Box Nairobi , Kenya Corresponding author. Plants were collected from Baringo district, dried, extracted, weighed and tested for antileishmanial activity. Serial dilutions of the crude extracts were assayed for their activity against Leishmania major in cell free cultures and in infected macrophages in vitro. Inhibitory concentrations and levels of cytotoxicity were determined. Warburgia ugandensis, Psiadia punctulata and Chasmanthera dependens had an IC50 of 1.
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Box Nairobi , Kenya Corresponding author. Plants were collected from Baringo district, dried, extracted, weighed and tested for antileishmanial activity. Serial dilutions of the crude extracts were assayed for their activity against Leishmania major in cell free cultures and in infected macrophages in vitro. Inhibitory concentrations and levels of cytotoxicity were determined. Warburgia ugandensis, Psiadia punctulata and Chasmanthera dependens had an IC50 of 1. The supernatants from control and Leishmania infected macrophages were analyzed for their nitrite contents by Griess reaction and nitrite absorbance measured at nm.
Warburgia ugandensis stem bark water extract , Chasmanthera dependens stem bark water extract and Psiadia punctulata stem bark methanol extract produced All experiments were performed in triplicate. ANOVA was used to determine the differences between the various treatment groups. The analysis program Probit was used to determine IC50s. Keywords: Warburgia ugandensis, Psiadia punctulata, Chasmanthera dependens, anti-leishmanial activity, cytotoxicity and immunomodulative effects Introduction Leishmaniases are a group of primarily zoonotic diseases, caused by the trypanosomatids of the genus Leishmania that are transmitted by phlebotomine or Lutzomyia sand fly species Croft and Vanessa, Leishmaniases have an estimated million people at risk; incidence of two million new cases each year and a prevalence rate of 12 million people worldwide Ashford et al.
The global burden of leishmaniases caused a morbidity burden of 2. Visceral leishmaniasis is the most severe form of leishmaniasis, involving the internal organs and is fatal if left untreated Ashford and Bates, Unlike the other forms of leishmaniasis in which the parasite is histologically localized, VL is caused by parasite growth within reticuloendothelial cells throughout the body Boelaert et al. It is characterized by irregular fever, weight loss, hepatosplenomegaly and anaemia Manuel, Cutaneous leishmaniasis or oriental sores, produces skin lesions on the face, arms and legs, and is often self-healing Ashford and Bates, Mucocutaneous leishmaniasis a mutilating disease or espundia, as it is known in South America, causes disfiguring lesions to the face and destroys the mucous membranes of the nose, mouth and throat Walton, Reconstructive surgery of deformities is an important part of therapy Conjivaram et al.
Traditional uses of medicinal plants consist of oral administration of crude plant extracts for visceral leishmaniasis and as topical preparations of the corresponding extracts for the treatment of cutaneous leishmaniasis Croft and Vanessa, Natural products reported to have antileishmanial activity include quinones, alkaloids quinolines and isoquinoline analogues, indole analogues and steroidal alkaloids , terpenes iridoids, monoterpenes, sesquiterpenes, diterpenes, triterpenes and saponins , phenol derivatives chalcones and flavonoids and other metabolites such as acetogenins Manuel and Luis, In , a chloroform fraction CLF and a purified indole alkaloid obtained from crude stem extract of Peschiera australis was shown to have leishmanicidal effects against Leishmania amazonensis, a causative agent of cutaneous leishmaniasis in the New World.
In a bioassay-guided chemical fractionation, the leishmanicidal activity in CLF completely and irreversibly inhibited promastigote growth. This fraction was also active against amastigotes in infected murine macrophages. Chemical analysis of CLF identified an iboga-type indole alkaloid coronaridine as one of its major compounds. Promastigotes and amastigotes treated with CLF or coronaridine showed pronounced alterations in their mitochondria as assessed by transmission electron microscopy Delorenzi et al.
The recommended drugs used for the treatment of visceral leishmaniasis VL and cutaneous leishmaniasis CL , the pentavalent antimonials, were first introduced 60 years ago. Over the past decade alternative drugs or new formulations of other standard drugs have become available and registered for use in some countries, whilst other drugs are on clinical trial for both forms of the disease.
Although the ambition to develop a single drug or drug formulation effective against all forms of leishmaniases is unlikely to be fulfilled, the advances have been significant as the concept of choice for treatment is now real. Therefore, we set out to investigate whether extracts from W. Authentication was achieved by comparison with Herbarium specimens by taxonomists and a voucher specimen from each plant was deposited at KEMRI.
Plants extraction Stem bark and leaves at the secondary stage of growth were washed to remove physical impurities, air- dried, shred, and ground with an industrial blender and weighed. Water extraction was done using distilled water. Fifty grammes of ground powder was dissolved and soaked in ml of water for 3 hrs, placed in warm water bath for 1 hr, filtered and freeze dried.
Methanol extraction was done using distilled methanol as organic solvent, 20gms of ground powder was soaked overnight and concentrated using rotor evaporator. Two to 5gms of extract were obtained in each type of solvent and plant part. Cell free, macrophage and nitric oxide assays were used to test these extracts for their different effects. Stationary phase promastigotes cultures were used in this study. Promastigote forms of L.
The mortality rates of promastigotes were determined using Trypan blue stain exclusion principle whereby the promastigotes permeable to the blue dye were the dead ones. The parasites were counted using a Neubauer chamber. The 24 well plates were then wrapped with Para film and incubated in a plastic chamber at C for 2—4 days, making observations in the course of the incubation. The control group was incubated in the absence of the extracts.
Parasite viability was assessed before and after incubation by staining with Trypan blue and examining under a microscope. After 3 days the macrophages were harvested by anaesthetizing the mouse one at time with chloroform in an aesthesia jar with a lid, till it was unconscious, swabbing the abdomen with spirit then snipping the skin and pulling it to expose the peritoneum completely.
Four millilitres of washing media Hanks buffered salt solution HBSS was introduced into the peritoneal cavity and the abdomen softly massaged. A lot of this media was withdrawn using 19 gauge needle into a plastic centrifuge tube placed on ice, centrifuged at rpm for 10 minutes at 40C and the supernatant was aspirated leaving about 0.
An overdose of pentobarbitone sodium 1ml was used to euthanize the mice and their carcasses incinerated. Concentrations of macrophages was adjusted to live cells per ml. The negative control group was incubated in complete RPMI media while the positive control group was incubated in Pentostam. Both the experimental and the controls were incubated for further 48 hrs, after which the experiment was terminated, dried, fixed stained and macrophages counted.
Percentages of infected macrophages were determined by counting cells in triplicate slides. The study was carried out according to the institutional, national and international rules in regard to animal experiments and biodiversity rights. Infection of macrophages and nitric oxide production Mouse peritoneal macrophages were cultured in 24 well plates for 72 hrs.
Adherent cultured macrophages were washed once and re-suspended in fresh culture medium. After 4hrs, the wells were washed again to remove unphagocytosed parasites.
The infected macrophages were then incubated with the test compounds for further 72 hrs, after which the supernatants were tested for NO production levels. Griess reaction test The supernatants from control and Leishmania infected macrophages were analyzed for their nitrite content by Griess reaction Green et al.
One hundred microlitres of supernatant was put in the 96 well plates. Plates were then read at nm in an enzyme-linked immunosorbent assay plate reader Ignacio et al. The nitrate content of samples was determined by comparison of the nitrate standards and nitrate standard curve Green et al. Results Antileishmanial activity The water and methanol extracts of the stem bark of W. Mean mortality rates in P. Stem bark extracts in W. Table 1 Leishmanicidal effects of W. Plant species.
Chasmanthera dependens - Plant
Apply any organic fertilizer Chasmanthera dependens special feature This climbing rose can be trained to grow on walls, trellises, arbors, fences, arches, pillars, posts or other structures. Chasmanthera dependens uses The plant is used for ornamental purpose Medicinal Use: used in medicine for arthritis, joint pain, rhuematotism, The bark is chewed as a remedy for venereal discharges or as a general tonic for physical or nervous weakness in inflammatory and exhausting diseases Frequently Asked Questions What is another name for a Chasmanthera dependens? Chasmanthera dependens is commonly known as Climbing plant,surrounding vegetation How tall do Chasmanthera dependens grow? On average Chasmanthera dependens grows 6. A Chasmanthera dependens plant bears flowers of color Yellow When Chasmanthera dependens blooms? With enough light, Chasmanthera dependens may produce Yellow flowers in May to frost How hard is Chasmanthera dependens to grow?