH group (group C)], the COH+lung low-dose of Kuntai capsule group (group C1) and the COH+large dose ofkuntai capsule group [equal two times lung low-doses (group C2)]. The controlledovarian hyperstimulation model was built up according to the references. Each mousein the super ovulation group, the super ovulation+lung low-dose of Kuntai capsulegroup and the super ovulation+large dose of kuntai capsule group was intraperitonealinjected equal normal saline at9am every day, for nine days. PMSG10IU wasinjected on ninth day at9am for each mouse. While other mice in the controlledovarian hyperstimulation group, the COH+lung low-dose of Kuntai capsule groupand the COH+large dose of kuntai capsule group wsa intraperitoneal injected GnRHa1.5μg every100g weights at9am every day, for nine days. PMSG10IU was injectedon ninth day at9am for each mouse.The mice in the control group were killed in preoestrus, while the mice in thesuper ovulation group, the super ovulation+lung low-dose of Kuntai capsule group,the super ovulation+large dose of kuntai capsule group, the controlled ovarianhyperstimulation group, the COH+lung low-dose of Kuntai capsule group and theCOH+large dose of kuntai capsule group were killed after injecting of PMSG48hours later. The uterus was fixed and measured. Mouse endometrial morphology wasobserved with HE staining, mouse endometrial thickness was measured using imageanalyzer, and the expression of EGF, IGF-I, PCNA and LIF protein in endomentriumof mice were measured by immunohistochemistry sp techniques. All the data wereanalyzed using statistical software SPSS17.0. T-test and tˊ-test were used forquantitative data, and data was expressed mean±standard deviation.The inspectionlevel was0.05.Results1The endometrial thickness of mice in adding kuntai capsule group weresignificantly thicher than without kuntai capsule group （P＜0.01）, and theendometrial stroma of mouse were loose, the number of glands were more, thelumens were lager, they were uniform distribution.2The EGF and LIF protein mainly expressed in the cytoplasm of glandularepithelium cells, it expressed litter in interstitial cell cytoplasm. The IGF-1proteinmainly expressed in the cytoplasm of glandular epithelium cells, it expessed medium dose in interstitial cell cytoplasm. PCNA protein mainly expressed in endometrialcavity epithelium and glandular epithelium cells in the nucleus, it had a small amountof expression in interstitial cell nuclei.（P＜0.01）.3The expression levelsof EGF, IGF-I, PCNA and LIF protein in the superovulation group and COH group were significantly lower than the control group（P＜0.01）, and the expression levels of those four kinds of proteins in super ovulationgroup were significantly lower than the COH group（P＜0.01）.4The expression levels of EGF, IGF-I, PCNA and LIF protein in the superovulation group and COH group were significantly weaker than the add lunglow-dose of Kuntai capsule group and add large dose of kuntai capsule grouprespectively（P＜0.01）.5The expression levels of EGF, IGF-I, PCNA and LIF protein in add lunglow-dose of Kuntai capsule group were also significantly weaker than contrl groupand add large dose of kuntai capsule group respectively（P＜0.01）6The expression levels of EGF, IGF-I, PCNA and LIF protein in the controlgroup had no significantly different compared with the super ovulation+large dose ofkuntai capsule group and the COH+large dose of kuntai capsule group（P＞0.05）Conclusions1Kuntai capsule could significantly increase the endometrial thickness of miceand improve endometrial growth2Kuntai capsule could significantly increase the expression levels of EGF,IGF-I, PCNA and LIF protein in endometrial tissue of mice were treatment by thesuper ovulation and Controlled ovarian hyperstimulation.In addition, increasing thedose of Kuntai capsule could get better effection.3Kuntai capsule increased the thickness of endometrium maybe by increasingthe expression levels of EGF, IGF-I, PCNA and LIF protein in endometrial tissue ofmice were treatment by the super ovulation and Controlled ovarian hyperstimulation. |
