Humans are inexorably out to metals attributable to their ubiquity in nature, polluted air, water, soil and food, wide use in industry and long-term persistence in the environment. Metals may have serious effect on the male reproductive system directly, when they target specifi reproductive organs, or indirectly, when they act on the neuroendocrine system. Metals have been shown to affect spermatogenesis in rodents and humans, which can lead to low sperm count, abnormal sperm morphology and poor semen quality [1-2]. Among them aluminum is the most widely distributed trivalent cation found in its ionic form in most kinds of animal and plant tissues and in natural water everywhere.

It is the third most prevalent element and the most abundant metal in the earth's crust, representing approximately 8% of total mineral components [3]. Aluminium occurs naturally in the environment and is also released due to anthropogenic activities such as mining and industrial uses, in the production of aluminium metal and other forms of aluminium compounds. A variety of aluminium compounds are produced and used for different purposes, such as in water treatment, papermaking, fire retardant, fillers, food additives, colors and pharmaceuticals. Previously, Aluminium has been considered on an indifferent element from a toxicological point of view for a long time. Although aluminium is present in trace amounts in the biological material, it does not appear to be an essential element and is usually considered to have harmful effects on general health [3]. Aluminium is known as a neurotoxin that can cause certain diseases such as Alzheimer disease, dialysis dementia, Parkinsonism, and amyotropic lateral sclerosis. In addition, [4] to its neurotoxicity, aluminium affects other body structures like the skeletal system [5], brain tissue, bone, blood cells, liver and kidney [6]. The sources of aluminium are specially corn, yellow cheese, salt, herbs, spices, tea, cosmetics, aluminium ware and containers. Also, aluminium is widely used in antacid drugs, as well as in food additives and toothpaste [1]. Environmental pollution with the different aluminium containing compounds, especially those in industrial waste water, exposes people to higher than normal levels of aluminium. This experiment was carried out to determine the effect of aqueous extract of Massularia acuminata on the hematological parameters of rats treated with aluminium chloride. The kidney is an essential organ required by the body to perform several important functions including the maintenance of homeostasis, regulation of the extracellular environment, such as detoxification, and excretion of toxic metabolites and drugs [7]. The functional integrity of the mammalian kidney is vital to total body homeostasis because the kidney plays a principal role in the excretion of metabolic wastes and in the regulation of extracellular fluid volume, electrolyte composition, and acid–base balance. In addition, the kidney synthesizes and releases hormones, such as renin and erythropoietin, and metabolizes vitamin D3 to the active 1,25-dihydroxyvitamin D3 form. A toxic insult to the kidney therefore could disrupt any or all of these functions and could have profound effects on total body metabolism. The mammalian kidney, because of its high rate of perfusion, active transport capabilities, and concentrating functions, often is exposed to much higher concentrations of chemicals than are other organs. These high concentrations of chemicals may have no effect on the kidney, may produce a beneficial therapeutic response, or may produce harmful (toxic) effects to the organ. Therefore, the kidney can be considered as a major target organ for exogenous toxicants. Nephrotoxicity is a kidney-specific feature in which excretion does not go smoothly owing to toxic chemicals or drugs [8]. Approximately 20% of nephrotoxocity is induced by drugs, but medication of the elderly increases the incidence of nephrotoxicity up to 66% as the average life span increases. Chemotherapy or anticancer medicine has been of limited use due to nephrotoxicity [9]. Some chemicals may produce a direct renal cytotoxicity, such as that produced by heavy metals (mercury, chromium), by therapeutic agents (aminoglycoside antibiotics, analgesics) or by a variety of widely used chemicals (chloroform, carbon tetrachloride). Certain other chemicals in the environment may produce biochemical changes in the kidney that alone appear to have no functional correlation (that is, no functional nephropathy can be measured) yet may alter the response of the kidney to other agents. The former chemicals produce identifiable functional lesions that may be life-threatening. It has been reported previously that there is a negative correlation between endogenous testosterone levels and kidney disease stages [10].The mechanism for this is likely to involve some alteration or derangement of the male reproductive hormone profile [11]. This supports the current findings of a significant decrease in testosterone levels in kidney disease stages , a significant increase in the leuteinizing hormone (LH) level and a pattern of hypergonadotropic hypogonadism were seen. This suggests that uremic metabolites secondary to advanced kidney disease stage affect the testes more than the hypothalamic or pituitary function. Alternatively,the degradation of uremic metabolites in the hypothalamic or pituitary region is faster and more pronounced in the testes. An alternative,more trival, explanation is that LH and prolactin are polypeptide protein hormones (whereas testosterone is not) whose frenal clearance may be impairedas GFR decreases and, hence,the observed elevated levels oh LH and prolactin are merely artifactual. Massularia acuminata (Rubiaceae) known as pako ijebu or orin ijebu (Yoruba- Western Nigeria), is a tree growing up to 5 m high. It is distributed from Sierria Leone through Nigeria to Democratic Republic of Congo. The large leaves are practically stalkless, elliptic, acuminate and almost glaborious. [12]. The juice from the fruit is used as antibiotics for the treatment of eye infections in Sierra Leone. The stems are used as chewing stick for oral hygeine in Nigeria [13]. The decoction or infusion of the stem has also been claimed to be used as aphrodisiac and anticarcinogen [14]. Previous studies have reported that the alkaloidal content of Massularia acuminata stem was responsible for the antibacterial and anti-inflammatory activities of the plant [15]. Futhermore, a recent study have also validated the aphrodisiac claim of aqueous extract of Massularia acuminata root in male rats. In addition, [16] it was reported that the aqueous extract of Massularia acuminata stem exhibited dose related androgenic and gonadotropic effects in male rats. Thus, it is logical to investigate the acclaimed aphrodisiac potential of the plant stem in a complete randomized design at the same doses used previously by [16]. The present investigation was therefore undertaken to evaluate the aphrodisiac activity and therapeutic effects of aqueous extract of Massularia acuminata stem using doses (100, 300 and 500 mg/kg body weight) which produced significant and dose related androgenic effects in male rats. This is with a view to validating the acclaimed use of the plant stem as sexual invigorator and other therapeutic use in the folk medicine of Nigeria.

Aluminium Chloride

Aluminium is the third most abundant element on the earth crust and its compounds are widely distributed in nature. It is used in the manufacturing of many everyday products like toothpastes, antiperspirants, cosmetics, processed foods, adjuvants in various parenteral preparations and pharmaceutical agents [17-18]. This metal is incorporated in some medications such as antacids, buffered aspirins and anti-diarrheal products. Due to its reactivity, aluminum in nature is found only in combination with other elements such as sulfate, chloride etc. Aluminium sulfate is extensively added as a coagulant agent during the purification process of drinking water in order to folliculate the organic matter, to clarify the water [19]. Aluminium is a metal of choice in making various kinds of household cookware and storage utensils. Aluminium can enter the body through inhalation of air contaminated with aluminium compounds, through ingestion of aluminium dusts or with food and drinking water and through dermal contact. In the body, aluminium accumulates mainly in bones, liver, testes, kidney and brain [3]. Different forms of aluminium are environmental xenobiotics that induce free radical-mediated cytotoxicity [20]. Aluminium accelerates oxidative damage to lipids, proteins and nucleic acids. Different studies demonstrated that the main toxic effects of aluminium in a chronic exposure were neurological with encephalopathy and psychomotor functions disturbances, in bone with osteomalacia and hematological with microcytic anemia [21]. An experiment reported that aluminium induced toxicity in epididymis, vas deferens, seminal vesicle and ventral prostate in mice [22]. According to another study, it has been shown that aluminium chloride induced reproductive toxicity and exerted a significant effect on the steroidogenesis [23]. It has been shown in a study that, in vitro, aluminium chloride provoked deterioration in sperm motility and viability and enhancement of free radicals and alterations in enzyme activities on rabbit sperm [24]. Alterations in the metabolism of testis and epididymis, leading to a reduction in fertility rate in mice treated with aluminium chloride were also observed in an Individual study [20]. There have been report from epidemiological evaluation that infertility due to male factor ranged from 20% to 70% and infertility rates were highest in Africa and Eastern Europe [25]. There are evidences to show that sperm counts have been declining over the last 50 years with a consequent increase in male infertility [26].

Massularia acuminata

Massularia acuminata (G. Don) Bullock ex Hoyl. (Rubiaceae) known as pako ijebu or orin ijebu (Yoruba- Western Nigeria), is a tree growing up to 5 m high. It is distributed from Sierria Leone through Nigeria to Democratic Republic of Congo. The large leaves are practically stalkless, elliptic, acuminate and almost glaborious. The juice from the fruit is used as antibiotics for the treatment of eye infections in Sierra Leone. The stems are used as chewing stick for oral hygiene in Nigeria [13]. The decoction or infusion of the stem has also been claimed to be used as aphrodisiac and anticarcinogen [14]. Previous studies have reported that the alkaloidal content of M. acuminata stem was responsible for the antibacterial and anti-inflammatory activities of the plant [27]. Futhermore, a recent study by Yakubu et al. [24] have also validated the aphrodisiac claim of aqueous extract of M. acuminata root at 50, 100, and 200 mg/kg body weight in male rats. In addition, Yakubu et al. [28] have also reported that the aqueous extract of M. acuminata stem at the doses of 250, 500, and 1000 mg/kg body weight exhibited dose related androgenic and gonadotropic effects in male rats. Thus, it is logical to investigate the acclaimed aphrodisiac potential of the plant stem in a complete randomized design at the same doses used previously by Yakubu et al. [28].

Collection and Extraction of Plant Materials

Plant of Massularia acuminata was collected in at Owo in Ondo State, Nigeria. The plant was authenticated at Adekunle Ajasin University, Akungba Akoko, Ondo state. Fresh bark of Massularia acuminata were some ted out and washed to remove debris and dust particles. The extract was kept in the refrigerator.

Treatment of Experimental Animals

The animals used were 35 healthy white male albino rats, 2–2.5 months old weighing between 140–176 g were obtained from the university college Hospital Ibadan, Oyo state, Nigeria. The animals were housed in clean aluminum cages placed in well-ventilated house conditions (Temperature: 28–31°C; photoperiod: 12 h natural light and 12 h dark; humidity: 50–55 %). They were also allowed unrestricted access to rat pellets (Bendel Feeds and Flour Mills Ltd., Ewu, Nigeria) and tap water. The animals were housed in standard cages and kept at room temperature. Approval for study was obtained from the department of Biochemistry, Adekunle Ajasin University, Akungba Akoko, Ondo State. All experiments involving animals were conducted at the animal house of Adekunle Ajasin University, Akungba Akoko, Ondo State.

Aluminium Chloride

Three cages filled with 6 albino rats each were dosed with aluminium chloride at different concentration (100 mg/kg body weight, 300 mg/kg body weight,500 mg/kg body weight respectively) and were isolated respectively in their different cages ( Figure 1).

Figure 1: Formulas for Calculating Hematological Parameters

Experimental Design

Thirty-five male albino rats were used and they were divided into seven groups, each consisting of five animals each.

• Group 1: Basal control: the animals were niether given Aluminium Chloride (Toxicant) nor Massularia acuminata (Extract)

• Group 2: Negative control (N – CTR): the animals in this group were orally administered with the aluminium Chloride (Toxicant)

• Group 3: Positive control (P – CTR): the animals in this group were administered with Sildenafil Citrate only

• Group 4: The animals in this group were administered with both Aluminium chloride and Sildenafil Citrate

• Group 5: The animals in this group were administered with both Aluminium chloride and 100 mg/kg M. acuminata

• Group 6: The animals in this group were administered with both Aluminium chloride and 300 mg/kg M. acuminata

• Group 7: The animals in this group were administered with both Aluminium chloride and 500 mg/kg M. acuminata

At the end of the 24^th day period of stable administration, animals were subjected to fasting over- night and later sacrificed in the next morning. Blood samples were collected directly from the orbital sinus of the dissected animals using a hematocrit capillary and then poured into an EDTA bottle and taken to the laboratory for the hematological test. Tissues which include liver, kidney, brain, testes and heart were excised from all the subjects (animals) into well labeled tubes containing normal saline across the groups. Tissues were then stored under the temperature of -7⁰c for 24 hours.

Biochemical Parameters

Kidney function test was done by determining the level of urea and electrolytes in the serum of all the groups studied.

Determination of Serum Total Protein Activity

The concentration of serum total protein was determined using the method of Tietz as outlined in Randox kits, UK.

Determination of Aspartate Aminotransferase (AST) Activity

The activity of aspartate amino transferase was assayed by the methods of Reitman and Frankel as outlined in Randox Kit.

Determination of Alanine Aminotransferase (ALT) Activity

The activity of alanine amino transferase was assayed by the methods of Reitman and Frankel as outlined in Randox Kits, UK.

Determination of Alkaline Phosphatase (ALP) Activity

The activity of alkaline phosphatase (ALP) was assayed by the method of lkein et al. as outlined in Randox Kits, UK.

Determination of Haematological Parameters

Pooled blood was collected from the animal per treatment by capillary tubes through the ocular punctures. The collected blood was placed in coded 1.5mL heparinized plastic tubes, stored on ice according to the procedures established by Campbell and Murru. Haematocrit (Hct; %) or Packed cell volume (PCV) in the blood samples was assayed by microcentrifugation at 3500 × g for 10 minutes of standard heparinized microhaematocrit capillary tubes and measuring the percentage of packed cell volume [17]. Haemoglobin concentration (Hb; g dL^-1) was spectrophotometrically measured on the basis of cyanmethemoglobin procedure by Drabkin. The number of red (RBC) and white blood cell (WBC) were counted microscopically in a haemocytometer, using an improved Neubauer chamber after diluting blood samples with Hayem solution (for RBC) or Turk solution (for WBC). Mean corpuscular volume (MCV; fl), mean corpuscular haemoglobin (MCHb; pg) and mean corpuscular haemoglobin concentration (MCHC %); were calculated for each sample according to the method of Klinger et al. as shown.

The effects of aqueous Massularia acuminata leave extract on serum biochemical parameters of experimental rats is shown in Tables 1. 

Table 1: Effects of Treatment of Massularia acuminata Stem Bark on Liver and Kidney Functions Enzymes (Enzymes and Non-enzymes)

The results indicated that levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and urea were found to be higher (p< 0.05) in the toxicant treated groups of the experimental animals, particularly at the dose (35 mg/kg body weight) compared to the control, whereas serum concentrations of total protein, albumin and bilirubin were unaltered in the animals in the first phase of the study. Aspartate aminotransferase (AST) in heart, liver and kidney was significantly (p<0.05) reduced in group treated with different doses of aqueous extracts of Massularia acuminata compared to control group. There was an elevation of alanine aminotransferase (ALT) in all treated group but not significantly (p>0.05) different from the control group, while group treated with aqueous extract showed a reduced ALT when compared with the control and it is statistically significant (p<0.05). Alkaline phosphatase in the liver, serum and testis (Table 1) was markedly decreased in the control group but the groups treated with the toxicant and the extracts has a slightly higher ALP level and it is significant (p<0.05) when compared to the control but the control group was significantly (p<0.05) different from the control group. The result of haematological parameters is shown in Table 2. 

Table 2: Effects of Treatment of Massularia acuminata Stem Bark on Haematological Parameters

The results of effects of Massularia acuminata on haematological parameters in rats are presented in Tables 2. White blood cell count was significantly steadily increase (p<0.05) in the groups that received Massularia acuminata as compared to the control group in the rats. All other Haematological parameters were similar in all the groups. The effects of the Massularia acuminata on platelets, PCV and haemoglobin parameters in rats treated with Massularia acuminata was significantly (p<0.05) higher across all the intervention groups than the control. There was a sharp increase in Mean corpuscular volume (MCV) concentration in all the treated groups. Similarly, the percentage Mean corpuscular haemoglobin (MCH) showed a dose- dependent elevation in all the treated groups and significantly (p<0.05) different from the control group. WBC, was significantly (p<0.05) higher in treated groups; similarly, RBC was significantly (p<0.05) elevated in the same groups when compared with the control group. There was also a significant (p<0.05) increase in HGB in all the treated groups when compared with the control group. HCT and LYM was also statistically (p<0.05) higher in all the treated groups as compared to the control group. The histology results of the liver shown in Plate 1 and 2 respectively revealed a clear and normal histology of the architecture of the hepatocytes across all the group with clear sinusoid and central vein (Figree 2).

Figure 2: Histopathological Examination of Liver Tissues

Group 1 = Control; Group 2 = AlCl₃; Group 3 = Sildenafil Citrate; Group 4 = Toxicant+Sildenafil Citrate; Group 5 = AlCl₃+100 Mg/kg M. acuminata; Group 6 = AlCl₃+300 Mg/kg M. acuminata; Group 7 = AlCl3 + 500 Mg/kg M. acuminata. Values with the letter (a) are statistically higher than the control; while values with the letter (b) are statistically lower than the control. Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Alkaline phosphatase (ALP), Total protein (TP).

Group 1 = Control; Group 2 = AlCl₃; Group 3 = Sildenafil Citrate; Group 4 =Toxicant+Sildenafil Citrate; Group 5 = AlCl₃+100 Mg/kg M. acuminata; Group 6 = AlCl₃+300 Mg/kg M. acuminata; Group 7 = AlCl3 + 500 Mg/kg M. acuminata.

Exposure to hazardous or toxic substances can affect the body in many ways. In general, when chemicals and other hazardous substances are absorbed, they travel through the various body systems and can affect a particular organ or organs, called the target organ(s). Fortunately, the body has mechanisms, mainly in the liver and kidneys, to process and eliminate many of these substances. This ability to eliminate toxic substances can reduce the effect(s) on the target organ(s). The Amino transferases (ALT and AST) are biochemical parameters and biomarkers useful for evaluating the functional integrity of organs. Amino transferases (ALT and AST) are sensitive indicators of hepatocellular damage. Alanine amino transferases (ALT) is widely distributed in liver, kidney, pancreas, lungs, brain, cardiac and skeletal muscles, whereas aspartate amino tranferases (AST) is mainly found in the liver. Alanine amino transferases is cytosolic while aspartate amino transferases are found in both cytoplasm and mitochondria of hepatic cells. The results of liver and kidney functional enzymes in Table 1, shows a significant elevated levels in the liver and kidney function enzymes in the aluminium chloride- treated groups when compared to the control which perhaps may indicates the toxicity of aluminium chloride to the liver. Dose-related elevated levels of these liver enzymes across all the group observed in this study (Tables 1) indicate some degree of hepatocellular damage induced by the aluminium chloride. There are significant elevated levels in the results of AST in serum, liver and kidney in the group treated with Massularia acuminata when compared to the control which indicates the dose of extracts of aluminum chloride administered probably imposed some degree of hepatocellular damages to the animals, but when the results is compared to the the toxicant treated group, there was a decreased in the level of this enzymes which confers a protective and ameliorative abilities of the plant extract to salvage the organ from biochemical damage. The prolonged administration of the extract might have caused these changes in serum enzymes. However, elevation of enzymes may not reflect generalized systemic toxicity. Although levels of aminotransferases indicate hepatocellular damage, they do not solely reflect the severity of the damage. Moreso, the results of the ALT in the serum and kidney equally reveal a significant elevated level of these enzymes in toxicant-treated groups which perhaps may indicate a hepatocellular damage, whereas, there is a significant low level of these enzymes in extracts- treated groups at different doses which show some protective and recovery effect of the extract on the organs. Elevated urea concentrations in liver of AlCl₃- exposed rats showed nephrotoxicity (Table 1). This elevation in urea might be due to damage produced in kidney tubules and this was confirmed by marked alterations in renal tissues when compared to the control group the administration of AlCl₃ exerts possible hepatotoxicity as verified by a significant increase in serum ALT, AST activities level (Table 1). In fact, these enzymes are known as important markers of hepatocellular damage as affirmed by Abdel-Moneim et al. This damage was in accordance with the cellular damages and loss of hepatic tissue structural pattern in AlCl₃-treated animals. The elevation in liver enzymes, direct and total bilirubin level has been well supported by Sheikh et al. and Nwokocha et al.who have demonstrated that aluminium causes hepatotoxicity. At the same time, Massularia acuminata remarkably inhibited aluminium-induced liver damage as evidenced by significant decreased activities of serum AST, ALT levels in the liver, serum and kidney of the experimental animals and restored to nearly normal architecture of the hepatocytes These results were in agreement with Sindhu et al. who reported that the reversal of increased serum enzymes in acetaminophen induced liver damage by berberine. Alkaline phosphatase (ALP) is predominant in the microvillii of the bile canaliculi and is used to assess the integrity of plasma membrane. An elevated serum level of ALP in the toxicant group is an established finding of cholestatic liver disease. The increase in ALP without a concomitant rise in bilirubin observed in this study might suggest some mild form of cholestatic disease. Serum total protein, albumin and bilirubin concentrations indicate both the state and severity of hepatic injury. The normal levels of protein, albumin and bilirubin observed in the present study (Tables 1) suggest that the synthetic and secretory functions of the liver were not impaired by the extract and may also indicate that the extract is not completely hepatotoxic. Albumin is the protein with the highest concentration in the plasma. It transports many substances including drugs and prevents fluid leakage into the tissues. Elevated level of bilirubin, a product of haem component of haemoglobin is associated with haemolytic anaemia, biliary obstruction or hepatic disease. The levels of bilirubin determined in this study were unaffected by extract of Massularia acuminata suggesting that there was no treatment related destruction of erythrocytes by the plant extract. The results might also indicate that the capacity of hepatic cells to excrete bilirubin was not adversely affected by the extract. Urea, a non-protein nitrogenous substance is a by- product of protein catabolism and formed in the liver, but excreted by the kidney. Elevated levels of urea in blood can be attributed to high protein diet, dehydration, severe haemorrhage and shock. The increased level of urea observed in this study (Table 1) especially at high dosage levels is suggestive of an impaired renal capacity to excrete the waste product.

However, the histopathological findings did not indicate nephrotoxicity as reflected by the preserved renal corpuscles. The protective effect of flavonoid and tannin against nephrotoxicity has been reported. From the results obtained in the study; aluminium chloride caused rise in alanine aminotransferases (ALT), urea, AST, in the liver, serum and kidney; and is significant (p>0.05) when compared to control. It is an indication of hepatocellular injury. ALT is a cytoplasmic enzyme found in very high concentration in the liver (Aliyu et al., 2007), and an increase of this specific enzyme indicates hepatocellular damage, while AST is less specific than ALT as an indicator of liver function.

The toxicity by aluminium chloride on the hepatocytes was also confirmed from histological sectioning which indicated various degrees of spotty hepatocellular necrosis in all the toxicant groups. The observed damage may be due to the fact that the liver being the first target of acute toxicity and the first organ exposed to everything that is absorbed in the small intestine, may metabolize foreign substances to highly reactive metabolites which may be hepatotoxic. In addition, because of the short duration of treatment, the alterations might be incipient and reversible, and not pronounced enough to change significantly serum ALT levels. The decreased levels of ALT, and AST in all the various extracts of Massularia acuminata groups in a dose-dependent manner, probably confirm the restorative ability of the extract but the result and is significant (p<0.05) when compared with the control and toxicant groups. These results were in agreement with Sindhu et al. who reported that the reversal of increased serum enzymes in acetaminophen induced liver damage by berberine. Furthermore, rats pre-treated with extracts of Massularia acuminata at different doses as well as the crude extract showed significant decreased serum AST activities, total and direct bilirubin level when compared with the AlCl3-exposed rats at p<0.05. The estimation of total proteins in the body is helpful in differentiating between a normal and damaged liver function as the majority of plasma proteins like albumins and globulins are produced in the liver. It is the main protein in blood and is made by the liver. It is clear from this study that an administration of aluminum chloride in rats led to reduction of protein concentration in serum of the animals compared to control. But an appreciable high level of protein was seen with treatment with fractions of the Massularia acuminata which invariably may boast the normal functioning of the animal in relation to protein functions. Thus the extract of Massularia acuminata has ability to restore protein synthesis and thus the normal functions in the animal.

Analysis of blood parameters is relevant to risk evaluation as the changes in the hematological system have a higher predictive value for human toxicity, when the data is translated from animal studies. Sub-acute exposure of rats to both lower and higher doses of the extracts of Massularia acuminata, produced significant changes in some haematological parameters in both studies. The decrease in haematological parameters like RBC, WBC, PLT, PCV, MCV, and RDW (Table 2) in the toxicant-treated groups revealed a detrimental effects of the toxicant to the animals in that groups and it is significant when compared to both the control and extracts-treated intervention groups. The increase or elevated level in erythrocytes (RBC), mean corpuscular haemaglobin concentration (MCV), Neutrophil, RDW, MCH leukocytes (WBC) and platetcrit (PCT) in the extract- treated groups and sildenafil-treated group in Table 2 which is not significant suggest that feeding the experimental animals with extracts of Massularia acuminata may not affect or compromise the health status of the animals. The elevated values may probably due to the choice of the sex of the animals selected for the study, as male animals have been found to have elevated hematological values when compared to their female counterparts, gender have been implicated as one of the most important factor influencing the haematological parameters of experimental animals. RBC counts have been found to be higher in male animals than the female folds. The high value in haematological parameters may probably be due to overproduction of hematopoietic regulatory elements such as colony-stimulating factors, erythropoietin, and thrombopoietin by the stromal cells and macrophages in the bone marrow thus providing the local environment for hematopoiesis. WBC and RDW were also found to be significantly elevated across all the groups treated with extract of Massularia acuminata. From the observed value of WBC in extracts of Massularia acuminata, it is clear that an increase in the number of WBC is a normal reaction of rats to foreign substances, which alter their normal physiological processes. The leukocytosis observed in this study indicates a stimulation of the immune system which protects the rats against infection and oxidative stress that might have been caused by reactive oxygen species generated by aluminum chloride and other secondary infections, which may be contracted after the weakening condition of the rats. Leukocytosis, which may be directly proportional to the severity of the causative stress condition, may be attributed to an increase in leukocyte mobilization. The elevated level of MPV and PDW in the extract may be due to high rate of erythropoiesis occurring in the bone marrow of the rats. In the second phase of the study, there were significant (p<0.05) reduction in the level of RBC, PCV, WBC, LYM and HCT in aluminium chloride treated groups when compared to the control. All the haematological parameters were found to be significantly elevated in extract of Massularia acuminata stem bark treated groups and its significant when compared to the control and toxicant-treated groups. The extracts did not alter the haematological parameters of the animals (Tables 2), an indication that there was no interference on red blood cell production. Red blood cells (RBC) are important in transporting respiratory gases. That there was no treatment related effects on RBC and haemoglobin (Hb) implies that the extract did not adversely affect the oxygen carrying capacity of the blood and the amount of oxygen delivered to the tissues. Erythrocytic indices (MCV, MCH and MCHC) are important in diagnosing anaemia. Some medicinal plants are known to cause destruction of red blood cells leading to anaemia. Massularia acumuinata stem bark appears to have no such potential of inducing anaemia, as it boasts the normal functioning of the haematological parameters, viz-a-viz the RBC. The observed increase in total WBC count indicates an enhanced phagocytic function of the leucocytes. Platelets play important role in the process of homeostasis and its reduction might adversely affect thrombopoietin. The increased platelet count following oral administration of Massularia acuminata extract indicates that the extract may not cause any coagulation problem, but has the potential of enhancing clotting and preventing haemorrhages. In contrast, anticoagulant property of Massularia acuminata was inferred because the extract precipitated low level of platelets in circulation.

The result of the histology of the photomicrograph of the liver revealed a normal of the hepnhatocyte is observed across all the groups with clear central veins, which indicates that the extract of Massularia acuminata preserves the normal architecture of the hepatocytes and thus the cell functions. However, these results showed Massularia acuminata has positive therapeutic effect on the kidney, it has the ability to treat inlammation and neurosis of the kidney and kidney diseases due to reactive oxygen species, thus administration of aqueous extract of Massularia acuminata at lower doses may be useful in alleviating and salvaging kidney and liver toxicities.

In conclusion, from the results obtained above, it could be inferred that the aqueous leave extracts of Massularia acuminata possess a great potential of ameliorating and restoring hepatocyte damage induced by any toxic chemicals and the potency of the extract, also may also find a great influence in boasting the immune system and hematological parameters in the animals which are the keys to survival of the cell to normal body physiological functions.

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