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Analysis of the Correlation of AMH and INS and Their Roles in Follicular Growth of PCOS Rats

Author: SunXiuHong
Tutor: WeiXiangCai
School: Jinan University
Course: Obstetrics and Gynaecology
Keywords: Animal models Polycystic ovary syndrome Anti- Mullerian hormone Insulin Insulin Resistance HOMA index Insulin receptor
CLC: R711.75
Type: Master's thesis
Year: 2011
Downloads: 138
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Abstract


Objective: To compare pregnancy hormone combined with human chorionic gonadotropin law DHEA law making SD juvenile female rats PCOS animal model differences, evaluation application results of the two methods, and to provide a suitable animal model for further study. Analysis of anti-Mullerian hormone, insulin and other influencing factors in PCOS rats follicle growth and development, change and the possible mechanism of action. 3. Explore PCOS serum anti-Mullerian and insulin resistance level correlation between ovarian partial and single follicle, and to provide new ideas for the clinical diagnosis and treatment of PCOS. 4. Analysis of the insulin receptor in the anti-Mullerian hormone and insulin resistance level of correlation in the role, to further explore the mechanism of interaction between the two. Methods: (1) the two methods produced PCOS rat model 120: the selection criteria farming juvenile female SD rats were randomly divided into two groups, law progestogen human chorionic gonadotropin (hCG) and dehydroepiandrosterone androsterone (DHEA) were produced rat the PCOS pathological model, through the vaginal smears, serum hormone levels, ovarian tissue pathological changes and to evaluate the effect of making the model. Detection and analysis of serum indicators: anatomy rats, abdominal aortic blood collected blood and serum was separated by radioimmunoassay (RIA) determination of serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH ); automatic chemiluminescent immunoassay analyzer detects serum estradiol (E2), progesterone (P) and the testosterone (T); ELISA method for the determination of serum anti-Mullerian hormone (AMH) and insulin (INS) value. Pathological examination of ovarian tissue: anatomical, separation, preservation groups in ovarian tissue fixed, paraffin-embedded biopsy conventional sections, HE staining, ordinary optical microscope to observe the pathological changes. 4 follicle count and impact factor analysis: ovarian tissue slices, each case of ovarian select five slices, each slice randomly selected five horizons, to count the number of follicles per field levels. With serum E2, T, LH, FSH, and LH / FSH values, AMH, FINS, FPG and HOMA-IR line correlation analysis, regression analysis underwent screening variables. Serum AMH levels and insulin resistance correlation analysis: calculated HOMA-IR, non-IR group (n = 10), according to the level of the rats were divided into PCOS PCOS merger IR group (n = 10), control group (n = 10), the line between the three groups of comparison and correlation analysis. Expression analysis of ovarian AMH and insulin receptor (Ir): Immunohistochemistry was used to detect the rats AMH expression and Ir, and image analysis to calculate the positive unit (PU value) line of semi-quantitative comparison and correlation analysis . Results: 1. Experimental group I (progestogen the hCG Act) and the experimental group Ⅱ (DHEA method) rats lost a proportion of regular estrous cycle were 86.4% and 77.3%, respectively; experimental group I showed obesity (P lt; 0.05) the experimental group, the experimental group Ⅱ weight significantly lower than the control group (P lt; 0.05); experimental group Ⅰ E2 compared with the control group (P lt; 0.05), the experimental group II control group (P lt; 0.05); II of LH was no difference in the control group; experimental group I, LH, LH / FSH ratio higher than that in the control group (P lt; 0.05). Two experimental serum T, FINS, FPG and HOMA-IR were higher (P lt; 0.05); rat ovarian pathology of the two experimental groups are visible the multiple expansion vesicles, granular cell layers reduce follicular film cells, stromal cell proliferation. 2. Experimental group Ⅰ middle, primary follicles, preantral follicles and the number of antral follicles compared with the control group increased significantly (P lt; 0.05), the number of primordial follicles, with no significant difference (P = 0.343) in the control group, the total follicle count is normal the control group, the number of follicles 2-3 times. The PCOS group number of follicles and serum testosterone and AMH positive correlation, Pearson correlation coefficients were 0.538 and 0.637 (P lt; 0.05). Multiple linear regression equation for the PCOS follicle count = 107.182 33.067T 0.412AMH. However, both the effect of serum AMH quite poor. Experimental group I serum AMH were higher (P lt; 0.05), a positive correlation (r = 0.691, P = 0.001); Multiple regression analysis shows T is removed, AMH and HOMA-IR, FINS between integrity line relationship (F = 12.362, P = 0.000), the equation: serum AMH = 15.776HOMA-IR 4.751 FINS-74.027, but the effect of both serum anti-AMH quite poor. Ovarian tissue immunohistochemistry of shows, AMH expression in preantral follicles and granulosa cells of antral follicles. The single follicle AMH levels among the three groups was no statistically significant difference (x2 = 0.657, P = 0.720). The number of follicles in the ovary AMH levels and AMH () significant differences in the presence of the three groups (F = 15.241, P = 0.000), the AMH expression intensity and AMH () The number of follicles was positively correlated. 5 ovarian tissue immunohistochemistry theca cells, granulosa cells and stromal cells, insulin receptor (Ir) are widely expressed in vascular endothelial cells express richer. Experimental group compared with the control group, PCOS no comparison between the IR and PCOS with IR group and the control group, Ir expression intensity There was no significant difference (P gt; 0.05); the local ovarian AMH and Ir expression intensity was no significant correlation (r = 0.000, P gt; 0.05); single follicle AMH Ir There was no significant difference (P gt; 0.05), no significant correlation (r = 0.011, P gt; 0.05). Conclusion: 1 progestogen the hCG method produced PCOS rat model is better than DHEA law, suitable for the study. May exist this role pathways: elevated insulin-stimulated androgen synthesis increased androgen cause an increase in the number of growing follicles in the ovary local, an increase in the growth of follicles synthetic excess of AMH AMH may inhibit recruitment of primordial follicles on the one hand, the other hand, lead to the stagnation of follicular development. PCOS rat single follicle AMH secretion is still normal, the experimental group increased AMH expression by AMH () to increase the number of small follicles result, the higher the degree of insulin resistance, the more small follicles, AMH and more. 4 of PCOS produce insulin resistance may occur after other aspects of the receptor function, or structural abnormalities and receptor, regardless of the number of receptors. The insulin receptor is involved in regulating PCOS ovarian interstitial fibrosis and angiogenesis involved in the occurrence and development of PCOS. 111

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CLC: > Medicine, health > Obstetrics and Gynaecology > Gynecology > Other diseases of the female genital > Ovarian disease
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