Changes in E-Cadherin Expression Levels between Obese and Normal Rat after Gonadotropin Hormone Stimulation

Siti Nurbaya¹, Numlil Khaira Rusdi²*, Diyah Kristianti¹, Eldafira³, Nurhuda Sahar³,

Rosmalena⁴, Conny Riana Tjampakasari¹, Berna Elya⁵, Rozana Othman⁶, Maifitrianti⁷,

Anna Wirdiani Fathiah⁸

¹Department of Pathology Clinic, Faculty of Medicine, University of Indonesia.

²Magister of Pharmacy Education Program, School of Postgraduate,

Universitas Muhammadiyah Prof. Dr. Hamka, Jakarta, Indonesia.

³Department Biology, Faculty of Medicine, University of Indonesia, Jakarta.

⁴Department of Chemistry, Faculty of Medicine, University of Indonesia, Jakarta.

⁴Department of Microbiology Faculty of Medicine, University of Indonesia, Jakarta.

⁵Faculty of Pharmacy, University of Indonesia, Jakarta.

⁶Faculty of Chemistry, University of Malaya.

⁷Faculty of Pharmacy, Universitas Muhammadiyah Prof. Dr. Hamka, Jakarta, Indonesia.

⁸Master Program in Biomedical Science, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia.

*Corresponding Author E-mail: numlilkhaira@uhamka.ac.id

ABSTRACT:

Overweight (Obesity) and administration of ovarian stimulation regimen have a negative impact on endometrial receptivity during implantation. It is known that the receptive condition of the endometrium during the embryo implantation reception period is characterized by the expression of various receptivity markers, one of which is E-Cadherin. The purpose of this study was to analyze and compare the level of E-Cadherin expression in the endometrium of normal weight female rats with a group of stimulated normal weight rats, a group of overweight rats and a group of stimulated overweight rats. overweight, obesity and normal weight in the stimulated and unstimulated groups. Methods, 3-month-old female Wistar rats with a normal body weight of around 200grams and overweight rats (given a high-calorie fat diet) for 2-3months), an average body weight of 275grams. Normal weight rats were stimulated once with recombinant FSH (Gonal F) intraperitoneally at a dose of 15–20IU and overweight rats were injected with a dose of 20-25 IU FSH in the diestrus phase. 48 hours after injection (estrus phase), the rats were mated with male rats (Vaginal plug). Endometrium sampling was performed serially on days 3, 4, and 5 after mating. E-Cadherin expression was examined using immunohistochemical staining (IHK). Using the IHK score calculation formula = (% positive high cell contribution x 4) + (% positive cell contribution x 3) + (% positive low cell contribution x 2) + (% negative cell contribution 1), Results, E-Cadherin expression in the endometrium of rats in the normal weight group did not show a significant difference in expression levels compared to the stimulated normal weight rat group (P = 0.07). When compared with the group of overweight mice that were not stimulated, there was a significant increase in expression levels (P = 0.02). Furthermore, with the expression of E-Cadherin in the group of overweight mice that were stimulated, there was a significant increase (P = 0.01). Conclusion, E-Cadherin expression increased significantly in overweight female mice, both stimulated and unstimulated, compared with normal weight mice.

KEYWORDS: Obesity, E-Cadherin, Recombinant-FSH, Endometrium, Immunohistochemistry.

INTRODUCTION:

Obesity cases in women continue to increase every year, even reaching alarming levels¹. Women aged 12-19 year experience an increase in obesity of up to 20.6% and tend to be obese at age 35 compared to women with normal weight^2,3. According to longitudinal studies, 80% of children with severe obesity and 56% of children with obesity grow into obese class II (BMI 35–39.9kg/m2) and III (BMI 40–49.9kg/m2) adults⁴. The National Center for Health Statistics (NCHS) recently reported that women are significantly more likely to be obese (11.5%) than men¹. The increasing number of obese women at reproductive age requires a review of its impact on women's reproductive success, both naturally and with assisted reproductive techniques^5,6,7.

The impact of obesity on patients undergoing treatment through assisted reproductive technology shows poor implantation and pregnancy rates compared to women with normal weight^8,9,10. In women (BMI > 40kg/m2) there was a decrease in the live birth rate due to the high miscarriage rate (22.2%) in women who became pregnant through IVF/ICSI technology, compared to women with normal weight, which was 12.6%. in women with normal weight¹¹. Other studies also revealed similar findings, with significant odds ratios of 0.984(95% CI 0.972-0.997) and 0.981 (95% CI 0.972-0.997) and 0.981(95% CI 0.967-0.995) indicating a progressive decrease in pregnancy and live birth rates per unit BMI kg/m2^12,13. In addition, the cumulative pregnancy rate after four IVF cycles decreased with increasing BMI^12,14,15. However, the results of other studies showed little/no effect of BMI on oocyte quality, endometrial receptivity, and pregnancy outcomes¹⁶. The underlying mechanism of the poor outcome of increased BMI, particularly with regard to the endometrium, remains to be fully elucidated^17,18. It appears that the decreased pregnancy rate in obese patients undergoing IVF may be associated with impaired endometrial receptivity during the implantation period¹⁹.

E-cadherin is a transmembrane protein that functions primarily to mediate cell-to-cell adhesion through adherens junctions. Alterations in the expression of endometrial receptivity markers such as HOXA10, Integrins and leukemia inhibitory factor (LIF) involved in endometrial receptivity have been reported in infertile women.

Related to the lower implantation rate in obese women compared to normal women in infertile patients undergoing IVF and no studies have assessed the difference in E-Cadherin expression as a marker of endometrial receptivity during the implantation window in obese women and obese women undergoing in vitro infertility.

It is suspected that there is a significant correlation between the overweight factor and the impact of the stimulator regimen on endometrial receptivity during the implantation period. In this study, experimental animals were used, considering that it is unethical for women participating in IVF. The aim of this study was to analyze and compare the expression levels of endometrial receptivity markers (E-Cadherin) in normal weight female Wistar rats compared to normal weight rats that were stimulated, with overweight rats that were stimulated and without stimulation.

MATERIALS AND METHODS:

Animal experiment:

The use of female Wistar rats as experimental animals was approved by the ethics committee of the Faculty of Medicine, University of Indonesia (ethics number: 1105/UN2.F1/ETIK/PPM.00.02/2024). Instructions that were followed when working with the animals were part of the guidelines for the use and care of experimental animals. For a period of two weeks, 36 adults female Wistar albino rats, ages around 3.5 months, weighing between 150 and 200grams for normal weight and 250 to 300grams for obese rats, were acclimated to a 12-hour light and 12hour dark cycle. Overweight rats were obtained through the procedure of giving 20grams of high fat and high sugar diet every day for 8 weeks and weighing is done every week. Calculation of excess body index using the Lee index formula, with the criteria of index value >0.3. In this investigation, only female rats with regular cycles were employed; those without a normal estrous cycle were removed. Each of 9 female rats from the normal weight and obese rat groups were divided into two groups: the control group and the group receiving recombinant FSH stimulation²⁰. Rats with normal weight received a dose of 20 IU FSH, while the obese rat group received a dose of 25 IU. FSH injection was performed intraperitoneally (ip) at the beginning of the diestrus phase^20,21,22. Two days after FSH administration, female rats were mated with male rats, (successful mating was indicated by a vaginal plug). Uteruses were taken serially on days 3, 4 and 5 after mating. Uteruses were stored in BNF (normal buffer formalin) until immunohistochemistry (IHC) measurements were performed.

Immunohistochemistry for E-Cadherin:

0.1mL/kg BW of ketamine was used to anesthetize the animals prior to surgery. A third of the uterine organs, one on each side, were dissected and preserved in a 10% solution of normal buffer formalin (BNF). then embedded in paraffin after being dehydrated in serial-grade alcohol and xylene solution. Hematoxylin-eosin (H-E) staining was applied to paraffin-embedded samples that were sectioned at a thickness of 4μm for histological dating if required. The endometrial tissue that was fixed in paraffin blocks was sliced thinly, ranging in thickness from 0.3 to 0.5μm. The glass object was covered with APEX.

The procedure was dehydrated in an alcoholic solution following paraffinization in a xylol solution. A 0.05M PBS solution with a pH of 7.2 was used to clean the slides. The slides were incubated in an H2O2 solution for 10 to 15 minutes before being washed with water. Following their immersion in the retrieval buffer solution, the slides were heated for 30minutes in RG1. The Rabbit anti-E-cadherin monoclonal antibody GT 477 Gene Tex was diluted 1:500 on cooled slides after they had been cleaned under running water.

The slides underwent a 60-minute incubation period at room temperature. To clean the slides, a buffer in solution was utilized. The slides were washed again in PBS solution after being incubated for 25minutes at room temperature after Polymer HRP was added. Five minutes passed after we added one drop of color. To keep the slides clean, water was flowing. The slides were washed under running water after HE staining for one minute. The slides were sealed with mounting medium and examined under a light microscope.

To compare the results of the samples, normal endometrial tissue and Ca mammae tissue were used as positive and negative controls. Slides were seen at 400x magnification under illumination. The five area images were selected at random, and the intensity of the brown color and the cell counts were computed using ImageJ analysis software.

Cells expressing E- Cadherin were counted in the rectoanal epithelium using an IHC profiler. Next, rectoanal epithelial cells expressing E-Cadherin were counted from 3 representative fields of view with 400x magnification (carried out by 2 technicians, blind reading). Calculation uses algebraic formula^23,24. Calculation method: IHC score = (% contribution of high positive cells x 4) + (% contribution of positive cells x 3) + (% contribution of low positive cells x 2) + (% contribution of negative cells x 1)

Statistical Analysis:

The maximum-minimum and median values were used to describe the data. The SPSS 22 program was used to compare the immunoreactive scores of the stimulated group and the normal control group and determine whether the differences were significant. Assessing the data's homogeneity and normality was the initial stage in this investigation.

One-way ANOVA was used as the statistical test if the distribution was homogeneous and normal. If the ANOVA analysis revealed significant differences, the Tukey HSD (Honestly Significant Difference) test was used to determine how one group differed from another. The Pearson correlation was employed in correlation analysis to determine whether the data was regularly distributed. The Spearman correlation analysis was used in cases where the data was not regularly distributed. The significance limit for the statistical test decision is set at 5% (p = 0.05).

RESULT:

The analysis involved 36 endometrial tissues from female WISTAR strain Rat that were taken during the implantation period (i.e. days 4, 4 and 5 after mating with male Rat. The endometrial tissue was obtained from four different groups of rats: group 1 was normal and unstimulated, group 2 was normal weight and stimulated with 15 IU of recombinant FSH, group 3 was obese and not treated, and group 4 was obese and stimulated with 20 IU of recombinant FSH. A total of 30 endometrial samples that were successfully stained immunohistochemically obtained varying levels of E-Cadherin expression, ranging from weak to strong expression levels, as shown in Figure 1. A-D. The average intensity of dark brown color appeared in obese mice and was moderate in normal unstimulated mice and low in normal stimulated mice (Figure 1A-D).

Immunohistochemical staining was used to determine the cellular distribution and E-cadherin quantification. The expression was found in the luminal gland epithelial cytoplasm. Every one of the 36 samples that were analyzed tested positive for immunohistochemical staining, which revealed a deep brown color. The intensity that shows up ranges from modest to very intense. Figure 2 shows that the expression of E-cadherin is higher in obese rats than in rats of normal weight. Compared to the normal rat group, the fat rat group's brown colouring was more pronounced.

Immunocytochemical (IHC) analysis of the endometrium of rat at the implantation stage, normal and overweight rat groups showed uneven distribution in the endometrium (Figure 2). E-Cadherin expression was highly brown in the endometrial luminal epithelial cells and functional stromal cells of unstimulated overweight rats (Figure 2A), and it was lower in the endometrium of overweight rats stimulated with FSH than in the unstimulated group in both the luminal epithelium and functional stromal layers (Figure 2B). Furthermore, the expression level of E-Cadherin in the luminal epithelial cells and glandular epithelial cells of the endometrium was rather strong (Figure 2C). In contrast, the degree of E-Cadherin expression in the luminal epithelial cells and functional stromal cells of the endometrium was weak in the normal rat group without stimulation (Figure 2D).

Table 1 shows the expression levels of E-Cadherin studied in 4 groups of mice. The average expression levels of E-Cadherin in female mice in the normal weight group did not show any significant difference compared to the group of normal weight mice that were stimulated. Furthermore, when compared to the group of overweight mice without stimulation, there was a significant increase in expression levels (p = 0.02), as well as in the group of overweight mice, there was a significant increase in export levels (p = 0.01).

Figure 1. Expression of E-cadherin upon IHC staining. Intensity of positive staining in the cytoplasm of the luminal epithelium and endometrial stromal cells. (A), unstimulated obese rat (B) stimulated obese rat (C), unstimulated normal rat (D), stimulated normal rat E), Positive control of mammary ca; (F) Negative control

Figure 2. Comparison of E-cadherin expression in normal rat and obese rat.

DISCUSSION:

The glycoprotein E-cadherin, an epithelial cadherin and one of the classic cadherins, is present on the surface of cells and oversees implantation and embryonic structural alterations. Luminal and glandular epithelium express the adhesion molecule cadherin. The maintenance of intercellular connections by E-cadherin is dependent on calcium²⁵. Whether it plays a role in the implantation of embryos is still uncertain. On the third day of pregnancy, a single uterine horn was injected with varying concentrations of E-cadherin antibody in each animal used in the intrauterine horn injection model. The rat given three micrograms of E-cadherin antibody had considerably less embryo implantation, according to the data²⁶.

By interacting with catenin, cadherin can attach itself to the actin cytoskeleton. Another crucial aspect of regulation is the relationship between α-catenin and the actin cytoskeleton¹⁹. ZO-1, vinculin, and α-actinin are among the actin-binding proteins that α-Catenin interacts with. According to kinase activation, adhesion is also regulated by tyrosine phosphorylation of the cadherin-catenin complex. Another protein that regulates adhesion activity is p120ctn, which shares structural similarities with β-catenin, a protein that contains armadillo repeats²⁷. In contrast to the traditional catenin binding site, it attaches to the juxta membrane domain. The cadherin-mediated adhesion process also involves the small GTPases Rac, Rho, and Cdc42²⁶.

The immunocytochemistry method was used to examine the expression of E-cadherin in female rats' endometrium during the implantation phase. It was shown to be substantially expressed in the luminal epithelium and moderately expressed in the glandular epithelium²⁸. This finding seems to indicate the adhesive properties of E-Cadherin as an adhesive protein of the embryo with the endometrial epithelium during the implantation process (Figure 1). E-cadherin expression has been shown to rise dramatically in the apical membrane of rat uterine epithelial cells during preimplantation stage²⁹. In order to adhere the embryo to the endometrial epithelium during the implantation phase, E-Cadherin plays a critical role in the uterine luminal epithelium³⁰.

Endometrial stimulation methods have been shown in numerous studies to be detrimental to the success of IVF treatments^31,32. Further studies have also shown the negative effects of obesity and the benefits of weight loss for IVF and conception. A sign of successful implantation is the pattern of elevated E-cadherin expression during the peri-implantation phase in women with normal body weight. Pregnancy rates rise when weight loss is combined with diet and exercise, according to meta-analysis research by Best et al, 2017³⁰. Thus, the pattern observed in rats throughout the implantation stage corresponds to the rat uterus's E-cadherin expression profile³³.

In certain instances, particularly in endometrial cancer, obese women's endometrium has elevated expression of E-Cadherin. This is positively correlated with certain hormones. Other hands, the mechanism of changes in E-cadherin expression in female infertility is still largely unknown. During the implantation window period in the mid-secretory phase, endometrial receptivity depends on molecular processes regulated by hormones. Progesterone resistance may also affect the expression of E-Cadherin molecules³⁴.

The data obtained are different from the initial hypothesis, where body weight and stimulation factors will affect E-Cadherin expression, thus negatively affecting endometrial receptivity. Nevertheless, other research findings indicate that E-cadherin expression in glandular and luminal epithelial cells during the mid-secretory phase in healthy fertile controls is either very low or nonexistent. However, in the mid-secretory phase, it was shown that the luminal and glandular epithelial cells of infertile individuals expressed significantly more E-cadherin than did healthy, fertile controls (B). These findings, which are in line with the findings of other previous animal investigations, imply that E-cadherin protein expression may need to be temporally downregulated or deleted during the implantation window in order to allow for blastocyst invasion and epithelial cell separation^35,36. Our study on experimental animals obtained similar results, where there was an increase in the level of E-Cadherin which is significant in glandular and luminal epithelial cells during the implantation period³⁷. Although the negative impact of high expression during the implantation window on implantation success has not been explained, when connected with our data on the number of children born (data unpublish) a negative correlation was obtained.

E-cadherin expression and dispersion must occur in a timely manner for normal compaction and blastulation. It has been discovered that the protein's redistribution to regions of cell-to-cell contact is either nonexistent or irregular. The common inability of aberrant embryos to compress (and later blastulate) normally may be directly related to their inability to correctly relocalize E-cadherin.

E-cadherin expression is epigenetically controlled during trophoblast invasion, as our lab has recently demonstrated³⁸. It has been discovered that there is either no redistribution of the protein to regions of cell-to-cell contacts or that it occurs irregularly. One possible explanation for the frequent inability of aberrant embryos to compress (and later blastulate) normally is their inability to correctly relocalize E-cadherin. Lack of physical contact between blastomeres or developmental asynchrony may be the cause of E-cadherin failure to move³⁹. The limitation of this research is the less-than-optimal success of the fattening process for female rat. During a month and a half of the fattening process, the average weight of the rat did not exceed 300 grams. In this research, it is better to use the term overweight.

CONCLUSION:

E-Cadherin expression increased significantly in overweight female mice, both stimulated and unstimulated, compared with normal weight mice.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

ACKNOWLEDGMENTS:

This research was funded by PUTI Q2, 2023, a grant from Indonesia University. Contact number NKB-247/UN2.RST/HKP.05.00/2023

REFERENCES:

1. Hales CM, Carroll MD, Fryar CD, Ogden CL. Prevalence of obesity and severe obesity among adults: United States. 2017-2018. NCHS Data Brief. 2020; 36(1): 1–8.

2. Hales CM, Carroll MD, Fryar CD, Ogden CL. Prevalence of obesity among adults and youth: United States, 2015-2016. NCHS Data Brief. 2017; (288): 1–8.

3. Woo JG, Zhang N, Fenchel M, Jacobs DR, Jr, Hu T, et al. Prediction of adult class II/III obesity from childhood BMI: The i3C consortium. Int J Obes. 2020; 44(5): 1164–1172.

4. Sermondade N, Huberlant S, Bourhis-Lefebvre V, Arbo E, Gallot V, Colombani M, Fréour T. Female obesity is negatively associated with live birth rate following IVF: a systematic review and meta-analysis. Hum Reprod Update. 2019; 25(4): 439–451.

5. ASRM. Practice Committee of the American Society for Reproductive Medicine Obesity and reproduction: a committee opinion. Fertil Steril. 2015; 104: 1116–1126.

6. Oliveira JB. Obesity and Reproduction. JBRA Assist Reprod. 2016; 20: 194–259.

7. Mahadeva Rao U.S., Thant Zin, Suganya M, Suganthi Pandian, Sri Nithya Siva Sangara, Sharmilah M. Mogan, Suntharesan Siva Rajah, Maizun Binti Mohamad Ali Khan. Effects of Physical Activity on Body Mass Index among Medical Students from East Coast Peninsular Malaysian Public University. Research Journal of Pharmacy and Technology. 2023; 16(1): 200-4. doi: 10.52711/0974-360X.2023.00037

8. Koatz JG and Souza MDCB. Obese women and in vitro fertilization. JBRA Assisted Reproduction, 08 Aug 2022, 17(6): 353-356.

9. Baheerati M M, Gayatri Devi R. Obesity in relation to Infertility. Research J. Pharm. and Tech. 2018; 11(7): 3183-3185. doi: 10.5958/0974-360X.2018.00585.1

10. Revathi. R, Julius. A. A Biological Effect of Sex Hormone Binding Globulin and Testosterone in Polycystic Ovary Syndrome (PCOS) Obese Women. Research J. Pharm. and Tech. 2017; 10(7): 2143-2145

11. Mathyk BA1 and Quaas AM. Obesity and IVF: weighing in on the evidence. J Assist Reprod Genet. 2021 Feb; 38(2): 343–345.

12. Loveland JB, McClamrock HD, Malinow AM, Sharara FI. Increased body mass index has a deleterious effect on in vitro fertilization outcome. J Assist Reprod Genet. 2001; 18(7): 382–386.

13. Kumar R. Gautam GK. Pundir S. Zaidi S. Gupta C. Treatment of human infertility. Asian Journal of Research in Pharmaceutical Sciences. 2021; 11(2): 160-164. doi.org/10.52711/2231-5659.2021-11-2-12.

14. Bellver J , Ayllón Y, Ferrando M, Melo M, Goyri E, Pellicer A, Remohí J, Meseguer M. Female obesity impairs in vitro fertilization outcome without affecting embryo quality. Fertil Steril. 2010; 93(2): 447-454.

15. Koatz JG and Souza MDCB. Obese women and in vitro fertilization: results. JBRA Assisted Reproduction, 08 Aug 2022; 17(6): 353-356.

16. Banker M, Sorathia D, and Shah S. Effect of Body Mass Index on the Outcome of In-vitro Fertilization/Intracytoplasmic Sperm Injection in Women. J Hum Reprod Sci. 2017; 10(1): 37–43.

17. Shetty Sudeep Dinesh, Karunakar Hegde. Antiobesity activity of ethanolic extract of Citrus maxima leaves on cafeteria diet induced and drug induced obese rats. Research J. Pharm. and Tech. 2016; 9(7): 907-912.

18. Chaitali Bose, Alak Kumar Syamal, Koushik Bhattacharya. Pattern of Dietary Intake and Physical activity among Obese adults in Rural vs Urban areas in West Bengal: A Cross - Sectional Study. Research Journal of Pharmacy and Technology. 2022; 15(9): 3924-0. doi: 10.52711/0974-360X.2022.00657

19. Brewer CJ, Balen AH. The adverse effects of obesity on conception and implantation. Reproduction. 2010; 140:347–364.

20. Thomadakis C, Kramer B. . Effect of exogenous gonadotropins on gonadotrophs of the rat pituitary gland. The Anatomical Record. 1999; 254: 367–374

21. Edwards LJ, Kind KL, Amstrong DT and Thompsom JG..Effects of recombinant human follicle-stimulating hormone on embryo development in rat. American Journal of Physiology-Endocrinology and Metabolism.2005; 288(5): E845-E851.

22. Birowo P, Pujianto DA, Sahar N, Kusmardi K, Rusdi NK, Muharam R, Tjempakasari CR, Prasasty VD. Analysis of FSH-Receptor Expression in Testis of Infertile Men with Non-Obstructive Azoospermia (NOA). Research Journal of Pharmacy and Technology. 2024; 17(9): 4318-4. doi: 10.52711/0974-360X.2024.00667

23. Varghese, F., Bukhari, A. B., Malhotra, R., and De, A. IHC Profiler: An Open-Source Plugin for the Quantitative Evaluation and Automated Scoring of Immunohistochemistry Images of Human Tissue Samples. Plos One. 2014; 9(5): 1-11.

24. Rusdi NK, Purwaningsih EH, Hestiantoro A, Elya B, Kusmardi K. In Vivo Antimammary Tumor Effects of Soybean Extract with Targeted Lunasin (ET-Lun). Pharmacognosy Journal. 2021; 13(5): 1269-1276.

25. Valdez-Morales F. J., Gamboa-Domínguez A., Vital-Reyes V. S., et al. Changes in receptivity epithelial cell markers of endometrium after ovarian stimulation treatments: its role during implantation window. Reproductive health. 2015;12(1):1–11.

26. Liu G, Zang X, Lin H, Wang H, Li Q, Ni J, Zhu C. Effects of E-cadherin on mouse embryo implantation and expression of matrix metalloproteinase-2 and -9. Biochem Biophys Res Commun. 2006; May 12; 343(3): 832-838.

27. Angela Espir, Mohammad Y. Abajy, Ream Nayal. Investigating the effect of Pinus brutia bark on Pancreatic lipase and adiposity index in high-fat diet induced obese rats. Research Journal of Pharmacy and Technology. 2023; 16(4):1644-50.

28. Matsuzaki S, Darcha C, Maleysson E, Canis M, Mage G. Impaired Down-Regulation of E-Cadherin and β-Catenin Protein Expression in Endometrial Epithelial Cells in the Mid-Secretory Endometrium of Infertile Patients with Endometriosis. The Journal of Clinical Endocrinology and Metabolism. 2010; 95(7): 3437–3445

29. Jha RK, Titus S, Saxena D, Kumar PG, Laloraya M. Profiling of E-cadherin, β-catenin and Ca2+ in embryo-uterine interactions at implantation. 2006. FEBS;580: 5653-5660.

30. Best D, Avenell A, Bhattacharya. How effective are weight-loss interventions for improving fertility in women and men who are overweight or obese? A systematic review and meta-analysis of the evidence. Human Reprod Update, 2017; 1; 23 (6): 681-705.

31. Dakshita Snud Sharma, Sandip J Sutariya, Harmanpreet Kaur, Hitendra A Somani, Amit Gupta. Technological advancement: In vitro fertilization (IVF). Research Journal of Pharmacy and Technology. 2021; 14(12): 6721-4. doi: 10.52711/0974-360X.2021.01161

32. Hiba H. Kadhim, Salman A. Ahmed. Anti-mullerian Hormone and Vitamin D as a predictor of ovarian reserve and ovarian response in infertile women undergoing IVF. Research J Pharm and Tech 2019; 12(7): 3527-3530

33. Slater M, Murphy C, Barden J, Tenascin. E-cadherin and P2X calcium channel receptor expression is increased during rat blastocyst implantation. Histochem. J., 34, (2002), 13– 19.

34. Racca, A.; Bernabeu, A.; Bernabeu, R.; Ferrero, S. Endometrial receptivity in women with endometriosis. Best. Pract. Res. Clin. Obstet. Gynaecol. 2024, 92, 102438.

35. Li J, Zhang JV, Zhpu JK, Liu WM, Fan XJ, Duan EK. Inhibition of the β-catenin signaling pathway in blastocyst and uterus during the window of implantation in rat. Biol Reprod. 2005; 72: 700-706

36. Satterfield MC, Dunlap KA, HayashiK, BUrghard FW. Tight and adherens junctions in the ovine uterus: differential regulation by pregnancy and progesterone. Endocrinology. 2006; 148: 3911-3931.

37. Matsuzaki S, Darcha C, Maleysson E, Canis M, Mage G. Impaired down-regulation of E-Cadherin and beta-Catenin protein expression in endometrial epithelial cells in the mid-secretory endometrium of infertile patients with endometriosis.J. Clin.Endocrinol. Metab. 2010; 95: 3437-3445. doi:10.1210/jc.2009-2713.

38. Rahnama F, Shafiei F, Gluckman PD, Michel MD, Lobie PE. Epigenetic regulation of human trophoblastic cell migration and invasion. Endocrinol. 2006; 147: 5275-5283.

39. Alikani M. Epithelial cadherin distribution in abnormal human pre-implantation embryos. Human Reproduction, 20(12), 2005, 3369–3375.

Received on 11.11.2024 Revised on 19.05.2025

Accepted on 30.08.2025 Published on 01.12.2025

Available online from December 06, 2025

Research J. Pharmacy and Technology. 2025;18(12):6053-6058.

DOI: 10.52711/0974-360X.2025.00875

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Groups	Subgroups	Bodyweight	(gram)	E-Cadherin (H-score)	Expression	P value
Groups	Subgroups	Means	Ranges	Means	Ranges
Normal weight	Normal weight unstimulated (n=8)	179	165-181	179	143-217	-
	Normal weight stimulated (n=6)	142	169-180	142	114-173	0.07
	Overweight unstimulated (n=9)	264	250-283	226	202-244	0,02
	Overweight Stimulated group (n=7)	271	250-290	233	219-250	0.01