Effects of Ocimum basilicum extract on hematological and serum profile of male albino mice after AlCl 3 induced toxicity

The present study was aimed to investigate the recovery effects of leave extract of Ocimum basilicum against AlCl3 induced toxicity. The trial was performed on adult male albino mice (Mus musculus). The experimental animals were randomly divided into three groups i.e. control, T1 and T2 with six animals in each group. Animals in control group received no treatment and were provided with normal saline. Both treatment groups T1 and T2 were orally administered with 80mg/kg body weight of AlCl3 for 16 days. After that animals in T1 and T2 groups were administered 100 mg/kg body weight and 200 mg/kg body weight respectively of Ocimum basilicum extract for 15 days. Several of the important hematological and serological parameters were measured. There was a significant increase in lymphocyte count and a significant decrease in neutrophil count in both T1 and T2 groups. There was a significant increase in red blood cells and platelet count while mean corpuscular haemoglobin was significantly decreased in T1 group. Alanine transaminase was significantly increased while serum creatinine was decreased in both T1 and T2 groups. It is concluded that Ocimum basilicum extract influenced several hematological and serological parameters in male albino adult mice after AlCl3 induced toxicity.


Introduction
There are several metal that have high concentration in the environment and are toxic for human beings as well as other living organisms [1,2].Aluminium (Al) is one of such metals frequently distributed through earth crust [3].It is an integral component of many medicines, food additives and also added to drinking water for purification purposes [4].Several sources of aluminium in daily life include spices, yellow cheese, aluminum ware, corn, salt, cosmetics, herbs, tea, antacid drugs and tooth paste [5].Aluminium accumulates in several mammalian tissues including liver, kidney, heart, bone, blood, and brain [6].The accumulated aluminium exerts toxic effects all over the human body especially central nervous system [7].It  It has also been used to cure several digestive disorders including nausea, poor digestion, abdominal cramps, gastro-enteritis, chronic diarrhea, dysentery; psychotic disorders like insomnia and depression as well as migraine [14].The present study aimed to evaluate the effects of the extract from Ocimum basilicum on recovery response in adult albino male mice after AlCl3 induced toxicity.

Experimental design
The experiment was conducted on male albino mice (aged approximately 8±1 weeks) obtained from the laboratory of Government Post Graduate College Gojra, Toba Tek Singh distrit, Pakistan.This experiment was carried out at animal house, Bio Park Bahaudin Zakaryia University.Mice were reared in small cages (locally manufactured) provided with wooden chips.Water and food were made available ad libitum.Room temperature (22 ± 1 0 C) was maintained along with a regular light/dark cycle in a ratio of 14:10.All the animal handling procedures and experimental protocols were approved by ethical review board of Institute of Pure and Applied Biology, Bahauddin Zakariya University Multan, Pakistan.The animals were divided randomly into 3 groups i.e. control, T1 and T2 (N = 6 in each group).The control group received normal saline, whereas experimental groups i.e.T1 and T2 were orally administered 80mg/kg body weight of AlCl3 for 16 days.After that, animals in T1 group were orally administered with 100 mg/kg body weight of Ocimum basilicum extract, and animals in T2 group were orally administered with 200mg/kg body weight of Ocimum basilicum extract for 15 days.

Leave extract of Ocimum basilicum
The leaves of Ocimum basilicum were collected from plants grown in local fields of Multan.The leaves were made free of adulterated material through manual picking, washed and shade dried.Dried leaves were powdered with the help of Waring Blender.300 g of powdered leaves was soaked in 70 % aqueous ethanol (v/v) and 30 % distilled water for 7 days in amber colored glass bottles.The bottles were shaken at least 3-4 times per day.The gas was removed by opening lid of the bottle after every shake and then was tied tightly again.The soaked leaves material was filtered first using a double layered muslin cloth to remove vegetative debris, and finally through Whatman filter paper (No. 4).The residues were re-soaked in 70 % aqueous-ethanol and 30 % distilled water and the procedure was repeated twice.The obtained crystal clear filtrates were evaporated on a rotatory evaporator (R-210, BUCHI, Switzerland) under reduced pressure (-760 mmHg) and temperature (37 o C) to a concentrated paste.The thick pastes were kept in their respective petri plates and stored at 4 o C until further use.

Statistical analysis
All the data is expressed as means ±standard deviation (SD).The data obtained in this analysis was subjected to two sample t-test for comparing hematological and serum parameters of control and treatment groups, using statistical package Minitab (version 17, Pennsylvania).

Hematological parameters
In T1 group, RBC and PLT and LYM showed significant increase compared to control group; while MCH and NEUT showed significant decrease compared to control group.The WBC, HGB, HCT, MCV and MCHC showed no significant changes in T1 group compared to control group.In T2 group, LYM showed significant increase while NEUT showed significant decrease compared to control group.There was no significant change in RBC, WBC, HGB, HCT, MCV, MCH, MCHC, and PLT compared to control group (Table 1).

Serological parameters
In T1 group, AST showed significant increase; while cholesterol and creatinine showed significant decrease compared to control group.Whereas, ALT, triglyceride and total protein showed no significant change compared to control group.In T2 group, AST showed significant increase compared to control group; while creatinine showed significant decrease compared to control group.ALT, cholesterol, triglyceride and total protein showed no significant change compared to control group (Table 2).

Table 2 . Effect of administration of Ocimum basilicum extract on blood serum profile of albino male mice for recovery response after AlCl3 induced toxicity
This difference might be due to difference in pretreatment as we used AlCl3 in our experiment while paracetamol was used as pretreatment by Khoun [22].Our results also demonstrated that ethanolic leaf extract of Ocimum basilicum significantly decreased serum creatinine level.These findings are in agreement with the previous results which demonstrate that aqueous extracts of Ocimum gratissimum in wistar albino rats [27], Ocimum basilicum [28], and ocimum sanctum in mice [21] significantly decreased serum creatinine level.This decrease in serum creatinine level might be due to nephroprotective effects of Ocimum.Conclusions Leave extract of Ocimum basilicum influenced several hematological parameters including red blood cell count, mean corpuscular hemoglobin, lymphocyte and neutrophil count in male albino adult mice after AlCl3 induced toxicity.Similarly, several serological parameters including aspartate transaminase, cholesterol and creatinine were also influenced by oral administration of the leave extract of Ocimum basilicum.