INVESTIGATING EFFECT OF FREEZE DRIED ROOT EXTRACT OF RHOICISSUS REVOILII (Bushveld grape) ON SMOOTH MUSCLE OF RABBIT UTERUS Proposal presentation By Bonface Okeyo Ochola BIO-1-0174-1/2014 Supervisors Peter Gitonga

DESCRIPTION OF RHOICISSUS REVOILII INTRODUCTION DESCRIPTION OF RHOICISSUS REVOILII Botanical name of the plant is Rhoicissus revoilii. It is a climber plant The leaflets are glossy green above, and reddish-brown Leaves 3-foliolate, ovate, with lateral leaflets strongly asymmetrical, densely covered in rusty or grey-green hairs.

PICTURE OF RHOICISSUS REVOILII PLANT

IN KENYA THE DISTRIBUTION OF RHOICISSUS REVOILII IS AS FOLLOWS

MEDICINAL USE OF RHOICISSUS REVOILII Rhiocissus revoilii is one common herb used in threatening abortion . Rhoicissus revoilii is used as an anaesthetics and painkiller It is used for to treat skin diseases such as sores In livestock R. revoilii is also used in treatment of lactation problem Rhoicissus revoilii, used in treatment of microbial infections .

Specific Objective RESEARCH OBJECTIVE General objective To investigate the effect of Rhoicissus revoilii freeze dried root extract on the contractility of isolated rabbit uterus preparation Specific Objective To determine the effect of R. revoilii freeze dried root on the contractility of the uterus. To investigate the dose-response effect of R. revoilii freeze dried root extracts on uterus contractility

STUDY QUESTION The proposed question is that, what effect does Rhoicissus revoilii Freeze Dried Roots have on the uterus smooth muscle contraction. Null hypothesis Rhoicissus revoilii Freeze dried root extract has no effect on uterus smooth muscle contractility. Alternative hypothesis Rhoicissus revoilii Freeze dried root extract has an effect on uterus smooth muscle contractility

LITERATURE REVIEW MISCARRIAGE Miscarriage, also known as spontaneous abortion and pregnancy loss, is the natural death of an embryo or fetus before it is able to survive outside independently The most common symptoms of a miscarriage are vaginal bleeding In those under the age of 35 the risk is about 10% while it is about 45% in those over the age of 40 About 5% of women have two miscarriages in a row

FRESH ISOLATED RABBIT UTERUS

The plant is collected along the edge of Ngong forest Nairobi RESEARCH METHODOLOGY The plant is collected along the edge of Ngong forest Nairobi The identity is confirmed at the university of Nairobi herbarium, Department of Botany. They roots will be washed and then dried then milled at the department of Chemistry University of Nairobi Resulting filtrate is deep frozen using a deep freezer located at the Department of Physiology University Nairobi overnight. The frozen filtrate is freeze dried at Kenya Medical Research Institute(KEMRI)

SUBSTANCE AMOUNT(5g/l) KCL 2.1 NACL 4.5 NAHCO3 2.5 D-GLUCOSE CACL2 0.4 COMPOSITION OF DE JALON SOLUTION SUBSTANCE AMOUNT(5g/l) KCL 2.1 NACL 4.5 NAHCO3 2.5 D-GLUCOSE CACL2 0.4

LABORATORY ANIMALS Female virgin adult New Zealand white rabbit (2.0-2.5kg) are to be used in the in vitro study. In preparation for uterus isolation three virgin female rabbits ranging from 22-24 weeks are injected with 0.2mg diethylstilbestrol (DES) (sigma Aldrich) intramuscularly 24 hours prior to the experiment ETHICAL CONSIDERATIONS The animals will be handled in accordance with Federation of Europeans Laboratories Animal Science Association FELASA guidelines

A POWER LAB DATA ACQUISITION SYSTEM

EXPERIMENTAL PROTOCOL A segment of uterus about2cm is cut and suspended in an 80ml organ bath filled with De jalon solution. The tissue is first allowed to equilibrate for 60 minutes while changing the physiological salt solution (PSS) every 20 minutes Effects of different concentration of RERR (0.5, 1.0, 2.0, 3.0 and 5.0mg/ml) on the spontaneous contraction of uterus is tested non-cumulatively. The rate and force of uterus contraction is recorded and analyzed using chart5 software AD instrument) To try and elucidate the possible mechanism of action, the effect of 1mg/ml is tested in presence of standard agonist; salbutamol

DATA ANALYSIS AND LIMITATION Analysis of variance will be used to test whether there will be statistical significance in contractility differences observed before and after administration of Root Extract of Rhoicissus Revoilii (RERR). A P-value of <0.05 will be taken as an indicator of a significant outcome limitaion Content of active ingredients in the roots of Rhoicissus revoilii can range significantly due to the growth and quality of preparation There is a small sample size

ITEM UNIT PRICE(KHS) COST PROJECT PREPARATION 1800 PRINTING 300×4 1200 BUDGET ITEM UNIT PRICE(KHS) COST PROJECT PREPARATION 1800 PRINTING 300×4 1200 PHOTOCOPYING 600×2 APPARATUS 3000 BINDING 300 RABBIT 1000×4 4000 CONTINGENCY 500 REAGENT 1000 TOTAL 13000

TIMELINES ACTIVITY/MONTH SEP –DEC2015 JAN 2016 FEB 2016 MARCH2016 APRIL 2016 PROPOSAL DEVELOPMENT PILOT STUDIES DATA COLLECTION AND ANALYSIS THESIS WRITTING PRESENTATION

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