Project Materials

ELUCIDATION OF SOME IMMUNOLOGICAL AND BIOCHEMICAL NATURE OF THE LEAVES OF SENNA MIMOSOIDES

ABSTRACT

The phytochemical composition, immunomodulatory, leukocyte mobilisation, haematological, and antihepatotoxic properties of Senna mimosoides leaf aqueous extract were investigated in this work. The study also looked at the effect of the extract on lactase activity, the harmful effect of carbon tetrachloride (CCl4), and the extract’s ameliorative effect on liver tissue using histopathology. This study sought to validate the traditional use of S. mimosoides leaves in folkloric medicine to treat breast milk toxicity in neonates by explaining their immunological and biochemical properties. The phytochemical composition included 2.67 ± 0.0013 mg of flavonoids, 3.43 ± 0.0028 mg of alkaloids, 1.97 ± 0.0030 mg of saponin, 2.32 ± 0.0032 mg of terpenoids, 0.86 ± 0.0023 mg of steroid, 3.61 ± 0.0025 mg of phenol, 8.31 ± 0.0032 mg of reducing sugar, 4.75 ± 0.0034 mg of tannin, 1.61 ± 0.0031 mg of cyanide, 2.75 ± 0.0029 mg of glycoside, and 4.68 ± 0.0034 mg of tannin. The animal model experiment used one hundred and thirty (130) albino rats. The experimental design was divided into four (4) phases, each consisting of five (5) rats. Rats in group A (control) were given 0.2 ml of normal saline; rats in groups B, C, and D were given 50, 100, and 250 mg/kg of the aqueous extract of S. mimosoides leaves, respectively; rats in group E were given levamisol or silymarin (standard drugs); and rats in group F were given only carbon tetrachloride (CCl4). Administration of the extract at 50, 100, and 250 mg/kg resulted in a dose-dependent substantial (p < 0.05) increase in primary antibody titre (6, 8, 13) and secondary antibody titre (11, 26, 34). The delayed type hypersensitivity (DTH) reaction indicates that the extract caused a dose- and time-dependent increase in footpad oedema in rats. The extract (50, 100, and 250 mg/kg) and levamisol (25 mg/kg) significantly (p < 0.05) increased DTH reactions at 24 hr after challenge, with 1.412, 1.504, 1.816, and 1.827 mm difference in footpad thickness before and 24 hr after challenge, respectively. The control showed a non-significant (p > 0.05) increase of 0.614 mm. Footpad oedema increased by 1.908, 1.918, 2.304, and 2.326 mm for the extract and levamisol 48 hours after the challenge. The humoural antibody (HA) titre and DTH reaction are comparable to those of levamisol, a typical immunostimulatory medication, at 25 mg/kg. The total leukocyte count of the groups treated with different amounts of extract grew in a dose-dependent manner, while the group treated with indomethacin fell significantly (p < 0.05) compared to control. Group B had a percentage packed cell volume (PCV) of 38.8 ± 1.30 before and after treatment with cyclophosphamide (CP) and subsequently with (50 mg/kg), respectively. Groups C, D, and E followed the same pattern, but in the control group, the CP decline was not reversed. The control group had a percentage PCV of 35.8 ± 0.45 before CP and 19.4 ± 0.55 after extracting. The same pattern was seen in haemoglobin concentration, white blood cell count, red blood cell count, and associated indices. Rats in group F showed higher serum alanine aminotransferase (ALT) activity (81.20 ± 0.84 IU/L) following CCl4 injection compared to the normal control A (53.00 ± 1.00 IU/L). The extract (50, 100, 250 mg/kg) and silymarin (25 mg/kg) significantly reduced ALT activity (p < 0.05) to 65.00 ± 1.58, 59.20 ± 0.84, 55.20 ± 1.30, and 57.00 ± 1.00 IU/L, respectively. When compared to the control group, the levels of aspartate aminotransferase (AST), alkaline phosphatase (ALP), bilirubin, malondialdehyde, iron, and phosphate followed the same trend. Administration of CCl4 decreased reduced glutathione levels in group F (2.21 ± 0.239 mMol/g tissue). Treatment with varying amounts of the extract and levamisol increased the reduction (3.08 ± 0.093, 4.17 ± 0.241, 5.16 ± 0.193, and 4.97 ± 0.273 mMol/g tissue, respectively). The activities of glutathione s-transferase, glutathione peroxidase, catalase, and superoxide dismutase, as well as the amounts of sodium, magnesium, potassium, calcium, zinc, and selenim, followed the same trend. Histopathological tests revealed that the extract and levamisol improved centrilobular degeneration of the liver tissues caused by CCl4. Furthermore, the extract showed considerable (p < 0.05) lactase activity in a dose-dependent manner compared to the control. At 10, 20, 30, 40, and 50 µl, enzyme activity was 17.187, 18.8 22, 20.044, 22.022, and 23.898 IU. The results of this investigation indicate that the numerous medicinally relevant bioactive components found in this extract may be responsible for the immunostimulatory, antihepatotoxic action, rise in lactase activity, and haematological parameters. This supports the usage of this herb in folkloric medicine to treat ailments.

Chapter one

INTRODUCTION

Plants are known to have a wide range of secondary metabolites. These secondary metabolites, or bioactive substances, have specific physiological impacts on the human body.

According to Yadav and Agarwala (2011), plant-based compounds account for approximately 25% of all prescribed medicines today. Interestingly, several phytochemicals have been found and even isolated from a range of therapeutic plants. However, many of them have yet to be used in clinical settings.

Phytochemical analysis of plants is significant because of the expensive cost of synthetic medications, which necessitates the development of alternative plant-based drugs.

When these secondary plant metabolites extractable by various solvents are consumed by animals, they demonstrate a variety of biochemical and pharmacological activities (Nwogu et al. 2008).

The rationale for this study was the usage of Senna mimosoides in traditional medicine, specifically in Ukehe, Nsukka, to treat neonatal oedema and breastfeeding toxicity. Ekwueme et al. (2011a,b) investigated the anti-inflammatory properties of Senna mimosoides leaf extracts and their mechanisms of action.

To test the toxicity of breastmilk in Nsukka, it is frequently dropped on cocoyam leaves or ants immediately after delivery. Toxic breastmilk frequently burns the leaves of the cocoyam or kills any ants it comes in contact with.

The predominance of businesses exposes mothers to toxins that can collect in breast milk. In this work, the immunomodulatory activity and anti-hepatotoxic impact of S. mimosoides leaf extract were explored because these are the basic mechanisms employed by the body to prevent or cure diseases.

Furthermore, the effect of the leaf extract on the activity of lactase, the enzyme responsible for the degradation of lactose, the only carbohydrate found in breast milk, was investigated.

1.1 Overview of the Human Immune System.

Immunology is the study of the body’s defence mechanisms against infectious pathogens and other foreign substances in its environment (Wotherspoon, 2012).

The immune system contains thousands of components, and it appears that the immune system is significantly more intricate than is required to achieve what appears to be a straightforward task of eradicating a harmful organism or abnormal’self’ cells (Parkin and Cohen, 2001).