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PROJECT TOPIC ON EFFECTS OF KOLAVIRON ON LYMPHOCYTES PROLIFERATION, EXPRESSION OF TOLL LIKE RECEPTOR-2 AND VASCULAR ENDOTHELIAL GROWTH FACTOR-C GENES IN Wuchereria bancrofti INFECTED BLOOD

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EFFECTS OF KOLAVIRON ON LYMPHOCYTES PROLIFERATION, EXPRESSION OF TOLL LIKE RECEPTOR-2 AND VASCULAR ENDOTHELIAL GROWTH FACTOR-C GENES IN Wuchereria bancrofti INFECTED BLOOD

ABSTRACT

The anti-proliferative effect and down regulation of vascular endothelial growth factor C and toll like receptor-2 by kolaviron rich portion of Garcinia kola on Wuchereria bancrofti infected blood were investigated. Blood samples were collected from consenting volunteers in Talata mafara, Zamfara State, Nigeria, between the hours of 10pm to 12am, stained with Giemsa and viewed microscopically for presence of microfilariae. Wuchereria bancrofti positive blood samples were cultured in vitro for 72 hours using untreated group as negative control, doxycycline (2µg/ml) treated samples as the positive control while the kolaviron (5µg/ml) treated group was the tested group. Mitotic index, expression of vascular endothelial growth factor-C (VEGF-C), toll like receptor- 2 (TLR-2) were determined using standard procedures, in peripheral blood lymphocytes. Mitotic index was significantly (P<0.05) reduced in the kolaviron treated group compared to negative control. Kolaviron also significantly (P<0.05) downregulated the expression of VEGF-C and TLR-2 when compared with the untreated group. In both cases, the effects of kolaviron was not significantly different (P<0.05) compared to that of doxycycline treated group. Furthermore, strong positive correlations (r) between mitotic index, VEGF-C and TLR-2 expressions were observed. The study concludes that kolaviron rich portion of Garcinia kola exhibited anti-proliferative effect and down regulation of VEGF-C and TLR-2 in Wuchereria bancrofti infected blood. Thus, the results from this study might have unravelled the potential of kolaviron in the management of complications associated with lymphatic filariasis.

CHAPTER ONE

INTRODUCTION

1.1 Background of the Study

Lymphatic filariasis, caused by parasitic Wuchereria bancrofti, is a mosquito borne disease characterized by a broad spectrum of clinical manifestation such as temporal/permanent disability and disfiguring leading to severe damage and painful swellings (lymphedema) of the legs and genitals in the late stage of the disease (Hoerauf et al., 2011; WHO, 2012; Gomase et al., 2013) and eventually stigmatization (WHO, 2013). Although the events leading to the development of chronic pathology in lymphatic filariasis are not fully understood, live filarial parasite and/or their productslive filarial parasite and/or their products have a direct effect on lymphatic endothelial cells and in the cells of the innate and adaptive immune system (Nutman, 2013). Vascular endothelial growth factor (VEGF) family which is key regulators of endothelial cell functions has been implicated in lymphangiogenesis and angiogenesis in lymphatic pathology (Pfar et al., 2009). Their levels are significantly elevated in individuals with filarial infection both in chronic and microfilaremic states (Bennuru et al., 2010). The key mediators when it comes to complications associated with lymphatic filariasis are toll like receptors (TLR). They are pattern recognition factors of the innate immune system responsible for the microbial detection and initiation of the host immune response (Kawai and Akira, 2010). Wolbachia, a Gram negative endosymbiont in filarial parasites are key inducers of pro inflammatory cytokines which interact with the immune system through TLR2 thus, contributing to the pathology of lymphatic filariasis (Hise et al, 2007).

The World Health Organization (WHO) initiated the Global Programme to Eliminate Lymphatic Filariasis with the goal to eradicate lymphatic filariasis in endemic countries. This initiative utilizes Mass Drug administration, where Diethylcarbamazine, Albendazole or Ivermectin is administered mass treatment of lymphatic filariasis annually in endemic areas (WHO, 2013). These drugs have been proven not to have good macrofilaricidal efficacy and reported resistance developed by the parasites against the drugs (resistance associated mutation at Tyrosine codon 200 of β tubulin of parasite in albendazole treatment) and they do not alleviate the pathology of the disease ( Schwab et al., 2005; Hoerauf et al., 2011).

Recently, doxycycline a broad spectrum anti-biotic from the tetracycline family spectrum anti-biotic from the tetracycline family ,has been used on a single daily dose of 200mg for six weeks in the treatment of Bancroftian filariasis. The treatment resulted in the reduction of plasma vascular endothelial growth factors ( Hoerauf, 2008; Bennuru et al., 2010), hence the need to use it as our positive control in this study.

Kolaviron is found in defatted ethanol extract of Garcinia kola. The extract is one of the numerous plant products that have been found to have a wide range of medicinal value which include anti-diabetic, anti-inflammatory and anti-proliferative effects amongst other ( Farombi and Owoeye, 2011; Ayepola et al., 2014).

In Nigeria, the use of medicinal plants cannot be over emphasized. Garcinia kola is cheaply sourced, readily available and often used for medicinal purposes. Kolaviron is yet to be explored in the management of lymphatic filariasis. Hence, the present study, for the first time, aims to report the role of kolaviron in managing the pathology associated with lymphatic filariasis by focusing on the possible anti-proliferative and the down regulation of toll like receptor 2 and vascular endothelial growth factor C in W. bancrofti infected blood lymphocytes. The findings from this work will go a long way in understanding the potential of kolaviron in the management of complications associated with lymphatic filariasis.

EFFECTS OF KOLAVIRON ON LYMPHOCYTES PROLIFERATION, EXPRESSION OF TOLL LIKE RECEPTOR-2 AND VASCULAR ENDOTHELIAL GROWTH FACTOR-C GENES IN Wuchereria bancrofti INFECTED BLOOD

1.2  Statement of Research Problem

Nigeria is the third most endemic country in the world with 22.1% prevalence of W. bancrofti infection (Nwoke et al., 2010).

Global Programme to Eliminate Lymphatic Filariasis is a mass drug administration intervention, involving the use of Ivermectin, Albendazole and Diethyl carbamazine. This only breaks transmission cycle (WHO, 2013) but do not alleviate the pathology of the disease.

These drugs are reported to have low effects and resistance to these drugs are also emerging (resistance associated mutation at Tyrosine codon 200 of β tubulin of parasite in albendazole treatment) ( Schwab et al., 2005; Hoerauf et al., 2011)

1.3 Justification for the Study

Lymphatic filariais is of great public health importance as it is one of the major caus of disability (WHO, 2013). The drugs used currently have little effect in ameliorating the pathogenesis of the disease hence the need to look for other means of management. Garcinia kola is cheap and locally sourced and understanding its effect on the immunopathology of lymphatic filariasis could be a stepping stone in the development of therapeutic drug in the management of Lymphatic filariasis or possibly in the production of target based DNA vaccine. There is no report yet on the effect of kolaviron on Lymphatic filariasis. The effect of Kolaviron on lymphatic filariasis is yet to be studied; since reports have shown its anti-inflammatory and anti-proliferative effect, kolaviron may be potent in the management of lymphatic filaraisis.

1.3 General Aim

To investigate the effects of Kolaviron on the immunpathology of Lymphatic Filariasis.

Rcommended: EFFECTS OF DIFFERENT TYPES OF ORGANIC FERTILIZERS ON GROWTH PERFORMANCE, NUTRIENTS AND TOXICOLOGICAL COMPOSITION OF AMARANTHUS CAUDATUS AND AMARANTHUS CRUENTUS

1.4 Specific Objectives

  1. To determine the anti–proliferative effect of kolaviron on the cultured peripheral blood lymphocytes of Wuchereria bancrofti infected blood using Mitotic index assay.
  2. To determine the effects of Kolaviron on the expression of toll like receptor-2 genes in the cultured peripheral blood lymphocytes sample using Real time-PCR.
  • To determine the effects of Kolaviron on the expression of vascular epithelial growth factor- C (VEGF C) gene in the cultured peripheral blood lymphocytes sample using Real time-PCR.

Seee also: ANTIDIARRHOEAL AND ANTIBACTERIAL ACTIVITIES OF METHANOLIC EXTRACTS AND FRACTIONS OF THE PULP AND SEED OF ZIZIPHUS MAURITIANA (CHINEE APPLE)

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