Diabetes mellitus is a chronic disease, with a marked impact on quality of life, considered one of the most important and common diseases encountered in the medical clinic and which at the present time, due to the multiple complications and comorbidities involved in this pathology, represents a real risk at the global level [1]. Diabetes and prediabetic conditions are often presenting features of previously recognised endocrine disorders. Further, diabetes is another comorbidity that increases cardiovascular mortality but is also a modifiable and individualizing characteristic in a patient. More often than not diabetes secondary to endocrine diseases can be well controlled if the hormone imbalance is well treated [2]. Type II diabetes a chronic metabolic disorder that leads to hyperglycemia as a result of insulin resistance or inadequate insulin supply has become prominent prompting researcher to look into more details concerning the disease [3]. DA however affects metabolism and the glucose levels and as the disease progresses a series of complications occur affecting organs and tissues in the body. Such the relationship between histopathological features microscopic changes in tissues and hormonal profile in patients with Type II diabetes is the issue of considerable importance that provides insights into the development of the diabetes and its possible treatment [4].

Pathology analysis of tissues gives an insight into the effects of diabetes on the human organs in relation to cellular as well as tissue structures. Organizationally, special damage is incurred in the Type II diabetes where the high levels of glycemia remain elevated for longer periods of time and cause substantial changes in the tissues of the body especially in the pancreas, kidneys, liver and other components of the body associated with glycemic regulation [5]. Some of these alterations are such as the alteration in layer thickness where the bassement membrane may increase in thickness or become fibrotic or show the accumulation of advance glycation end products (AGEs), which are proteins or lipids that get glycated following exposure to sugars [6]. The following alterations help in the development of complications of diabetes such as nephropathy, retinopathy, and cardiovascular diseases. This factor if not diagnosed correctly can lead to Diabetes that results from an endocrine disorder being estimated as type 2 diabetes [7]. The ADA classification of diabetes includes four categories: First, there is type 1 diabetes, which is caused by the autoimmune destruction of pancreatic β cells, and absolute insulin deficiency; the second type is characterized by a progressive decline of endogenous insulin secretion on the background of insulin resistance; gestational diabetes and other specific types of diabetes are also distinguished based on the underlying causes: hereditary defects of pancreatic β cell activity and insulin sensitivity, diseases of the exocrine pancreas, endocrine disorders, drugs [8,9].

Objective

The main objective of the study is to find the correlation between histopathological features and hormonal profile in type II diabetes patients.

This correlational study was conducted at Services Institute of Medical Sciences, Lahore during June 2023 to March 2024. Data were collected from 185 diabetic patients.

Patient Selection

The inclusion criteria for the study were as follows:

• Patients aged between 40 and 70 years.
• Diagnosed with Type II diabetes for at least five years.
• Currently under medical treatment for diabetes, including oral hypoglycemic agents or insulin therapy.
• No history of other major chronic illnesses such as cancer or chronic kidney disease that could potentially affect the study results.

Patients were excluded if they had:

• A history of Type I diabetes.
• Severe diabetic complications such as advanced diabetic nephropathy or retinopathy.
• Any recent surgeries or significant medical interventions that could influence hormonal profiles or histopathological features.

Data Collection

The study involved the collection of both histopathological and hormonal data from the patients. The following procedures were undertaken:

Histopathological Analysis:

• Biopsies were taken from target organs, including the pancreas, liver, and kidneys, which are commonly affected in Type II diabetes.
• Tissue samples were fixed in formalin, embedded in paraffin, and sectioned for microscopic examination.
• The sections were stained using hematoxylin and eosin (H&E) as well as special stains such as Periodic acid-Schiff (PAS) and Masson's trichrome to highlight specific histopathological features.
• Histopathological features assessed included:
• Pancreatic islet cell morphology, including the degree of beta-cell depletion.
• Presence and extent of fibrosis in the liver and kidneys.
• Thickening of the basement membrane in renal tissues.
• Accumulation of advanced glycation end products (AGEs).
• All samples were analyzed by two independent pathologists to ensure consistency and reliability in the assessment.

Hormonal Profile Assessment:

• Blood samples were collected from patients after an overnight fast.
• Serum levels of key hormones were measured, including:
• Fasting insulin and C-peptide levels to assess insulin secretion and resistance.
• Glucagon levels to evaluate the counter-regulatory response to insulin.
• Adipokines such as leptin and adiponectin to explore the role of adipose tissue in metabolic regulation.
• Cortisol levels to assess the impact of stress hormones on glucose metabolism.
• Hormone levels were measured using enzyme-linked immunosorbent assay (ELISA) kits, following the manufacturer’s protocols.

Statistical Analysis:

The data collected were entered into a statistical software package v23 for analysis. Descriptive statistics were used to summarize the demographic and clinical characteristics of the patients.

The study included 185 patients with Type II diabetes, comprising 98 males (53%) and 87 females (47%). The average age of participants was 55.23 years, with a standard deviation of 3.56 years. On average, patients had been living with diabetes for 8.5 years. The mean BMI was 28.5 kg/m², with a range between 22 and 35 kg/m², indicating that most participants were overweight or borderline obese.

Table 1: Demographic and Clinical Characteristics of Patients

The analysis of tissue types revealed that 75% of patients experienced beta-cell depletion in the pancreas, with an average severity of 6.5 on a scale of 1-10. Abnormal islet cell morphology affected 68% of patients, with a mean severity of 5.0. Liver fibrosis and steatosis were observed in 58% and 64% of patients, with severity scores of 4.8 and 5.6, respectively. In the kidneys, 72% of patients showed basement membrane thickening, while 65% exhibited glomerular hypertrophy, with mean severities of 6.2 and 5.4. Additionally, 70% of patients had accumulation of advanced glycation end-products (AGEs) across multiple organs, with a mean severity of 6.7.

Table 2: Histopathological Findings

The hormone analysis showed that 60% of patients had abnormal fasting insulin levels, with a mean value of 20.5 µU/mL, exceeding the normal range of 2-20 µU/mL. C-Peptide levels were elevated in 52% of patients, with a mean of 3.2 ng/mL compared to the normal range of 0.5-2.0 ng/mL. Glucagon levels were abnormal in 55% of patients, averaging 200 pg/mL, the upper limit of the normal range (50-200 pg/mL). Leptin levels were elevated in 65% of patients, with a mean of 25 ng/mL, far above the normal range of 5-15 ng/mL. Additionally, 62% of patients had abnormal adiponectin levels, with a mean of 4.5 µg/mL, which is below the expected normal range of 5-30 µg/mL. Cortisol levels were abnormal in 56% of patients, with a mean value of 18 µg/dL within the normal range of 10-30 µg/dL.

Table 3: Hormonal Profile

The correlation analysis revealed a strong negative correlation between beta-cell depletion and fasting insulin (r = -0.68, p < 0.001), indicating that as beta-cell depletion increases, fasting insulin levels significantly decrease. There was a moderate negative correlation between liver fibrosis and adiponectin (r = -0.52, p < 0.01), suggesting that greater fibrosis is associated with lower adiponectin levels. A strong positive correlation was observed between kidney basement membrane thickening and glucagon (r = 0.63, p < 0.001), indicating that increased thickening corresponds with higher glucagon levels. The accumulation of AGEs and cortisol showed a moderate positive correlation (r = 0.58, p < 0.01), implying that higher AGE accumulation is associated with elevated cortisol levels.

Table 4: Correlation Analysis Between Histopathological Features and Hormonal Profile

The data indicates that as the duration of diabetes increases, the prevalence of complications also rises. For patients with diabetes lasting 5-7 years, 58% experienced beta-cell depletion, 43% had liver fibrosis, 55% showed kidney basement membrane thickening, and 60% had accumulation of AGEs. Among those with 8-10 years of diabetes, the prevalence of these conditions increased to 77%, 65%, 75%, and 72%, respectively. For patients with more than 10 years of diabetes, the percentages further escalated to 89% for beta-cell depletion, 82% for liver fibrosis, 87% for kidney basement membrane thickening, and 85% for AGE accumulation.

Table 5: Distribution of Histopathological Features by Duration of Diabetes

The study explored the correlation between histopathological features and hormonal profiles in patients with Type II diabetes, providing valuable insights into the complex interplay between cellular changes and hormonal imbalances in the progression of the disease. The findings of this study have several important implications for understanding the pathophysiology of Type II diabetes and guiding future research and clinical practice [11]. The results indicate that significant histopathological changes occur in key organs such as the pancreas, liver, and kidneys in patients with Type II diabetes. Notably, beta-cell depletion in the pancreas was observed in a large proportion of patients, particularly those with a longer duration of diabetes [12]. This finding aligns with the well-established understanding that beta-cell dysfunction and loss are central to the progression of Type II diabetes, contributing to impaired insulin secretion and the eventual need for insulin therapy. The observed correlation between beta-cell depletion and increased insulin resistance further underscores the role of beta-cell failure in the pathogenesis of the disease [13].

Liver fibrosis and steatosis were also prevalent among the study participants, highlighting the impact of Type II diabetes on liver health. The development of fibrosis in the liver can be attributed to chronic hyperglycemia and insulin resistance, which promote inflammation and the deposition of extracellular matrix proteins. The negative correlation between liver fibrosis and adiponectin levels suggests that reduced adiponectin, a hormone with anti-inflammatory and insulin-sensitizing properties, may contribute to the progression of hepatic fibrosis in diabetic patients [14]. This finding is particularly significant, as it points to adiponectin as a potential therapeutic target for preventing or mitigating liver damage in diabetes. In the kidneys, the thickening of the basement membrane and glomerular hypertrophy were common, indicating early signs of diabetic nephropathy [15]. The strong correlation between these renal histopathological changes and elevated glucagon levels suggests that dysregulation of glucagon, a hormone that raises blood glucose levels, may play a role in the development of kidney damage in diabetes. This finding is consistent with emerging evidence that hyperglucagonemia is a key factor in the metabolic disturbances seen in Type II diabetes, and it highlights the need for further research into glucagon as a therapeutic target [16].

The accumulation of advanced glycation end products (AGEs) in multiple tissues, particularly in the kidneys and liver, was another significant finding. AGEs are known to contribute to tissue damage through mechanisms such as oxidative stress and inflammation. The positive correlation between AGE accumulation and cortisol levels suggests that elevated stress hormones may exacerbate AGE formation and its deleterious effects on tissues. This relationship between cortisol and AGEs underscores the importance of managing stress and reducing oxidative stress in patients with Type II diabetes to prevent complications [17]. The study also found that hormonal imbalances, particularly hyperinsulinemia, elevated glucagon levels, low adiponectin levels, and high leptin levels, were prevalent among the patients, with a clear association between these abnormalities and higher BMI. These findings highlight the critical role of obesity in exacerbating hormonal dysregulation in Type II diabetes. Obesity, characterized by excess adipose tissue, is known to alter the secretion of adipokines, such as leptin and adiponectin, leading to insulin resistance and increased risk of metabolic complications [18].

The association between higher BMI and hyperinsulinemia indicates that insulin resistance is more pronounced in obese patients, likely due to the increased demand on pancreatic beta-cells to produce insulin. The observation that patients with higher BMI also had elevated glucagon levels suggests that obesity may exacerbate the dysregulation of glucagon secretion, further contributing to hyperglycemia. This finding supports the growing body of evidence that targeting both insulin and glucagon pathways may be necessary for effective diabetes management, particularly in obese individuals [19]. Low adiponectin levels in obese patients were associated with more severe liver fibrosis, indicating that adiponectin deficiency may play a key role in the development of non-alcoholic fatty liver disease (NAFLD) and its progression to non-alcoholic steatohepatitis (NASH) in Type II diabetes. Since adiponectin has protective effects on liver tissue, enhancing adiponectin levels through lifestyle interventions or pharmacological agents could be a promising strategy for preventing liver complications in obese diabetic patients [20]. The findings of this study have several clinical implications for the management of Type II diabetes. First, the identification of strong correlations between histopathological features and hormonal profiles suggests that regular monitoring of both tissue changes (e.g., through biopsy or imaging) and hormonal levels may provide a more comprehensive understanding of disease progression. This could lead to more personalized treatment approaches that address both the underlying cellular damage and the hormonal imbalances in patients with Type II diabetes.

It is concluded that significant correlations exist between histopathological features and hormonal profiles in Type II diabetes patients, with beta-cell depletion, liver fibrosis, and kidney damage closely linked to insulin resistance, glucagon dysregulation, and adiponectin deficiency.

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