تقييم الكفاءة الوقائية للمستخلص المائي لاوراق نبات Andrographis paniculata في بعض المعايير الفسلجية والنسجية والجزيئية في ذكور الارانب البيض المعاملة بمادة البوتيل هيدروكسي تولوين

Evaluation the Protective Efficiency of Aqueous Extract of Andrographis paniculata Leaves in Some Physiological , Histological and Molecular Parameters in White Male Rabbits Treated with Butylated Hydroxytoluene (BHT)

Summary
This study aims to know the protective role of the aqueous extract of Andrographis paniculata on some physiological and histological parameters in White male rabbits treated with butylated hydroxyltoluene (BHT).
The study was conducted in the College of Education for Pure Sciences / University of Karbala and the animal house of the College of Veterinary Medicine / University of Karbala for the period from the beginning of November 2021 until February 2022. This study included use of 100 male rabbits Oryctatagus cuniculus whose ages ranged between 8- 9 months with an average weight of 1500-2000 g. The aqueous extract of Andrographis paniculata was prepared, then the first experiment was conducted, which aimed to determine the half effective dose determining the half-effective dose (ED50) of the aqueous extract of Andrographis paniculata through study the curve of Effective dose, 60 adult male rabbits were used, which were randomly divided into six equal groups (10 animals/group) and they were administered orally and daily five increasing doses of Andrographis paniculata extract (0,50, 100, 150, 200 and 250 mg/day/ kg) for four weeks . Fasting blood samples were collected after the end of experiment to study the concentration of the following parameters: Malonaldehyde (MDA), Reduced Glutathione (GSH), Total Cholesterol (TC) and High-density Lipoprotein (HDL). The half-effect dose of was (100 mg/kg/B.W) .
The second experiment aimed to study the protective role of Andrographis paniculata extract on the toxic effects induced by BHT. 40 adult male rabbits were used, which were randomly divided into four equal groups (10 animals / group), the first group(G1) was administrated with 1 ml / kg of corn oil and considered as a control group, the second group (G2) administrated 1 mg / kg of BHT, the third group (G3) administrated 100 mg / kg of Andrographis , while the animals of the fourth group (G4) administrated 1 mg / kg of BHT and 100 mg / kg of Andrographis paniculata extract orally for a month daily.
Fasting Blood samples were collected after the end of the experiment to study the concentration of the following parameters: aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), superoxide dismutase (SOD), Malondialdehyde (MDA), Reduced Glutathion (GSH), High Density Lipoprotein (HDL), Low Density Lipoprotein (LDL), Very Low Density Lipoprotein (VLDL), Triacylglycerol (TAG), Total Bilirubin (T-BIL), Albumin, Urea, Uric acid, Tumor necrosis factor alpha (TNF-α), Interleukin 6 (IL- 6) and Cytochrome P450, in addition to measurement the gene expression of CD44 gene by using Real –Time PCR technology .
The results of this experiment showed that oral administration of BHT caused significant increase (P<0.01) in the concentrations of AST, ALP, ALT, MDA, TC, LDL, T-BIL, Urea, Uric acid, TNF-α, and IL-6, while there was significant decrease (P<0.01) in the concentration of Albumin, SOD, GSH, HDL and Cytochrome P450 , with no significant differences (P>0.01) in the concentration of TAG and VLDL compared with the control group.
The group that was treated with ED50 of Andrographis paniculata aqueous extract showed a significant increase (P<0.01) in the concentration of HDL, Albumin, GSH and Cytochrome P450 , while there was a significant decrease (P<0.01) in the concentration of TAG, LDL, VLDL, MDA, T-BIL and Urea. With no significant difference (P<0.01) in the concentrations of AST, ALP, ALT, TC, Uric acid, SOD, TNF-α and IL-6.
The group treated with ED50 of Andrographis paniculata aqueous extract showed a significant increase (P<0.01) in the concentration of HDL, Albumin and GSH , while there was a significant decrease (P<0.01) in the concentration of TAG, LDL, VLDL, MDA, T-BIL and Urea, With no significant difference (P>0.01) in the concentrations of AST, ALP, ALT, TC, Uric acid, SOD, TNF-α, IL-6 and Cytochrome P450 compared with the control group.
The experiment also showed that oral administration of BHT with oral administration of ED50 of andrographis aqueous extract, caused significant increase in GSH and no significant differences (P>0.01) in the concentration of ALP, AST, ALT, TC, TAG, HDL, LDL, VLDL, MDA, SOD, T-BIL, Albumin, Urea, Uric acid, TNF-α, IL-6 and Cytochrome P450 compared with the control group.
The results of the Molecular gene expression of CD44 gene showed significant increase in gene expression in the group treated with BHT when compared to the control group, while there was no significant increase in gene expression in the group treated with andrographis and BHT compared to the control group.
Histological examination showed that oral administration of BHT for months caused clear degenerative changes in the liver tissue with congestion in the central vein and infiltration of inflammatory cells around it, thickening, degeneration, rupture as well as hydrolysis of hepatocytes compared with the control group . In kidney tissue There was congestion and atrophy of the urinary glomerulus, in addition to increase in Bowman’s capsule space and destruction of the urinary tubules with the shedding of the inner lining of the urinary tubules. In the spleen there was degenerative changes , congestion and increase in the red pulp compared with control group.
It is concluded from this study that BHT causes clear pathological effects on the liver, kidneys and spleen, and confirms that treatment with the half effective dose of aqueous extract of andrographis (100 mg/kg) has a protective role against damage caused by BHT in white male rabbits.