دراسة التأثير الوقائي للمستخلص الكحولي والنانوي لبذور نبات الخرفيش Silybum marianum على بعض المعايير الفسلجية والنسجية والوراثية في ذكور الجرذان المستحث بها سرطان الكبد

Study of the protective effect of alcoholic and nano-extract of Silybum marianum seeds on some physiological, histological and genetic parameters

The current study aimed to know the protective effect of alcoholic and nano-synthesized zinc oxide extract of Silybum marianum seeds to reduce the effect of liver cancer induced by thioacetamide (TAA) and carbon tetrachloride (CCl4) in male white rats, Rattus Rattus, by studying some changes in parameters. Physiological, histological and genetic. The study was conducted in the animal house of the College of Education for Pure Sciences – University of Karbala for the period from 1/3/2022 to 1/7/2022, it included 35 male white rats with weights (200-250) grams and their ages were (10-12) weeks. It was divided into seven groups of 5 animals per group. The first group, G1, was a negative control group that was dosed with physiological salt solution for a period of 14 weeks, and the second group, G2, was dosed with an alcoholic extract of plant seeds at a dosage of 100mg/kg for a period of 14 weeks. The third group, G3, was a positive control that was injected intraperitoneally with thioacetamide at a dose 200mg/kg daily for two weeks, then injected with carbon tetrachloride at a dose of 3mg/kg once a week for 12 weeks, and the fourth group, G4, was dosed with alcoholic extract at a dose of 100 mg/kg for two weeks, after which the mixture of thioacetamide and carbon tetrachloride was given by injection until the end of the experiment. It is 14 weeks, while the fifth group (G5), the sixth (G6) and the seventh (G7) were dosed with the nano-extract (20,32,60 mg/kg), respectively, for two weeks, and then injected with a mixture of thioacetamide and carbon tetrachloride until the end of the experiment, which is 14 weeks. Blood and tissue samples were collected after sacrificing the animals at the end of the experiment to measure the following parameters : Measuring the level of liver enzymes Alanine transaminase (ALT), Aspartate transaminase (AST), Alkaline phosphatase (ALP), Total protein, Albumin, Globulin, Malondialdehyde, Glutathione, and Superoxide dismutase , as well as evaluating the pathological changes of the liver tissue by measuring the diameters of each of the hepatocytes, sinusoids, and central vein, and measuring the percentage of DNA damage. We obtained the following results:
1- The study revealed the presence of 12 active compounds present in the alcoholic extract of artichoke seeds by using GC-Mass technology, and the active compound was Phenol, 2,2′-methylenebis[6-(1,1-dimethylethyl)-4-methyl- has The highest surface area in the extract, while the active compound Hexadecen-1-ol,trans-9- had the lowest surface area in the extract.
2- There was a significant increase (p≤0.05) in the level of liver enzymes AST, ALT, ALP and malonaldehyde in the positive control group G3 compared with the negative control group G1, while there were no significant differences (p≥0.05) between G1 and G2, while there was a decrease Significant (p≤0.05) in the level of liver enzymes AST, ALT, and ALP in male rats in group G4 that were dosed with alcoholic extract of artichoke seeds, as well as in groups G7, G6, and G5 that were dosed with nanoextract, as well as a significant decrease (p≤0.05). In the level of total protein, albumin, globulin, cotathione, and superoxide dismutase in the positive control group G3 compared with the negative control group G1, while there were no significant differences (p≥0.05) between G1 and G2 except for the level of glutathione, there was a significant difference (p≤0.05) between G1 and G2. While there was a significant increase (p≤0.05) in the level of total protein, albumin, globulin, cotathione, and superoxide dismutase in male rats in the G4 group that were dosed with the alcoholic extract of the milk thistle plant, as well as in the groups G7, G6, and G5 that were dosed with the nano extract of the plant.
3- There were changes in the liver tissue of the positive control group G3 compared to the negative control group G1 and group G2, and it included the presence of tumor cells and necrosis in the liver cells with hemorrhage in the tissue and irregular sinusoids as well as irregular shape of the hepatic lobule with inflammation in some areas of the tissue, while the group G4 Histopathology shows degeneration in some hepatocytes and necrosis in other cells with irregular sinusoids. As for the groups G7, G6, and G5 that were dosed with the nanoextract, they showed simple histological changes compared to the positive control group represented by degeneration in some liver cells in G5 and mild inflammation in G6. The histological sections are regular in the sinusoids, and the shape of the tissue was normal.
As well as the presence of a significant increase (p≤0.05) in the diameters of liver cells, sinusoids and central vein in the positive control group G3 compared with the negative control group G1, while there were no significant differences (p≥0.05) between G1 and G2, while there was a significant decrease (p≤ (0.05) in the diameters of liver cells and the central vein in male rats in group G4 that were dosed with alcoholic extract of the milk thistle plant, and there was a significant decrease (p≤0.05) in the diameters of liver cells, sinusoids and central vein in male rats in groups G7, G6, and G5 that were dosed with the extract. nanoparticles.
4- There was a significant increase (p≤0.05) in the percentage of DNA breakdown in the positive control group G3 compared to the negative control group G1, while there were no significant differences (p≥0.05) between G1 and G2, while there was a significant decrease (p≤0.05). In the percentage of DNA breakdown in male rats in group G4 that were dosed with alcoholic extract of artichoke seeds, as well as in groups G7, G6, and G5 that were dosed with nanoextract, compared to the positive control group, G3.
We conclude from the foregoing that the induction of liver cancer in male white rats by injecting them subperitoneally with TAA and CCl4 leads to changes in some physiological, histological and genetic parameters when the animals were dosed orally with the alcoholic and nano extract of the seeds of the Artemisia plant. It had a protective effect against liver cancer and the nano extract with Higher protective efficacy than the alcoholic extract of the plant’s seeds, especially at the dose (32mg/kg).