INTRODUCTION:

The plant Hibiscus Rosa-sinensis belongs to the family Malvaceae. Many species of Hibiscus are planted mainly for showy flowers or landscape shrubs. It is widely cultivated in the tropics as an ornamental plants and has several form depending on the color of flower. It is also used for sometimes in paper making and was used as an anti asthmatic traditionally¹. The leaves and flower has an healing capacity². They are also found to promote hair growth. Many species of Hibiscus have been found to have a biologically active compounds which acts as an antioxidant, as a cardio-protective and are able to decelerate the proliferation of malignant cells. Many ingredients have been isolated from plants such cyanidin, quercetin, hentriacontane, hibiscetin, glycosides, carotene, taraxeryl acetate, β-sitosterol, campesterol, stigmasterol, ergosterol, citric, tartaric and oxalic acids, cyclopropenoids and anthocyanin pigments, calcium oxalate, thiamine, riboflavin, niacin and ascorbic acids³. It is extensively used as an antidote for research purposes in rodents against chemicals such as acids, alkali and pesticides. Hence, Hibiscus sp. are a great natural source for the development of new drugs and may provide a cost effective mean of treating cancers and other diseases in the developing world⁴.

It’s pharmacological studies has deciphered its antioxidant⁵, antidiabetic⁶, hepatoprotective⁷ cardioprotective⁸, antipyretic, anti-inflammatory⁹ and neuroprotective¹⁰ advantages. Hibiscus show strong antioxidant and antitumor promotion effects. Hibiscus extract is an apoptotic inducer and a specific activator of JNK/ p38 MAPK pathway. The molecular mechanism underlying this effect could be described as the induction of apoptosis via activation of the p38 MAP kinase that subsequently phosphorylates the target protein c-jun and trigger the signal to further activate the apoptotic protein cascades that contain Fas-mediated signalling. As an outcome, cytochrome c is released from the mitochondria, leading to the cleavage of caspase-3¹¹.

MATERIALS AND METHOD:

Test Animal:

A total of 30 young, healthy male rats of Charles foster strain were employed in the study on the basis of their initial health check-up and body weights, ranging between 125-150 grams. A maximum of five rats were kept in each cage. They were acclimatized to the laboratory conditions for seven days prior to the experiment in a well ventilated animal house under constant environmental and adequate nutritional conditions throughout the experiment. They were fed balance rodent pellet diet and water ad libitum.

The housing conditions were carried out according to GLP norms, Govt. of India. The animals were kept in sterilized polypropylene cages with 12 hours light (8:00 am-8:00 pm) and 12 hours dark cycle (8:00 pm-8:00 am). Temperature was maintained at 24 ± 20C with a relative humidity of 40 to 60% and 10-15 times fresh air changes per hour. The experiment on animals were approved and carried out according to the guidelines of Institutional Animal Ethics Committee (IAEC).

Figure 1: Male Albino Charles Foster Rat

Composition of Rat pellet diet is given below:

Crude protein 18 – 19%

Ether extract 05 – 06%

Crude fibre 5.0%

Ash Content 8.0 – 9.0%

Calcium 0.5 to 1.0%

Phosphorus 0.6 to 0.8%

Nitrogen free extract 50.0%

Metabolisable energy (Cal/Kg) 3600 (Soya Based)

Figure 2: Rat Pellet and Water ad libitum

Experimental Design:

The study was conducted following Schedule Y, Appendix III, India. Rats were divided into five groups with six rats per group, randomly. Group 1 was marked as control while group 2, 3, 4 and 5 were marked as treated and was administered only on the 1^st day of dosing. The animals were observed throughout the study for mortality and gross morphological, physiological, behavioural changes. The biological parameters were investigated terminally at the end of the study.

Dose Group		No. of Rats/ Group	Dose Range
Group I	Control (Distilled water)	6	0mg/kg
Group II	Treated (Hibiscus Rosa-Sinensis Extract + Distilled Water)	6	500 mg/kg
Group III	Treated ( Hibiscus Rosa-Sinensis Extract + Distilled Water)	6	1000 mg/kg
Group IV	Treated ( Hibiscus Rosa-Sinensis Extract + Distilled Water)	6	1500 mg/kg
Group V	Treated ( Hibiscus Rosa-Sinensis Extract + Distilled Water)	6	2000 mg/kg

Mode of Administration:

The freshly prepared Hibiscus Rosa-Sinensis Extracts with distilled water were administered orally by cannula for 14 days. Group I received distilled water while Group II, III, IV and V received Hibiscus Rosa-Sinensis Extracts + distilled water at a concentration of 500 mg/kg, 1000mg/kg, 1500 mg/kg, 2000mg/kg respectively in the same manner.

Figure 3: Oral Drug Administration to Rat

Gross Examination:

At the end of study, all the rats belonging to the different groups were sacrificed by exsanguination under light ether anaesthesia. Necropsy included the examination of the external surface of the body, all orifices, and the cranial, thoracic and abdominal cavities, and their contents. A thorough naked eye examination of size, shape, surface, contours etc. of all the important tissue and organs was done. Complete gross post-mortem examinations were performed on all terminated and dead animals.

Figure 4: Autopsy and Complete Gross Post-mortem Examination of Sacrificed Rat

Organ Weight:

Major organs and tissues (adrenal, brain, gonads, heart, kidney, liver, lung and spleen) were taken out from all the sacrificed rats. The absolute weights of each organ were measured and their relative organ weights (organ/body weightx100) were calculated.

Histopathological Examination:

Representative pieces from all the above-mentioned organs and tissues were preserved in 10% formalin solution to fix them. They were embedded with paraffin, sectioned, prepared slides and stained with haematoxylin and eosin (H and E method) and observed microscopically for histopathological examination.

RESULT:

Hibiscus rosa sinensis flowers are remarkably safe to use. No toxicity or side effects are expected in reasonable doses. In this work we use 60 healthy Charles foster rats for our experiment; we use 30 male rats and remaining 30 as female. In experiment rats divided into a group as control and treated group. Every group contain 6 rats’ male cage and female cage. Ist group keep as control group and remaining 5 group as treated group. The dose of extract was 500mg/kg, 1000mg/kg, 1500mg/kg and 2000mg/kg. the duration of the extract treatment were gradually increased from group IInd to group Vth . The rats were sacrificed accordingly after the last dose by the way our experiment we use only one time dosing and cheque the any type of toxicity in rats. The testes of the control groups were similarly collected at regular interval and historical examination done accordingly. Our study definitely revealed that the extract of the Hibiscus rosa sinensis show nontoxic effect in Charles foster rats.

There was no significant gain or loss in the body weight of the rats as observed throughout the experiment. There were no regular variations in the average 24-hour water and food intake of the rats of both control and treated group monitored. No mortality was seen in any of the groups either control or treated. The initial body weight of rats is slightly increase both control and treated animals but no significant difference were seen in the treated groups. In haematology parameters of Hibiscus rosa sinensis treated animals there is no reduction were seen in the values of T-RBC, no reduction were seen in the values of Hgb. The decreasing level of RBC and Hemoglobin showed anemic condition in rats

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Received on 10.02.2014 Modified on 22.02.2014

Res. J. Pharmacognosy & Phytochem. 6(1): Jan.-Mar. 2014; Page 37-40