ANTICANCER ACTIVITY OF CYNANCHUM ACUTUM LEAVES EXTRACT ON WISTAR RATS TREATED WITH DIETHYL NITROSAMINE

 ANTICANCER ACTIVITY OF CYNANCHUM ACUTUM LEAVES EXTRACT ON WISTAR RATS TREATED WITH DIETHYL NITROSAMINE

Inyroduction

Cynanchum acutum belongs to the family Asclepiadaceae (milk-weed family) which comprises

about 2900 species in 315 genera (Boulos, 2000).

The Cynanchum genus comprises about 200 species (Boulos, 2000) reported for their use in folk medicine as antifebrile, antitumour, antitussive, diuretic, expectorant, anticonvulsant, anodyne, and tonic agent, and is effective against chronic hepatitis (Tawfiq, 1991). C. acutum, native to Southern Europe, is the only Cynanchum species mentioned in the Egyptian flora (Täckholm, 1974). Screening

of the biological activities of the total alcoholic extract of the underground organs of C. acutum revealed that it could inhibit the force and frequency of the intestine, heart and uterine contraction; it

also exhibited a marked anti-inflammatory effect

(Tawfiq, 1991). The alcoholic extract of leaves could be used as anti-inflammatory, analgesic,

antipyretic, molluscicidal, insecticidal, in the treatment of cardiac arrhythmia, intestinal colic, as hypotensive and for improving respiration in

asthma (El-Lithi, 1993; Abou Zeid et al., 2001;

Awaad, 2000).

The flavonoids of C. acutum were

0939Ð5075/2008/0900Ð0658 $ 06.00 ” 2008 Verlag der Zeitschrift für Naturforschung, Tübingen · Verlag der Zeitschrift für Naturforschung · D

tentatively identified by HPLC and chemical analysis (Abou Zeid et al., 2001; Heneidak et al., 2006),

whereas other phytochemical studies reported

the identification of some sterols (Halim et al.,

1990). In the present study, we report the isolation and identification of seven flavonoids from the butanol fraction (BF) of the methanolic extract of

C. acutum. Flavonoids are known to exhibit strong antidiabetic (Singab et al., 2005; Wang and Ng, 1999; Shukla et al., 2004; Chylack and Cheng,

1978; Hnatyszyn et al., 2002) and antioxidant activities (Rice-Evans et al., 1996; Heim et al., 2002;

Cao et al., 1997) which prompted us to test the BF and the major isolated flavonoids 1Ð3 for these effects.

METHODS

The alkylating agent DEN (diethyl nitrosamine) is

a well studied liver carcinogen, which when

administered continuously to rats, produces a well characterized dose–response relationship between

concentration in the drinking water and liver tumor

incidence (Druckcy et al., 1963; Lowry et al., 1951;

Peto & Park, 1985). Boucheron (1987) reported that a pro-mutagenic product accumulates in the hepatocyte of rats exposed to DEN in a dose-dependent manner.Single dose of diethyl nitrosamine (DEN) @ 200

mg/kg body weight was given to each rat of group II and

group III. The rats of group III were given herbal

formulation, daily p.o., from day 1 for 8 weeks; however,

the rats of group II were not given any kind of

treatment.

Weekly body weight was taken throughout the

experimental period to observe the effects on growth

rate. After 9 weeks of treatment all the rats were

sacrificed. The blood was collected after heart

puncture and 1 ml was taken in tubes containing

heparin (anticoagulant) and rest of the blood was kept in slant for clotting. The haematological examinations

performed were according to standard methods.

Erythrocytes and total leucocytes were counted using the improved Neubauer haemocytometer. Clotted blood samples were centrifuged at 5000 rpm, for 10mins, for serum collection. The serum samples were then aliquoted in vials and stored at –20°C. Serum samples were subjected to biochemical studies, namely aspartate aminotransferase/alanine

aminotransferase (AST), (ALT), alkaline phosphatase,acid phosphatase, bilirubin, total proteins, albumin,

cholesterol, High Density Lipid-cholesterol,

triglyceride, creatinine and uric acid etc. by using

Autopack kits as per manufacturer’s instructions.

Tissue homogenate of liver and kidneys were

prepared and subjected to assay of superoxide

dismutase (SOD) by following the method of Mishra & Fridouvich (1972), catalase activity (CAT) by Beers& Seezers (1952), Protein by Lowry et al., (1951), lipid acid phosphatase, bilirubin, total proteins, albumin,cholesterol, High Density Lipid-cholesterol,triglyceride, creatinine and uric acid etc. by usingAutopack kits as per manufacturer’s instructions.

Blood samples were

subjected for the estimation of tissue marker enzymes

like Serum glutamic oxaloacetic transaminase

(SGOT), Serum glutamic pyruvate transaminase

(SGPT), alkaline phosphtase (ALP), urea, and

creatinine by using Bayer’s Auto pack kits. The results were analysed statistically. Data was presented as mean ± standard error (SE). The difference between mean groups was assessed using F-test (one way analysis of variance ANOVA).

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