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PANCREATIC MORPHOMETRY UNDER STRESS
A.A.Umerov
Master's student, 1st year, “Morphology”
N.Q.Niyozov
Department of Human Anatomy and OSTA, PhD,
Tashkent Medical Academy, Tashkent, Uzbekistan
Abstract:
This study investigated the impact of maternal stress on pancreatic development
in rat offspring. Morphological analyses of the pancreas at various postnatal stages revealed
a developmental delay and altered acinar cell development in the experimental group
compared to controls. Notably, changes in both exocrine and endocrine cells were observed
from the third day of the experiment in all stressed animals, including destructive changes in
the mitochondria of acinocytes. Specifically, morphometric analysis showed a decrease of
up to 4% in pancreatic acini diameter and an 18% reduction in acini height in the
experimental group relative to controls. These hemomicrocirculatory changes, characterized
by interstitial stroma swelling and cellular dystrophic changes, were most pronounced on
day 14 and correlated with destabilization of acinocyte membrane structures. These findings
suggest that maternal stress induces significant morphological changes in offspring
pancreatic cells, potentially leading to chronic pancreatitis. The experimental design
involved subjecting pregnant rats to a stress-inducing maze environment, with offspring
pancreases examined histologically at 3, 7, 14, 21, and 30 days postnatal.
Keywords:
stress, pancreas, pancreatic lobes, acinus, morphological changes, experimental
stress, postnatal ontogenesis, exocrine cells, endocrine cells, rat model, maternal health,
chronic pancreatitis, histological preparation, morphometry, laboratory rats, saline solution,
experimental conditions, abdominal cavity.
Introduction.
Stress is a universal nonspecific neurohormonal response of the div in the
form of an increase in the div's resistance to damage or as a signal that threatens the life or
well-being of the organism. Today, stress disorders are one of the main problems of public
health concern. In addition, the COVID-19 pandemic and the consequences of social
isolation have had a profound impact on the mental health of society. These factors
constitute a heterogeneous cluster of mental disorders that can manifest as general physical
symptoms (for example, tachycardia, shortness of breath, or gastrointestinal disorders)
accompanied by anxiety. Other clinical symptoms associated with stress include excessive
or disproportionate anxiety and fear, persistent issues associated with impairment in social,
occupational, or other important areas of functioning.
Relevance of the work.
Gastrointestinal problems are commonly reported in studies of
occupational stress. Unfortunately, it is difficult to assess the physiological systems
underlying gastrointestinal symptoms in the workplace. Acute psychological stress has a
variable effect on pancreatic secretion, stimulating large increases in some individuals and
reducing production in others. Gastrointestinal problems are particularly high in many
workers, manifested by disruptions in the circadian rhythms of the central nervous system in
controlling pancreatic secretion. A growing div of research suggests that prenatal stress
can have significant effects on pregnancy, maternal health, and human development
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throughout an individual's life. These effects may occur directly through the impact of
prenatal stress-related physiological changes on the developing fetus or indirectly through
the effects of prenatal stress on maternal health and pregnancy outcomes, which in turn
affect infant health and development (The University of Colorado, Colorado Springs, and
The Colorado School of Public Health, 1420 Austin Bluffs Parkway, Colorado Springs, CO
80918, USA). The prevalence of stress in the United States has been found to have increased
by approximately 6.7% among individuals aged 18 to 25 from 2008 to 2018. The increasing
prevalence of anxiety and stress in society is considered to be the beginning of several
mental health problems, cardiovascular diseases, digestive diseases, and metabolic syndrome.
With changes in the environment and lifestyle, the incidence of pancreatic diseases has been
increasing year by year in recent years, and pancreatic diseases have become a common
ailment of the digestive system. The etiology of pancreatic diseases is diverse; the condition
is complex, treatment is relatively difficult, and the mortality rate is high. The harm of
pancreatic diseases to human health has attracted great attention from researchers. Reducing
the mortality rate and improving the prognosis of patients with pancreatic diseases is
becoming a major problem in medicine today.
Purpose:
To determine the nature of morphological and morphometric changes in the
pancreas under experimental stress.
Materials and methods of research.
To achieve the goal, 60 white laboratory rats were
subjected to experimental stress. The rats were divided into 2 groups. Group 1 is a control
group of healthy rats. Mother rats in the control group were given 1.0 ml of saline solution
every morning in the stomach. A subclavian catheter was used as a probe. Rat pups were
anesthetized under ether anesthesia on days 3, 7, 14, 21, and 30 after birth. Group 2 is an
experimental group, in which 50 female white laboratory rats were kept in specially
prepared maze cages to induce experimental stress. This stress model continued after the rats
became pregnant and gave birth. After opening the abdominal cavity of the animals, the
pancreas was removed, fixed in a 12% formalin solution, paraffin blocks were prepared, and
histological preparations were made from them. Experiments and slaughter of animals were
carried out in accordance with the "European Convention for the Protection of Vertebrate
Animals used for Experimental and other Scientific Purposes" (Strasbourg, 1985). At a
thickness of 8–10 μm, histological sections prepared on a rotary microtome were stained
with hematoxylin-eosin according to the standard method. Micropreparations were
photographed using an MBI-6 microscope. The obtained material was processed using the
methods of variation statistics (Boyarsky A.Ya. 1965; Kaminsky L.S. 1953; Sepetliev.D.
1968; Venchikov A.I.; Venchikov V.A.1974, G.G. Avtandilov, 1990) using the tables of
R.B. Strelkova (1986).
Results and Discussion.
The pancreas accounts for 2.45% of the total div weight of rats.
According to our studies, the pancreas of rats can be divided into three parts: 1. Duodenum -
located in the mesentery of the duodenum. 2. Biliary - located along the common bile duct. 3.
Stomach - located to the right of the spleen. The right lobe of the pancreas of rats is caudally
wrapped and is bordered by the duodenum, while the middle lobe surrounds the abdominal
wall and the cranial part of the mesenteric artery, and is limited by the transverse colon. The
left lobe, which has the shape of an elongated triangle, communicates with the spleen.
Externally, the pancreas is covered by a serous membrane and a connective tissue capsule of
the small intestine. In rats, the pancreas is located intraperitoneally. The exocrine (exocrine)
and intrasecretory (endocrine) parts are distinguished in the pancreas of rats. The delicate
stroma of the organ is represented by connective tissue, which divides the gland into lobes.
When viewed macroscopically, a connective tissue capsule surrounding it from all sides is
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revealed. From the capsule, a membrane penetrates deep into the organ, resulting in lobes
appearing in the parenchyma of the organ, giving the gland a lobed structure. The exocrine
part of the pancreas of rats forms a structural and functional unit - the acinus, the cross-
sectional area of which is (943.22±47.1) µm². The acinus consists of 8-12 exocrine
pancreatocytes or acinocytes. The average area of an exocrine pancreatocyte is (133.2 ± 2.5)
µm².
Morphological examination of the pancreas by the 14th day of the experiment showed that
changes in the vessels developed even more. An increase in edema was noted in the stroma,
mainly in the perivenular and pericapillary spaces (Fig. 2).
Collagen fibers are swollen and loose; the capsule is unevenly located, and swelling of
connective tissue along with thickening of trabeculae was clearly observed. The diameter of
the capillaries was 2.15±0.2 μm (1.88±0.056 μm in the control group), the walls were dilated,
and the stroma was filled with erythrocyte accumulation and tissue edema. It was found that
the diameter of the capillary blood vessels increased 1.2 times compared to the previous
group (Fig. 3).
Some of the acinocytes around the areas of hemorrhage were observed to be distant and
compressed, and the architecture of the islets was disturbed. In the peripheral parts of the
pancreas, inflammatory infiltration of the interacinar spaces between the acini and
hyperplasia of ductal epithelial cells were observed (Fig. 4).
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Significant structural changes occurred in endocrine cells. The average area of the nuclei
decreased, and karyopyknosis occurred. The area of α-cells decreased by 8.5% compared to
the control group. The α-cell nucleus showed enlarged pores and reduced electron density.
The area of secretory granules increased in α-cells, but the diameter of granules decreased
from 22.7±0.9 nm to 18.4±0.5 nm.
As a result of morphometric studies, the diameter of pancreatic acini is 11.4±0.3 μm, and
their height reaches 2.15±0.18 μm. Compared to the control group, acinar cell diameter and
height were delayed by 3% and 12% in the experimental group. These changes were evident
at 14 days.
On day 21, the segmental structure of the gland was preserved, but the volume of the
segments was slightly reduced. An increase in the intensity of swelling in the stroma of the
pancreas was observed, which spread throughout the gland and became diffuse. Swelling
and fragmentation of collagen fibers were observed in some areas. Reparative processes
were enhanced in the preserved parts of the stroma, with intensive proliferation of
fibroblasts and the formation of fibrils detected. The capillary diameter of the exocrine
parenchyma reached 2.24±0.26 μm (in the control group, it was 1.91±0.07 μm). The
capillary diameter increased by 1.3 times. In the exocrine cell acini, the location and
structure of the nucleus and nucleoli changed; cytoplasmic vacuolization, decreased
zymogen granules, and disturbed structure of the endoplasmic reticulum were noted. Loss of
degranulation was observed in the cytoplasm of endocrine cells. Interlobular venous vessels
were enlarged, leukocyte accumulation was detected, along with edema and myxomatous
changes on the vessel wall.
Plasmorrhagia was observed in the fibrous structures of the walls of small lobular vessels.
An increase in the number of degranulated eosinophils was found in the organ stroma.
As a result of morphometric studies, we found that the diameter of pancreatic acini is
11.7±0.2 μm, which is up to 4% compared to the control group, and their height is 2.2±0.15
μm, which is up to 18%.
The results of morphometric studies confirm that in hypothyroidism, the height of the
pancreatic gland acinar cells and their diameter gradually decrease compared to the control
group, indicating the development of an atrophic process in the gland parenchyma (See
Table 1).
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Table 1.
Age-related changes in pancreatic acinar cell morphometric parameters.
Rat children age (in
days) of
Atsinus cell diameter, µm Acinus cell height,
µm
3
10,7±0,2
1,9±0,1
7
11±0,25
2,05±0,15
14
11,4±0,3
2,15±0,18
21
11,7±0,2
2,2±0,15
30
11,9±0,2
*
2,3±0,2
*
Note:* - p<0.05 is reliable compared to the control group.
Conclusion.
Thus, changes are observed in the pancreatic acinocytes of offspring born to
mother rats under stress. These changes are hemomicrocirculatory in nature, manifested as
swelling of the interstitial stroma and destructive and dystrophic changes in the cells. These
changes are more pronounced on the 14th day of the experiment and are characterized by
destabilization of membrane structures in acinocytes. As a result, destructive changes are
detected in the mitochondria of acinocytes. Disorganization of acinocytes and their
structures indicates a violation of the contractile function of these cells. The data obtained
prove that changes in the pancreas at the cellular and subcellular levels lead to chronic
pancreatitis. As a result of morphometric studies, we observe that the diameter of the
pancreatic acini shows a decrease of up to 4%, and their height lags behind growth by up to
18% compared to the control group. Disruption of the architecture of the acinus was seen in
the sections located on the periphery of the pancreas. It can be observed that some
acinocytes around the blood-stained branches are separated and compressed.
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