The ovary is not only a reproductive organ but also a dynamic endocrine tissue whose function is closely coordinated with the hypothalamus and pituitary gland. Through the hypothalamic–pituitary–ovarian (HPO) axis, pulsatile gonadotropin-releasing hormone regulates pituitary secretion of follicle-stimulating hormone (FSH) and luteinizing hormone (LH), which in turn govern follicular development, ovulation, luteal function, and ovarian steroidogenesis [1]. Ovarian hormones subsequently provide feedback to the hypothalamic–pituitary system, creating a finely balanced endocrine network that changes throughout the reproductive lifespan [2]. Within this axis, FSH primarily supports follicular recruitment and maturation, whereas LH contributes to follicular steroidogenesis, ovulation, and maintenance of luteal function [1]. Progesterone (P4) is a major ovarian steroid, produced predominantly by the corpus luteum following ovulation, and its circulating concentration varies considerably with menstrual-cycle phase and reproductive age [2]. Prolactin (PRL), although best known for its role in lactation, also interacts with reproductive physiology and has been implicated in follicular function, corpus luteum activity, and regulation of fertility [3]. Consequently, interpretation of these hormones requires consideration of age, ovarian function, and reproductive status. Benign ovarian tumors comprise histologically diverse lesions arising from epithelial, germ-cell, or sex cord–stromal components. While many benign ovarian tumors are not regarded as classical hormone-producing lesions, certain tumors particularly those arising from sex cord and stromal tissues may retain steroidogenic activity and present with hormone-related manifestations [4]. Even in the absence of overt endocrine symptoms, the presence of ovarian pathology may coexist with variations in the hormonal environment. At the same time, reproductive aging itself has an important influence: progressive follicular depletion is accompanied by altered ovarian feedback, increasing gonadotropin concentrations and declining ovarian steroid production, particularly across the menopausal transition [5]. These age-dependent changes make an age-comparable control population particularly important when endocrine profiles are examined. The endocrine activity of specific hormone-secreting ovarian tumors is well recognized; however, the broader pattern of circulating reproductive hormones across a heterogeneous population of benign ovarian tumors remains less clearly characterized. Most available literature has focused on individual functional ovarian tumors, fertility-related endocrine disorders, or malignant ovarian disease rather than simultaneously examining conventional reproductive hormones in benign ovarian pathology [4,5]. Therefore, the present study aimed to compare serum LH, FSH, P4, and PRL concentrations between women with benign ovarian tumors and age-matched healthy controls. It further evaluated the relationships of these hormones with age and with one another and assessed their ability to discriminate women with benign ovarian tumors from healthy controls.

Study Design and Participants This case-control study was conducted at a tertiary care center in Miraj, Maharashtra and included 100 women. Of these, 50 had benign ovarian tumors and 50 were age-matched healthy controls. The study was structured and reported in accordance with recommendations for observational case-control studies [6]. The two groups and their sample sizes correspond to the final study dataset. Inclusion Criteria Women with documented benign ovarian pathology, availability of relevant clinical information, complete serum measurements of luteinizing hormone (LH), follicle-stimulating hormone (FSH), progesterone (P4), and prolactin (PRL), and informed consent were included in the study group. The control group comprised age-matched healthy women without evidence of malignancy or significant systemic illness likely to influence the biochemical parameters under investigation and with adequate clinical and laboratory data. Exclusion Criteria Women with a history of smoking, evidence of malignancy, significant systemic illness likely to influence hormonal parameters, severe hepatic, cardiac, or renal disease, incomplete clinical or laboratory data, or unwillingness to provide informed consent were excluded. Clinical and Pathological Assessment Relevant demographic and clinical information was recorded for all participants. Age was documented in both groups, while the site of ovarian involvement was recorded in women with benign ovarian tumors. The benign group included a spectrum of ovarian lesions, and classification as benign was based on the pathological diagnosis documented in the study records. Hormonal Assessment Serum concentrations of LH, FSH, P4, and PRL were measured using standard immunoassay methods with commercially available reagents and laboratory platforms. All hormonal measurements were performed according to the manufacturers recommended analytical procedures and routine laboratory quality-control practices. Statistical Analysis Continuous variables were summarized as mean ± standard deviation or median with interquartile range, as appropriate, while categorical variables were expressed as frequencies and percentages. Because the serum hormone concentrations were analyzed using non-parametric methods, differences between women with benign ovarian tumors and age-matched healthy controls were assessed using the Mann–Whitney U test. Relationships between age and serum hormone concentrations, as well as inter-hormonal associations, were evaluated using Spearman’s rank correlation coefficient (ρ). Receiver operating characteristic (ROC) curve analysis was performed to examine the ability of individual hormones to discriminate women with benign ovarian tumors from healthy controls. The area under the ROC curve (AUC), optimal cutoff value, sensitivity, and specificity were determined [7]. A two-sided p-value <0.05 was considered statistically significant. Ethical Considerations The study was conducted after approval from the relevant Institutional Ethics Committee. Written informed consent was obtained from all participants before enrolment. Participant identity and clinical and laboratory information were kept confidential throughout data collection, analysis, and reporting.

Table 1: Demographic and clinical characteristics of the study participants Variable Age-Matched Healthy Controls (n=50) Benign (n=50) Age (years) Mean ± SD 36.90 ± 17.89 38.74 ± 14.63 Median (IQR) 30.50 (23.75–50.25) 37.00 (25.00–47.75) Min–Max 12–80 17–80 Age group n (%) ≤20 4 (8.0) 4 (8.0) 21–30 21 (42.0) 12 (24.0) 31–40 10 (20.0) 14 (28.0) 41–50 3 (6.0) 12 (24.0) 51–60 4 (8.0) 4 (8.0) >60 8 (16.0) 4 (8.0) Site of ovary n (%) Right - 29 (58.0) Left - 18 (36.0) Bilateral - 2 (4.0) Mass - 1 (2.0) Total 50 (100.0) 50 (100.0) Continuous variables are presented as mean ± SD, median (IQR), and minimum–maximum. Categorical variables are presented as n (%). A total of 100 participants were included in the study, comprising 50 age-matched healthy controls and 50 patients with benign ovarian tumors. The mean age of the age-matched healthy controls was 36.90 ± 17.89 years (table 1), with a median age of 30.50 years (IQR: 23.75–50.25) and an age range of 12–80 years. Among patients with benign ovarian tumors, the mean age was 38.74 ± 14.63 years, with a median age of 37.00 years (IQR: 25.00–47.75) and an age range of 17–80 years. Among the age-matched healthy controls, the majority of participants were in the 21–30-year age group (42.0%), followed by those aged 31–40 years (20.0%) and >60 years (16.0%). The ≤20-year and 51–60-year groups each accounted for 8.0%, while the 41–50-year group represented 6.0% of the controls. Among patients with benign ovarian tumors, the largest proportion belonged to the 31–40-year age group (28.0%), followed by the 21–30-year and 41–50-year age groups (24.0% each), while participants aged ≤20 and 51–60 years each represented 8.0%, and those aged >60 years represented 8.0%. Regarding the site of ovarian involvement among the 50 patients with benign ovarian tumors, the right ovary was the most frequently involved site (29, 58.0%), followed by the left ovary (18, 36.0%). Bilateral involvement was observed in 2 (4.0%) patients, while 1 (2.0%) patient was recorded as having a mass. Table 2: Comparison of Serum Hormone Levels Between Age-Matched Healthy Controls and Patients with Benign Ovarian Tumors Hormone Age-Matched Healthy Controls Benign p-value LH 20.75 (8.83–48.25) 15.30 (11.28–21.45) 0.741 FSH 13.30 (8.38–37.88) 9.15 (6.68–18.55) 0.103 P4 1.15 (0.68–7.35) 4.80 (0.88–15.10) 0.045* PRL 10.15 (6.18–17.45) 12.40 (6.68–17.45) 0.605 Values are presented as median (IQR). Mann–Whitney U test was used; p < 0.05 was considered statistically significant. The serum hormone levels were compared between age-matched healthy controls (n = 50) and patients with benign ovarian tumors (n = 50) using the Mann–Whitney U test. The median LH level was 20.75 mIU/mL (IQR: 8.83–48.25) in age-matched healthy controls and 15.30 mIU/mL (IQR: 11.28–21.45) in the benign group; however, the difference was not statistically significant (p = 0.741) (table 2). Similarly, the median FSH level was 13.30 mIU/mL (IQR: 8.38–37.88) among healthy controls and 9.15 mIU/mL (IQR: 6.68–18.55) among patients with benign ovarian tumors, with no statistically significant difference (p = 0.103). In contrast, progesterone levels differed significantly between the groups. The median progesterone level was 1.15 ng/mL (IQR: 0.68–7.35) in healthy controls compared with 4.80 ng/mL (IQR: 0.88–15.10) in the benign ovarian tumor group (p = 0.045), with higher levels observed in the benign group. The median prolactin level was 10.15 ng/mL (IQR: 6.18–17.45) in healthy controls and 12.40 ng/mL (IQR: 6.68–17.45) in the benign group. This difference was not statistically significant (p = 0.605). Overall, among the four hormones evaluated, only progesterone showed a statistically significant difference between age-matched healthy controls and patients with benign ovarian tumors. Table 3: Correlation Analysis of Age and Serum Hormone Variables in Benign Ovarian Tumors Variable 1 Variable 2 Spearman's ρ p-value Age LH 0.634 <0.001* Age FSH 0.713 <0.001* Age P4 −0.470 0.001* Age PRL −0.056 0.699 LH FSH 0.75 <0.001* LH P4 −0.379 0.007* LH PRL 0.255 0.073 FSH P4 −0.500 <0.001* FSH PRL 0.137 0.343 P4 PRL 0.121 0.401 Spearman's rank correlation was used. A p-value <0.05 was considered statistically significant. Spearman’s rank correlation analysis was performed to assess the relationships between age and serum hormone levels, as well as the associations among the individual hormones in patients with benign ovarian tumors. A strong positive correlation was observed between age and FSH (ρ = 0.713, p < 0.001), while age also showed a strong positive correlation with LH (ρ = 0.634, p < 0.001) (table 3). In contrast, age demonstrated a moderate negative correlation with progesterone (ρ = −0.470, p = 0.001). No significant correlation was observed between age and prolactin (ρ = −0.056, p = 0.699). Among the hormones, LH and FSH showed a strong positive correlation (ρ = 0.750, p < 0.001). LH showed a weak-to-moderate negative correlation with progesterone (ρ = −0.379, p = 0.007), while FSH demonstrated a moderate negative correlation with progesterone (ρ = −0.500, p < 0.001). No statistically significant correlations were observed between LH and prolactin (ρ = 0.255, p = 0.073), FSH and prolactin (ρ = 0.137, p = 0.343), or progesterone and prolactin (ρ = 0.121, p = 0.401). Overall, age was positively associated with LH and FSH and negatively associated with progesterone, while LH and FSH were strongly positively correlated with each other. Table 4: Diagnostic Performance of Serum Hormones for Discriminating Benign Ovarian Tumors from Age-Matched Healthy Controls Hormone AUC Cutoff Sensitivity (%) Specificity (%) LH 0.481 15.35 50 48 FSH 0.405 13.3 38 50 P4 0.616 1.15 74 50 PRL 0.53 10.05 62 50 ROC analysis demonstrated that progesterone showed the highest discriminatory ability among the four serum hormones, with an AUC of 0.616 (95% CI: 0.503–0.730; p = 0.045). At a cutoff of 1.15 ng/mL, progesterone showed 74.0% sensitivity and 50.0% specificity. LH (AUC = 0.481, p = 0.741), FSH (AUC = 0.405, p = 0.103), and PRL (AUC = 0.530, p = 0.605) (table 4) showed poor discriminatory ability and were not statistically significant. Overall, progesterone demonstrated only limited discriminatory performance, while the other hormones showed little ability to distinguish benign ovarian tumors from age-matched healthy controls. Figure 1: Comparison of Serum Hormone Levels Between Age-Matched Healthy Controls and Benign Ovarian Patients Figure 1: Comparison of Serum Hormone Levels Between Age-Matched Healthy Controls and Benign Ovarian Patients The box plots showed differences in serum hormone levels between age-matched healthy controls and benign ovarian patients. Median FSH and LH levels were higher in healthy controls (13.30 and 20.75, respectively) than in benign ovarian patients (9.15 and 15.30, respectively). In contrast, median P4 and PRL levels were higher in benign ovarian patients (4.80 and 12.40, respectively) than in healthy controls (1.15 and 10.15, respectively).

The present study evaluated serum LH, FSH, P4, and PRL in women with benign ovarian tumors compared with age-matched healthy controls. As shown in Table 1, the two groups had comparable overall age profiles, while most benign lesions were unilateral and right-sided. This pattern is consistent with the report of Kondi-Pafiti et al. (2023), who evaluated 1,355 women with histologically confirmed benign ovarian tumors and also observed a tendency toward right-sided predominance in some benign histological types [8]. The major endocrine finding was that P4 was the only hormone showing a significant between-group difference (Table 2). Median P4 was 4.80 ng/mL in women with benign ovarian tumors compared with 1.15 ng/mL in controls (p=0.045), whereas LH, FSH, and PRL did not differ significantly. Yang et al. (2023) studied 150 women with benign ovarian tumors and 104 healthy controls and similarly found no significant differences in LH or FSH between the groups [9]. This supports the present observation that benign ovarian pathology does not necessarily produce a generalized disturbance in circulating gonadotropins. The higher P4 concentration suggests that some benign ovarian lesions may be associated with altered ovarian steroidogenic activity. Hormonal behaviour, however, may vary considerably according to histological subtype. Matsuoka et al. (2022) demonstrated marked differences in LH, FSH, estradiol, and progesterone across ovarian tumor types, with progesterone being significantly higher in hormonally active granulosa-cell tumors than in other ovarian tumors [10]. Although their population differs from ours, the study illustrates that ovarian pathology can influence circulating steroid profiles. Therefore, the P4 elevation observed here should be interpreted cautiously, particularly because menstrual-cycle phase and menopausal status were not separately analysed. Age-related relationships were prominent (Table 3). Age correlated positively with LH (ρ=0.634) and FSH (ρ=0.713) and negatively with P4 (ρ=−0.470). Soares et al. (2020), in a longitudinal cohort of 1,608 women, similarly demonstrated increasing LH and FSH across the menopausal transition [11]. Santoro et al. (2020) further showed declining luteal progesterone as women approached the final menstrual period [12]. These observations support reproductive aging as an important contributor to the hormonal pattern seen in our study. The strong LH–FSH correlation also reflects their coordinated regulation within the hypothalamic–pituitary–ovarian axis. PRL showed neither a significant between-group difference nor meaningful correlations. This is relevant because Ganie et al. (2023) demonstrated substantial physiological variation in LH, FSH, and PRL among healthy Indian women and emphasized the influence of age and physiological state when interpreting reproductive hormones [13]. Finally, Table 4 and Figure 1 showed that P4 had the highest AUC (0.616), but its 74% sensitivity and 50% specificity indicate only limited discriminatory ability. LH, FSH, and PRL performed close to chance. Thus, these hormones are more informative for understanding the endocrine context of benign ovarian pathology than for independent diagnosis. The study is strengthened by its age-matched design and simultaneous hormonal assessment. Limitations include modest sample size, heterogeneous tumor types, wide age range, and lack of stratification by menstrual phase and menopausal status. A key strength of the present study was the simultaneous evaluation of LH, FSH, P4, and PRL in women with benign ovarian tumors using an age-matched healthy control group. In addition to between-group comparisons, correlation analysis and ROC assessment provided a broader understanding of both endocrine relationships and the limited discriminatory value of these hormones in benign ovarian pathology.

Benign ovarian tumors were associated with selective endocrine alterations rather than a generalized hormonal disturbance. Among the hormones studied, P4 was significantly higher in women with benign ovarian tumors, whereas LH, FSH, and PRL did not differ significantly from age-matched healthy controls. Age showed strong positive associations with LH and FSH and an inverse association with P4, highlighting the important influence of reproductive aging on endocrine interpretation. Although P4 showed the highest ROC performance, its discriminatory ability was limited and does not support its use as an independent diagnostic marker. Overall, these findings suggest that serum hormonal profiles may provide useful insight into the endocrine environment associated with benign ovarian pathology, but their interpretation should remain integrated with age, reproductive status, clinical findings, and pathological diagnosis.

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