Effect of Aqueous Extract of Selected Plants on Liver, Bone and Kidney Markers of 7,12-Dimethylbenz(A)Anthracene (DMBA) Treated Albino-Rats

Introduction

7,12-dimethylbenz[a]anthracene(DMBA) is a polycyclic-aromatic hydrocarbon that acts as an immunosuppressant and it used to induce various cancers such as breast, lungs, oral and skin [1]. Cancer is one of the foremost causes of morbidity and mortality worldwide [2]. Cancer occurs when there is an uncontrolled cell division and proliferation.

Breast cancer (one of the cancers induced by DMBA) is the second most common cancer among women globally [1]. Breast cancer when developed, usually spread to the bones, lungs, and liver. Approximately 40–50% of women with metastatic breast cancer will have liver metastasis at some point during the course of their disease [3].

Chemotherapeutic treatment is used routinely for cancer treatment but some toxicity might be associated with chemotherapeutic drug usage. Plant-derived therapeutic products have been propounded for cancer treatment such as vinca alkaloids, taxanes, epippodphyllotoxins and camptothecin derivatives. Plants been a reservoir of natural bioactive compounds provide chemoprotective potentials against cancer [2]. The plants examined were Sorghum vulgare, Brillantaisia owariensis and Eremomastax polysperma. Sorghum vulgare is of the Poacea (grass) family. Sorghum vulgare leaf-sheath have been shown to contain phytate, tannins, cyanide, flavonoids [4], saponins, phenols and ribalinidine (an alkaloid) [5]. Whereas Eremomatax polysperma and Brillantaisia owariensis belongs to the Acanthaceae family. These species contain flavonoids, alkaloids, phenols, tannins, saponins and also have antioxidant properties [5, 6, 7]. Hence this study was aimed at determining the effect of aqueous extract of selected plants on the liver, kidney and bone markers of DMBA –treated rats.

Discussion

Cancer is a class of diseases in which a group of cells display uncontrolled growth, invasion and sometimes metastasis to other locations in the body via lymph or blood [13]. Breast cancer originates from the breast tissue, most commonly from inner lining of milk ducts (DCIS) or lobules (LCIS) that supply the ducts with milk which are prone regions for tumor microenvironment [1]. Breast cancer can metastasize to different organs of the body, especially tissues, such as, the bone, lung, liver and brain [14]. Thus determining liver and bone biomarkers is of significance. Table 1-4 demonstrates the consequences of Liver function test of DMBA treated albino rat, given aqueous concentrate of the different plants.

The AST activity of all groups were essentially diminished (p≤0.05) when juxtaposed with DMBA un- treated animals (63.33±8.56U/L) aside from those animals incited with tumour using DMBA and treated with 500mg/kg Eremomastax polysperma (73.66±2.33U/L). Whereas the ALT activity of all treated groups were significantly lower (p≤0.05) than DMBA untreated animals (30.66±0.66 U/L). ALT is a biomarker of hepatocellular injury and a rise in plasma level is indicative of an injury to the cytoplasmic membrane in the cell. AST is utilized to monitor the cause of various liver disorders [15]. This suggests the concentrates had no hepatoxicity. The ALP activity of all treated animals were significantly reduced (p≤0.05) when compared to DMBA untreated group (52.00±2.30 U/L) except for those animals induced with DMBA and treated with 500mg/kg Eremomastax polysperma (50.00±0.00 U/L). Elevated levels of Alkaline phosphatase may be due to intrahepatic obstruction or extrahepatic obstruction, or if it is present in serum in large amounts, it is diagnostic of bone or liver disease or a tumour in these organs [15]. This study indicates the extracts except 500mg/kg Eremomastax polysperma do not have effect on the enzyme activity, thus may not cause hepatic injury and possibly no metastasis to the bones. The ALT and AST. activities reduction is in accordance but in contrast in ALP activity with those of Wang and Zhang [16], which showed a significantly reduced ALT, AST and ALP activities when treated with Honokiol of DMBA-induced breast tumour animals. ArroyoAcevedo, et al.[17], additionally completed a comparable work utilizing P. auduncum case and demonstrated that it could lessen ALT activity and demonstrated no critical disparity in ALP activity.

The total bilirubin concentration of those animals induced with tumour using DMBA and treated with a standard drug (Tamoxifien) (13.60±0.60 μmol/L), 500mg/kg Sorghum vulgare leaf sheath (13.56±1.23 μmol/L), 1000mg/kg Sorghum vulgare leaf sheath (13.86±1.06 μmol/L), 1000mg/kg E. polysperma (10.43±0.82 μmol/L) and 1000mg/kg Brillantaisia owariensis (11.53±0.16 μmol/L) was significantly reduced (p≤0.05) when compared with DMBA animals (15.43±1.61 µmol/L).

But for conjugated bilirubin concentration, there was

no critical distinction (p≤0.05) in all groups aside from

those animals treated with 500mg/kg of Eremomastax

$$ p o l y s p e r m a \quad (1 3. 4 3 \pm 0. 6 9 \mu \mathrm {m o l} / \mathrm {L}) \mathrm {w h i c h} \mathrm {h a d} a $$

fundamentally high value (p≤0.05) when contrasted with

the different groups. Increased levels of total bilirubin

demonstrate jaundice [15]. This manner validates that the

concentrate possesses no deleterious impact on total

bilirubin and conjugated bilirubin concentration. The

total bilirubin concentration of this present study was

higher than Baccharis dracunculifolia (3.42µmol/l) [18]

but lower than Punica granatum (18.29-22.25µmol/l)

[19] treated mammary tumorigenic animals.

For the albumin and total protein concentrations,

there was no striking contrast (p≤0.05) within the groups

for the total protein concentrations. While there was huge

increment (p≤0.05) in albumin concentration of animals

actuated with DMBA and treated with 500mg/kg guinea

corn leaf sheath (37.00±1.15g/l), 1000mg/kg of Sorghum

vulgare leaf sheath (37.33±0.66g/l) and 500mg/kg

$$ E r e m o m a s t a x \quad p o l y s p e r m a \quad (3 8. 0 0 \pm 2. 3 0 g / l) \quad w h e n $$

contrasted with the control (32.33±0.88g/l) and DMBA-

untreated animals (31.66±0.88g/l). Other groups had

higher values than DMBA and the control groups,

although not significant (p≤0.05). Serum proteins, such as

albumin denotes malnutrition and identification of

response to inflammation. In breast cancer patients,

hypoalbuminemia does occur with values < 3.0g/dl [20].

This shows the plants concentrate may not pose hepatic

damage since the concentration of albumin and total

protein were within range and may be viable in hoisting

hypoalbuminemia that may happen in breast tumour

patients. The total protein and albumin concentration of

this study was similar to those of Kakehashi, et al. [18],

which demonstrated that total protein and albumin was

68-69g/l and 31-34g/l respectively, in tumorigenic wistar

rats. Although it was higher than those animals treated

with Punica granatum (albumin as 27.2-31.8g/l and total

protein as 42.4-54.6g/l)[19].

For GGT activity (Table 4), there was critical decline (p≤0.05) for those animals treated with 500mg/kg Brillantaisia owariensis (3.6U/L) when analogised with the control (4.43U/L) and the DMBA (5.53U/L) groups. There was no critical distinction (p≤0.05) among other groups, in spite of the fact that there was diminished values in the animals treated with 500mg/kg Eremomastax polysperma (4.83U/L) when compared to the DMBA (5.53U/L) animals. Elevated serum intensities of GGT are markers of oxidative stress (particularly in carcinogenesis) [21]. The diminished valuations of the treated group, shows the concentrates could decrease oxidative stress that would have been activated by DMBA harm. The GGT activity decrease is as per those of Wang and Zhang [16], which demonstrated a lessened GGT activity when treated with Honokiol of DMBA-instigated breast tumour animals.

The kidney work test of DMBA administered albino animals, given aqueous concentrate of the different plants is appeared on Table 5. There was no huge distinction (p≤0.05) in the uric acid concentration of all the groups aside from the animals treated with a standard medication (Tamoxifen), which had essentially higher values (p≤0.05) when juxtaposed with the control (Table

5). Uric acid is connected seriously with renal disappointment and cardiovascular illness, hypertension and metabolic disorder [22].

The creatinine and urea concentration was higher fundamentally (p≤0.05) in those animals treated with the standard drug and 1000mg/kg Eremomastax polysperma when compared to the DMBA (creatinine) and control (urea). The urea concentration of animals treated with 1000mg/kg Sorghum vulgare leaf-sheath and 500mg/kg Eremomastax polysperma was significantly reduced (p≤0.05) when compared to the DMBA untreated group and those groups treated with a standard drug (Tamoxifen). The reduction of urea concentration by the plants extract, supports the claims of Arroyo-Acevedo, et al. [17]; that P. auduncum capsule was able to reduce urea concentration when contrasted to the untreated group, in DMBA-induced mammary tumour. The creatinine concentration of all the groups in this study was higher than those of Soliman and Elfeky [23], which demonstrated the creatinine concentration of Ginger and cinnamon as 55.70µmol/l and 59.24 µmol/l respectively of breast cancer rats. Also, the uric acid concentration (84.46-94.57 µmol/l) of this present study was higher than the DMBA-incited animals treated with Holothuria atra [24]. Furthermore, it is similarity to the works of Ibrahim, et al. [25] for uric acid and urea concentration but dissimilar for creatinine concentration of animals incited with tumour using DMBA and treated with Brassica oleracea was lower than the DMBA untreated group.

Table 6 demonstrates the Electrolytes concentration of DMBA administered albino animals, given aqueous concentrate of the different plants. There was striking diminution (p≤0.05) in the Na+ concentration for the animals treated with 1000mg/kg Sorghum vulgare leaf sheath,1000mg/kg Eremomastax polysperma, and 1000mg/kg Brillantaisia owariensis. Low sodium concentration, below 120mEq/l is usually found in breast cancer patients [26]. This indicates no breast cancer since all the Na+ values were above 120mEq/l. For K+ concentration, the DMBA group had lower concentration however not huge (p≤0.05) when juxtaposed with the control, animals treated with standard medication, 500mg/kg S. vulgare group and 500mg/kg E. polysperma group. Hypokalemia do happen because of danger in breast malignancy patients while hyperkalemia is a marker of renal failure difficulty [27, 26]. The HCO3- concentration for all groups was within range (24- 30mmol/L) except for the DMBA-untreated group and the animals treated with standard medication which possessed lower than normal values showing acidosis, in

spite of the fact that there was no huge contrast (p≤0.05).

There was no striking variance (p≤0.05) in the chloride

concentration within the groups. In contrast to the works

of Kakehashi, et al. [18], who stated that the electrolyte

concentration of tumorigenic animals treated with

ethanolic extract of Bacchaaris dracunculifolia for Cl-

$$
(9 9. 1 \pm 3. 3 - 1 0

$$ (9 9. 1 \pm 3. 3 - 1 0 1. 1 \pm 3. 0 \mathrm {m E q} / \mathrm {L}), \text {a n d} \mathrm {N a} ^ {+} (1 4 2. 6 \pm 1. 9 - $$

142.1±1.9mEq/l) was lower than the results gotten from

this study, whereas the K+ (4.0±0.3-4.8±1.5mEq/L) was

higher than the results of this present study except for the

animals treated with 500mg/kg of E. polysperma which

had higher K+ value.

As earlier stated, one of breast cancer metastatic site is

the bones. Bone markers include alkaline phosphatase,

calcium and acid phosphatase. Table 7, demonstrates the

ACP (U/L) activity of DMBA administered albino rats,

given aqueous concentrate of the different plants. There

was noteworthy decrease (p≤0.05) in animals treated

with the standard drug when juxtaposed with the DMBA

un-treated animal and the control; other groups

$$ \text {demonstrated no huge difference (p \leq 0. 0 5). Acid} $$

phosphatase (AP) is a marker of prostate growth [28] and

of metastatic bone malady in patients with cancer of the

breast [29]. Normal serum range of acid phosphatase is

2.5 to 3.7ng/ml. From the study, there was no bone

metastasis since the AP activity of all the groups was

within the normal range. This study differs with those of

Gopikrishnan, et al. [30] which demonstrated that

Naringin was able to reduce Acid phospahtase activity of

DMBA induced mammary carcinogenic rats when

contrasted with the DMBA untreated group [31, 32, 33, 34, 35].

The Ca2+(mmol/l) concentration of DMBA treated albino rats, given aqueous extract of the various plants is shown on Table 7 also. There was significant reduction (p≤0.05) in Ca2+ concentration (which was below normal 2.0-2.6mmol/L) of animals treated with standard medication, 500mg/kg Sorghum vulgare leaf-sheath, 1000mg/kg Sorghum vulgare leaf sheath and 1000mg/kg Eremomastax polysperma when juxtaposed with the DMBA untended animal and the control. Indicating there might be slight kidney damage if not properly checked but there may not be bone metastases since there was no increase beyond the normal range. In contrast, Kakehashi, et al. [18] stated that the electrolyte concentration of tumorigenic animals treated with ethanolic extract of Bacchaaris dracunculifolia for Ca2+ was10.7±0.4- 10.6±0.4mEq/L, which is higher than all the treated groups in this study [35, 36, 37, 38, 39, 40, 41, 42].

Conclusion

In conclusion, the aqueous extract of the plants’ poses no hepatotoxic injury and was able to increase the albumin and total protein concentration.