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. 2023 Jul 19;11(7):625.
doi: 10.3390/toxics11070625.

Immunological and Histological Studies of Different Concentrations of Rosmarinus officinalis and Thymus vulgaris Extracts on Thymus Gland of Chick Embryos

Reem Yahya Alzahri  1 , Fawzyah Abdullah Al-Ghamdi  1 , Seetah Saleem Al-Harbi  1
Affiliations

Immunological and Histological Studies of Different Concentrations of Rosmarinus officinalis and Thymus vulgaris Extracts on Thymus Gland of Chick Embryos

Reem Yahya Alzahri et al. Toxics. .

Abstract

Humanity has an ancient history of consuming medicinal plants for prophylaxis. Within hours, and even months, embryonic cells undergo several processes to form an organism. This study aimed to prove the positive or negative effects of using rosemary and thyme extract on the thymus gland and level of IL-10, IgM, and IgG in serum of chick embryos. The immunological effect was measured by histological and immunological studies. A total of 160 fertilized eggs were randomly distributed into 8 groups; on the 0 and 8th day of incubation, all treated groups received a dose of 0.1 mL/egg. On the 14th and 20th days of incubation, the embryos were sacrificed and the samples were collected (serum and thymus gland). The data were analyzed using ANOVA. Simple damage in thymic tissue with a low cell density in the embryos was treated with high concentrations of rosemary and thyme extracts, as well as in the mixed group. A significant decrease in IgM levels in the group treated by a high concentration of thyme. A decrease in IgG levels was found in the group treated with a high concentration of rosemary and in the mixed group, while the group treated with a high concentration of thyme and the mixed group showed decreases on the 14th day. A significant decrease in IL-10 levels was found on the 14th day, followed by an increase on the 20th day. Despite the benefits of rosemary and thyme, inflammation signs appeared on embryos treated with these herbs.

Keywords: IL-10; IgG; IgM; embryo developments; rosemary; thyme; thymus gland.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Showing the growth in the thymus gland over time. In Figure (A,B) the thymus appears during the second week of incubation (14th day), and in Figure (C), in the last week of incubation (20th day), an accurate structure of the thymus gland appears; which was encapsulated by a capsule (CP) and the lobules separated by septa (S); inside the gland, the cells organized and distributed in the cortex © and medulla (M). H&E stain. Scale bar was 1 mm, 50 µm, and 200 µm, respectively.
Figure 2
Figure 2
Shows the cortex of the thymus gland. It is darkly stained and abundant numbers of lymphocytes appear (yellow arrow). H&E stain. Scale bar 20 µm.
Figure 3
Figure 3
Shows the medulla of the thymus gland. There are several mature lymphocytes (yellow arrow) and a high density of epithelial cells with pale stained nuclei (red circle). H&E stain. Scale bar 20 µm.
Figure 4
Figure 4
Shows the capsule and septa of the thymus gland. A part of the connective tissue of the capsule (arrowhead) invaginates the thymus to form the septae (arrow). The thymus appears surrounded by adipose tissue. H&E stain. Scale bar 50 µm.
Figure 5
Figure 5
Photomicrographs of a section in the thymus gland of a chick embryo at the 14th day of incubation. A: (control group) shows the lobules of the thymus consisting of epithelial cell mass (E) with a low number of lymphocytes surrounded by basement membrane (Red arrowhead) and the lobules separated by the interlobular mesenchyme(septa) (S). In the group treated with a high concentration of extract and in the mixed group, degradation and spacing can be seen between cells, as well as a reduction in cell density (Yellow arrow). (H&E) stain. Scale bar 50 µm.
Figure 6
Figure 6
Photomicrographs of a section in the thymus gland of chick embryo at 20th day of incubation. A: (control group) shows the lobules of the thymus, consisting of two regions, the cortex (C) and medulla (M), enclosed by a capsule (Red arrowhead); the lobules separated by septa (S) and the blood vessels also appear (black arrow). In the group treated with a high concentration of extracts and a mixed group, degradation and spacing could be seen between cells, as well as a reduction in cell density (Yellow arrow), and vacuoles (circle). (H&E) stain. Scale bar 50 µm.
Figure 7
Figure 7
Shows the level of IL-10 cytokine production that was determined in the serum of chick embryo using ELISA kits after embryo exposure to two doses of rosemary and thyme extracts in different concentrations with a dose of (0.1 mL). The data shown in the figure represent the mean ± SD (n = 4), and there is a significant difference between groups at a significance level of p < 0.05 *. At day 14, there are significant decreases in all groups exceptthe low-concentration thyme group. After three weeks of incubation and exposure to two doses of rosemary and thyme extract, a significant increase was recorded in all groups except the group treated with low-concentration rosemary.
Figure 8
Figure 8
Shows the level of IgM antibody production determined in the chick embryo serum using ELISA kits after embryo exposure to two doses of rosemary and thyme extracts in different concentrations, with a dose of 0.1 mL. The data shown in figure represent the mean ± SD (n = 4), and there is a significant difference between groups at the level of significance (p < 0.05) *. After two weeks of incubation, there was a significant decrease in high and medium concentrations of thyme and the mixed group, and a significant increase in the high-concentration rosemary group. At day 20, and after three weeks of incubation and exposure to two doses of rosemary and thyme extract, the IgM antibodies secretion was significantly decreased in all groups, except in the high-concentration thyme group, where there was an increase in antibody production.
Figure 9
Figure 9
Shows the level of IgG antibody production that was determined in the serum of chick embryo using ELISA kits after embryo exposure to two doses of rosemary and thyme extracts in different concentrations, and the dose was 0.1 mL. The data shown in the figure represent the mean ± SD (n = 4), and there is a significant difference between groups at the level of significance (p < 0.05) *. At day 14 and after two weeks of incubation, there was a significant decrease in all groups and a significant increase in high-concentration thyme. At day 20 and after three weeks of incubation and exposure to two doses of rosemary and thyme extract, there was a significant increase in antibody secretion in the group treated by high-concentration thyme and the mixed group, while a decrease was seen in all other groups except medium-concentration thyme.
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