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Short Communication - CYTOTOXIC, ANTIOXIDANT, ANTIMICROBIAL ACTIVITIES OF METHONOL CRUDE EXTRACTS OF CARDIA OBALIQUA (LINN.).

Byline: H. U. Khan, R. A. Khan and M. Ahmed

ABSTRACT

Cardia obaliqua was used traditionally in Pakistan for the treatment of diabetes, microbial infection and jaundice. In the present study we arranged to evaluate the antimicrobial, antioxidant and cytotoxic activities of Cardia obaliqua methanol extract. Cardia obaliqua was obtained from Surrani, District Bannu for preparation of crude methanol extract. A standard protocol of agar well diffusion method was used for characterization of various bacterial strains. Agar dilution method was subjected to investigate the antifungal activities. Death rate of brine shrimps was checked to investigate cytotoxic efficacy of the crude extract. The results showed that the crude extract significantly controlled the microbial growth as compare to control, which was in agreement with the spacious use of tested plant samples in primary healthcare center. Similarly the extract showed significant potency in scavenging of free radicals.

From the present study it is inferred that Cardia obaliqua methanol extract was act as a vital role for the detection of new antimicrobial drugs and antioxidant agents; which confirm its conventional uses and its management as beneficial mediator.

Key words: Cardia obaliqua, Methanolic extract, Cytotoxic activity, Antioxidant activity, Antimicrobial activity.

INTRODUCTION

In the recent age human being throughout the globe is suffering from various infections which cause a large number of diseases. To overcome such miserable condition various strategies are being in process. Among them medicinal plants play crucial role in care the improvement of human health (Bruneton, 1955). In this regard ethano medical knowledge of traditional medicinal plants and their pharmacological screening are in practice in various regions of Pakistan (Khan et al., 2016). Medicinal plant contains a variety of potent bioactive ingredients which provide control and immunity to living things against various harmful agents (Khan et al., 2016). Medicinal plant contains a large number of antioxidants compounds which are used for the prevention of oxidative stress and protract the life of the functional substance.

For this purpose in the present, it is decided to add the antioxidants in suitable amount in the food as an additive, in cosmetics and in pharmaceutical products (Sahreen et al., 2015). According to different reports it was shown that various parts of medicinal plants viz; fruits, leaves, oil seed, roots and vegetables have free radical scavenging and microbial growth control efficacy (Rababah et al., 2004) which may be used as natural antioxidants and antibiotics (Sahreen et al., 2014). Cordia obaliqua plant are found worldwide but mostly in warmer regions of the world, producing edible fruits, known as a clammy cherries, glue berries, sebesten, or snotty gobbles and lasora. The fruits are also used as a vegetable and pickle. Cordia obaliqua is used as an ornamental plant and its wood is commercially harvested. According to binomial nomenclature Cordia obaliqua belong to Boraginaceae.

Cardia oblica is a local plant of District Bannu which is commonly used for the control of various human ailments without pharmacological confirmation. The present project was basically designed for the pharmacological evaluation via different assays viz; antioxidant, cytotoxic and antimicrobial activities of Cardia oblique crude methanol extract.

MATERIALS AND METHODS

Cardia obaliqua fresh leaves were obtained from Landidak area of District Bannu during fruiting sessions. After shad drying of fresh parts at room temperature (250C), it was ground mechanically to get fine powder. Among powder, 1 kg of Cardia obaliqua fine powder was socked in 70% methanol (1.5 L) and shacked randomly. After 96 hrs, the extract was filtered via Whatman filter paper No.1. Filtrate was concentrated using rotary evaporator at 38AdegC to get methanol crude extract (COME) and was kept at 4AdegC in the refrigerator for further in vitro investigation.

Cytotoxic brine shrimp lethality test was carried out according to standard protocol with some modification as described by Khan et al., (2015) for the determination of cytotoxic activity. DPPH (1, 1-diphenyle-2-picryle hydrazyl) scavenging activity was conducted using the procedure of Gymafi et al., (1999) to show antioxidant efficacy of crude methanol extract. To check the antifungal activity of the methanol crude extract of Cardia obaliqua, the standard protocol of Duraipandiyan and Lgnacimuthu (2009) was used.

Antibacterial activity of Cardia obaliqua methanol crude extract was carried out by the protocol of Bagamboula et al., (2003).

RESULTS

Prime screening of the extract through cytotoxic activity provides useful information about the antitumor and anticancer activity for the future use of the medicinal plants extract. Cytotoxic effect of the Cardia obaliqua methonolic crude extracts was measured and noted against brine shrimps growth as shown in Table 1. From the Table 1, it is clear that 50ug/ml treatment of crude extract showed 90% survival and 10% death, however at 100, 250, 500 and 1000 (ug/ml), there was 70, 50, 30 and 20 (%) survival and 30, 50, 70 and 80 (%) death occurred respectively which may be due to the presence of bioactive constituents present in the plant extract.

1,1-diphenyl 2-picryl - hydrazyl (DPPH) have the aptitude to obtain electrons from the antioxidants, that is why it is broadly utilized for the in vitro antioxidants scavenging assays of the medicinal plants. Table 2 shows the % scavenging activities of COME and ascorbic acid against the DPPH free radicals. By using the crude methanol extract with the increasing concentration such as 50<100<150<500<1000 (ug/ml), significant scavenging activities were observed viz; 54.6+-2.6<58.0+-3.1<65.5+-2.9<77.9+-3.8<89.1+-3.4.

Similarly ascorbic acid showed potent DPPH free radicle scavenging activities which was used as a standard.

Cardia obaliqua methanol crude extracts revealed low activity (29.78 %) against Aspergillus fumigatus followed by Aspergillus flavus (38.60%) while the highest activity was shown against Aspergillus niger (49.44%) and Fusarium salmoni (45.49%) as shown in Table 4. Similarly the terbinofine, a positive control has indicated high activity against these fungal strains. Terbinofine shows low activity (84.38%) against Fusarium salmoni followed by Aspergillus Flavus (85.73%) in comparison highest activity was found against Aspergillus niger (90.50%) and Aspergillus fumigatus (87.23%). DMSO (negative control/normal control) indicates zero percent (0%) inhibition activity against all the used four fungal strains.

COME showed low activity (13+-1.5mm) against Escherichia coli followed by Staphylococcus aureus (16+-2.4mm). The methanolic extract exposed highest antibacterial activity against Micrococcus luteus (21 mm) while comparatively high activity was observed against Pseudomonas aeruginosa (17+-2.3 mm) as shown in Table 5. Highest activity was observed against Staphylococcus aureus (21+-1.4) and in comparison high activity was observed against Escherichia coli (17+-2.7 mm), while highest activity was observed against Micrococcus luteus (23+-2.0 mm) and comparatively high activity was found against Pseudomonas aeruginosa (21+-1.8 mm) as shown in Table 5.

Table 1. Effect of Cardia obaliqua methonolic against brine shrimp assay.

Concentration of plant extract###Total No of brine###Survived brine shrimps###% Survival###% Death

###shrimps

50g/ml###10###9###90###10

100g/ml###10###7###70###30

250g/ml###10###5###50###50

500g/ml###10###3###30###70

1000g/ml###10###2###20###80

Table 2. % DPPH free radical scavenging activity

Concentration (in ug/ml)###%COME Scavenging###% Ascorbic acid Scavenging

50###54.6+-2.6###63.2+-2.5

100###58.0+-3.1###67.1+-2.8

150###65.5+-2.9###71.6+-3.9

500###77.9+-3.8###93.9+-4.6

1000###89.1+-3.4###97.1+-5.0

Table 3. % ABTS free radical scavenging activity

Concentration (in ug/ml)###%COME Scavenging###% Ascorbic acid Scavenging

50###35.17+-2.1###46.25+-2.1

100###42.11+-2.3###51.65+-4.1

150###55.06+-1.7###63.20+-3.7

500###69.77+-1.8###85.10+-2.5

1000###80.35+-2.4###95.96+-3.1

Table 4. Antifungal activity of Cardia obaliqua methanolic extracts.

Sample###Antifungal activity in term of percentage (%) inhibition

###Aspergillus niger###fusarium salmon###Aspergillus Flavus###Aspergillus fumigatus

COME (200 ug/ml)###49.44+-2.5###45.49+-6.1###38.60+-3.4###29.78+-2.0

Terbinofine (200 ug/ml)###90.50+-2.1###84.38+-5.1###85.73+-5.6###87.23+-5.1

Table 5. Antibacterial activity of Cardia obaliqua methanolic extracts (COME)

###Micrococcus luteus###Escherichia coli###Pseudomonas aeruginosa###Staphylococcus aureus

Concentration (mg/ml)###Zone (mm)###Zone (mm)###Zone (mm)###Zone (mm)

1 mg/ml cefix###16+-1.9###27+-2.4###18+-2.0###29+-2.4

1 mg/ml Ampicil###23+-2.0###17+-2.7###21+-1.8###20+-2.3

1 mg/ml COME###20+-1.2###19+-1.5###15+-1.6###14+-1.7

3 mg/ml COME###22+-1.3###18+-1.9###14+-2.1###13+-1.8

5 mg/ml COME###21+-1.4###13+-1.5###17+-2.3###16+-2.4

DISCUSSION

Reactive oxygen species (ROS) in lesser quantities are useful for the normal metabolism and signal transduction but in large amounts produces oxidative stress which outbreaks and harm numerous units like protein, lipids and DNA (Halliwell and Gutteridge, 1999). Medicinal floras showed crucial role in the cure of diverse mortal diseases viz; free radicals stress, inflammatory disorder, cardiac failure and metastasis in the biosphere from very long time. Diverse portions of natural products are recycled frequently for the handling of numerous sicknesses comparatively to the drugs prepared in various pharma industries. Pakistan is a rich state in therapeutic floras which are locally used in folk medicines for the treatments of various diseases like infections, cardiovascular diseases, diseases of digestive system and skin by local healer (Khan et al., 2016). The results got from the current study of Cardia obliqua (Linn) methanol extracts display important scavenging efficacy.

The antioxidant activity of the methanol extracts of the medicinal florae is due to the phenolic and polyphenolic compounds present in them, which distinctly decrease the free radicals (Shah et al., 2017). Our findings reveals some resemblances with the examination of Hagerman et al. (1998) and Falleh et al. (2008) who stated that free radicals are distinctly scavenge by medicinal plants. The consequences got from our trials are also stayed by the conclusions of Duenas et al. (2006) and Kilani et al. (2008). As fungi yield mycotoxin which meaningfully affect human health. Some fungus abolishes our foods and interrupts their dietetic value thus made them useful for our routine (Sun et al., 2002). In our current study the antifungal activity of Cardia obliqua (Linn), confirmed that fungal strains are considerably repressed by them.

Numerous reports of the medicinal floras for microbial actions designated that their actions are due to the occurrence of phenolic compounds and saponins (Aboaba and Efuwape 2001, Baydar et al., 2004, Rodringuez et al., 2007; Mohanta et al., 2007).

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Publication:Journal of Animal and Plant Sciences
Date:Oct 31, 2017
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