دراســة نسجية كيموحيوية مناعية للدور الوقائي للمستخلص المائي والنانوي لجذور نبات الاشواغندا في أناث الجرذان المعاملة بمادة propyl4-hydroxybenzoate

A histological, immunohistochemical, and biochemical study on the protective role of the aqueous and nano extracts of Withania somnifera roots in female rats treated with propyl4-hydroxybenzoate

The present study aimed to evaluate the efficacy of aqueous extract and green nanocomposite of Ashwagandha root (Withania somnifera) against paraben-induced histopathological and histopathological changes of liver, kidney and ovarian tissues in female albino rats induced by 4-proplyhydroxybenzoate. The present study was conducted in the animal house of Faculty of Pharmacy / Karbala University from November 2023 to February 2024, 30 female albino rats were used and randomly divided into six groups with five animals per group, the first group (G1) was dosed with 1 ml/kg physiological solution and considered as control group, while the second group (G2) was dosed with 500 mg/kg bw aqueous extract of Ashwagandha root at a dose of 500 mg/kg bw. The third group (G3) was given the paraben 4-proplyhydroxybenzoate at 4.6 mg/kg. The fourth group was treated with a green nanocomposite of ashwagandha root at a dose of 150 mg/kg bw, the fifth group was treated with an aqueous extract of ashwagandha root and parabens at a dose of 500-4.6 mg/kg bw, and the sixth group was treated with a green nanocomposite of ashwagandha root and parabens at a dose of 150-4.6 mg/kg.
After 30 days, the experiment was terminated and 5 ml of blood was collected to measure the following parameters Hemoglobin (Hb), red blood cells (RBC), white blood cells (WBC), lymphocytes (LYM), white blood cells (GRA), oxidative stress index (MDA) malondialdehyde and some antioxidants such as glutathione (GSH) and superoxide dismutase (SOD) and levels of some female hormones E2 (estrogen) and progesterone (progesterone) and some immunological interleukins (IL-10 and IL-17).
The results of the present study showed that daily oral administration of parabens to female rats at a dose of 4.6 mg/kg resulted in a significant increase (P<0.05) in the levels of GRA, RBC, LYM, MDA, IL-10 and IL-17, and a significant decrease (P<0.05) in the levels of E2, estrogen and IL-17 compared to the control group.
The group that received the green nanocomposite at a dose of 150 mg/kg showed a significant increase (P<0.05) in the levels of GRA, RBC, WBC, LYM, SOD, and Progesterone compared to the control group. There was a significant decrease (P<0.05) in the level of IL-10 compared to the control group. There were no significant differences (P>0.05) in the levels of Hb MDA, GSH, E2, and IL-17 compared to the control group. The group dosed with aqueous extract of Ashwagandha root and parabens 4.6-500 mg/kg body weight showed a significant increase (P<0.05) in the levels of GRA, RBC, Hb, Progesterone, E2, IL-10 and IL-17 compared to the control group. There was a significant decrease (P<0.05) in the level of LYM compared to the control group. There were no significant differences (P>0.05) in the levels of MDA, GSH, E2, IL-17, WBC and SOD compared to the control group. The group that was dosed with green nanocomposite and parabens 4.6-150 mg/kg body weight showed a significant increase (P<0.05) in the levels of GRA, RBC, SOD, and Progesterone compared to the control group. There was a significant decrease (P<0.05) in the levels of MDA, GSH, and E2 compared to the control group. There were no significant differences (P<0.05) in the levels of Hb LYM, IL-10, IL-17, and WBC compared to the control group.
Histological examination of the kidney in the preservative-treated group at 4.6 mg/kg showed severe glomerul atrophy with damage to the walls of the urinary tubules compared with the control group. 6 mg/kg showed severe glomerul atrophy with damage to the walls of the urinary tubules compared with the control group, there was enlargement of Bowman’s space with severe vascul congestion, while the group treated with aqueous extract of ashwagandha root at 500 mg/kg and the group treated with nanocomposite at 150 mg/kg showed slight reduction in glomerul size, increase in Bowman’s space, normal Bowman’s capsule structure and regularity of proximal and distal convoluted tubules with no histopathological changes. The group treated with the extract and two hours later with the preservative showed a slight effect on the structure of the glomerulus and Bowman’s capsule, irregularity of the proximal convoluted tubule and distal convoluted tubule and an increase in Bowman’s space with severe congestion in the tubule area compared to the control group, while the group treated with the nanocomposite 150 mg/kg and two hours later with the preservative 4. 6 mg/kg, we observe normal shape of glomerulus G size, Bowman’s space, normal structure of Bowman’s capsule and regularity of proximal convoluted tubule and distal convoluted tubule compared to the control group and the group treated with aqueous extract of Ashwagandha root at a concentration of 500 mg/kg two hours later.
The preservative group and the group treated with the nanocomposite at a concentration of 150 mg/kg and two hours later also showed the normal shape of both proximal and distal urinary tubules and the normal shape of Bowman’s capsule, Bowman’s space and glomerulus compared to the preservative group. Histological changes in the liver tissue of rats at 4.6 mg/kg preservative showed congestion of the central vein, nucleoli, sinusoidal dilatation, presence of chronic inflammatory cells around the vein, presence of nucleoli and irregularity of hepatic cords compared with the control group. As for the group treated with aqueous extract of Ashwagandha root at a concentration of 500 mg/kg and the group treated with nanocomposite at a concentration of 150 mg/kg, the histological structure of the liver appeared close to normal tissues, consisting of the central vein and the regularity of hepatic cords consisting of ribbed hepatocytes and spherical nuclei with the presence of hepatic sinusoids as compared to the control group. The results of liver histology of rats treated with Ashwagandha extract 500 mg/kg and two hours later administered with preservative 4. The histological structure of the liver appeared close to normal tissues as it consists of the central vein with the presence of hepatic sinusoids and the hepatic cords represented by hepatocytes are polygonal and the nuclei are spherical We note the presence of slight congestion in the cells when compared to the control group. The histological results of the liver of the group of rats treated with the nanocomposite at 150 mg/kg and two hours later with a preservative at 4.6 mg/kg showed the presence of chronic inflammatory cells around the vein, showing the normal shape of liver cells with their regul hexagonal shape and regularity in the sinusoids. These changes, which were almost normal in the liver tissue in both groups and compared with the control group, while the group dosed with ashwagandha root extract 500 mg/kg and two hours later preservative 4.6 mg/kg and the group dosed with nanocomposite 150 mg/kg and two hours later preservative 4.6, we observed the normal shape of liver cells with regul hexagonal shape and regularity in the sinusoids compared with the preservative group. As for the changes in the ovarian tissue of rats treated with preservative 4.6 mg/kg, we observed that the ovarian tissue was affected by the appearance of areas of inflammation in addition to the appearance of ovarian follicles at different stages, but fewer in number and less cystic compared to the control, while the group treated with aqueous extract of Ashwagandha root 500 mg/kg and two hours later preservative 4.6 mg/kg and the nanocomposite group were affected by the appearance of areas of inflammation. 6 mg/kg and the nanocomposite group 150 mg/kg and two hours later preservative 4.6 mg/kg, we observed the appearance of ovarian tissue appearing normal and the appearance of ovarian follicles well, as well as in large number and in their various forms as well as the appearance of corpus luteum compared to the control group.