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Possible Mechanism of Action of the Hypotensive Effect of Peperomia Pellucida and Interactions Between Human Cytochrome P450 Enzymes Chukwuemeka R

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Possible Mechanism of Action of the Hypotensive Effect of Peperomia Pellucida and Interactions Between Human Cytochrome P450 Enzymes Chukwuemeka R Arom & at al ic in P l ic a n d t Nwokocha et al., Med Aromat Plants 2012, 1:4 e s M Medicinal & Aromatic Plants DOI: 10.4172/2167-0412.1000105 ISSN: 2167-0412 ResearchResearch Article Article OpenOpen Access Access Possible Mechanism of Action of the Hypotensive Effect of Peperomia pellucida and Interactions between Human Cytochrome P450 Enzymes Chukwuemeka R. Nwokocha1*, Daniel U. Owu1, Kelece Kinlocke1, JeAnn Murray2, Rupika Delgoda2, Karen Thaxter1, Garsha McCalla1 and Lauriann Young1 1Department of Basic Medical Sciences, University of the West Indies, Mona Campus, Kingston 7, Jamaica 2Natural Products Institute, University of the West Indies, Mona Campus, Kingston 7, Jamaica Abstract Background: Peperomia pellucida is used as a medicinal plant and as an antihypertensive remedy. We investigated the possible mechanism of this action and its impact on cytochrome P450 (CYP) enzyme activity. Methods: Mean arterial pressure and heart rate were recorded via cannulation of the carotid artery on anaesthetized, normotensive Sprague-Dawley rats following intravenous administration of Peperomia pellucida aqueous (10-30 mg/ kg) plant extract (PPAE). Recordings of the contractile activity of the aortic rings to the extract (1.9-8.6 mg/ml) were done using standard organ bath techniques. Impact on CYP3A4 and CYP2D6 enzyme activities was investigated using human liver and heterologously expressed microsomes. Results: We observed a dose-dependent reduction in systolic, diastolic, MAP and HR. Pre-treatment with atropine (2 mg/kg) and propranolol (1 mg/kg) but not mepyramine (2 mg/kg) significantly (p<0.05) reduced the hypotensive and negative chronotropic activities caused by the extract, while L-NAME (5 mg/kg) completely abolished it. PPAE significantly (p<0.05) relaxed the phenylephrine (10-9-10-4 M) and KCl- induced contractions and displayed moderate inhibition of CYP3A4 enzyme activity with IC50 values of 0.466 ± 0.126 mg/mL and 0.153 ± 0.054 mg/mL, respectively using heterogenously expressed CYP3A4 and human liver microsomes (HLMs) Conclusion: Results suggest dose-dependent hypotensive, bradycardic and vasorelaxant effects of PPAE are mediated through Nitric oxide-dependent mechanisms. The impact on CYPs enzyme activities indicate unlikely adverse drug effect when Peperomia pellucida is consumed with other medications reliant on CYP3A4 metabolism. Keywords: Hypertension; Endothelium; Peperomia pellucid; anthraquinones have also been isolated from the plant [4]. Cytochrome P450 (CYP450); Mechanism of action Given the widespread ethno medicinal use of this plant and its Background usage in combination with other medications, it was important to investigate impact on drug metabolising cytochrome P450s (CYPs), Hypertension is a major cardiovascular burden that has been a heme containing superfamily of enzymes [17], Herbal remedies are estimated to cause 7.1 million premature deaths and 45 % of the disease known to affect the dynamics of drug and chemical interactions [18], burden [1]. However, it remains inadequately managed everywhere [2], and of significant concern to drug-drug interactions [19,20]. There is and in spite of the large number of antihypertensive medications, most therefore need for pharmacological validation of this medicinal plant to people in developing countries have poor access to modern health- justify its usage and safety in ethno medicinal treatment, which could care and cannot afford these drugs due to cost. Therefore, they resort greatly benefit populations with poor economic resources. Therefore, to alternative herbal remedies to manage hypertension.Such alternative the aim of the present study was to investigate the blood pressure remeadies include the use of herbs and natural plant products, one lowering effects and possible mechanism of action of an aqueous of the plant used is Peperomia pellucid [3]. Peperomia pellucida is a extract of Peperomia pellucida, and its possible drug-herb interactions common, fleshy annual herb that belongs to the family of Piperaceae. using CYP540 microsomes. It is commonly called shiny bush, pepper elder, man-to-man, rat-ear, Pansit pansitan and is found mainly in the tropics [4-6]. Methods Traditionally, the plant is used as a diuretic and to reduce cholesterol Chemicals and reagents levels in the treatment of hypertension and kidney disorder [4-6]. It All chemicals except those noted below were purchased from is also reported to have anti-inflammatory properties [6] analgesic activity[4,6-8] antipyretic [9] as well as antibacterial [5,7,10] also used in the treatment of abscesses, furuncles, and conjunctivitis [6]. *Corresponding author: Dr. Chukwuemeka R. Nwokocha, Department of Basic The phytochemicals present in the plant are alkaloids, namely, Medical Sciences, University of West Indies, Mona Campus, Kingston 7, Jamaica; Tel: secolignans, tetrahydrofuran lignans, as well as, highly methoxylated +876 589 5445; Fax: +876 977 3823; E-mail: [email protected] dihydronaphthalenone, peperomins A, B, C, and E, sesamin, and Received March 06, 2012; Accepted May 23, 2012; Published May 28, 2012 isoswertisin [11]. Peperomin E shows growth inhibitory effects on Citation: Nwokocha CR, Owu DU, Kinlocke K, Murray J, Delgoda R, et al (2012) the HL-60, MCF-7, and HeLa three cancer cell lines [7,11]. Peperomia Possible Mechanism of Action of the Hypotensive Effect of Peperomia pelluci- pellucida also contains several essential oils, mainly dillapiole, da and Interactions between Human Cytochrome P450 Enzymes. Med Aromat Plants 1:105. doi:10.4172/2167-0412.1000105 β-caryophyllene and carotol that have high larvicidal activities [12- 14]. Other compounds are flavonoids such as acacetin, apigenin, Copyright: © 2012 Nwokocha CR, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits isovitexin and pellucidatin [15,16], phytosterols, namely, campesterol, unrestricted use, distribution, and reproduction in any medium, provided the stigmasterol, and arylproppanoids. Cardiac glycosides, tannins and original author and source are credited. Med Aromat Plants ISSN: 2167-0412 MAP an open access journal Volume 1 • Issue 4 • 1000105 Citation: Nwokocha CR, Owu DU, Kinlocke K, Murray J, Delgoda R, et al (2012) Possible Mechanism of Action of the Hypotensive Effect of Peperomia pellucida and Interactions between Human Cytochrome P450 Enzymes. Med Aromat Plants 1:105. doi:10.4172/2167- 0412.1000105 Page 2 of 5 Sigma-Aldrich (St. Louis, MO). All CYP substrates and metabolites mepyramine (5 mg/kg) or Nω-nitro-L-arginine methyl ester (L-NAME were purchased from Gentest Corporation (Worburn, MA, USA). 5 mg/kg). Each drug was given intravenously and allowed to equilibrate Escherichia coli membranes expressing human CYP2D6 and CYP3A4 for 5 min before a bolus injection of Peperomia pellucida extract (10 co-expressed with CYP reductase were purchased from Cypex Ltd. mg/kg) (which was a representative dose that gave about 40 – 50% of (Dundee, UK), while pooled human liver microsomes were purchased the effects observed) and the corresponding blood pressure and heart from Xenotech (Kansas, USA). rate changes were recorded. Plant material and extraction Preparation of rat aorta rings ThePeperomia pellucida whole plant (1 kg) was collected in January The thoracic aorta were isolated from male rats and after removal 2010 and the species authenticated by the resident botanist (Mr Patrick of superficial fat and connective tissue, cut into rings of about Lewis) at the herbarium of the University of The West Indies (UWI) approximately 3 mm and mounted in 10 ml organ baths containing where a voucher specimen of the plant material has been deposited normal Krebs physiological solution with the following composition (35447). The whole plant was washed, dried, ground into powder and (mM): NaCl, 112; KCl 5; CaCl2 1.8; MgCl2 1, NaHCO3 25; KH2PO4 0.5; soaked in distilled water overnight. It was filtered using Whatman No. NaH2PO4 0.5; Glucose 10; pH 7.4. The bath solution was maintained at 1 filter paper and the filtrate was concentrated under reduced pressure 37ºC and bubbled continuously with a mixture of 95% O2 and 5% CO2. at 45ºC in a rotary vacuum evaporator. The dark brown solid residue The aorta was connected to an isometric force transducer (SS12LA, was stored in a capped container in a refrigerator at -4ºC until ready Biopac Systems Inc, Goleta, CA, USA), connected to a data acquisition for use. All the drugs used were dissolved in distilled water prior to use. unit (Biopac Student Lab MP36 systems) and isometric contraction All solutions were freshly prepared prior to the start of experimental was recorded using the Biopac BSL PRO computer software. A passive procedures. tension of 1 g was applied to the tissue using a movable device. The rings were equilibrated for 90 min while being rinsed every 15 min. Experimental animals During the equilibration period, the rings were challenged with 1 µM Male Sprague Dawley rats, aged 12 weeks and weighing 300-350 phenylephrine and the aorta was relaxed with 10 µM acetylcholine to g were obtained from the Animal House of the Department of Basic ascertain the endothelial integrity. Medical Sciences, UWI, Mona Campus. They were housed in plastic Characterization of the vasorelaxant responses to Peperomia cages under 12 h light/dark cycles at 25 ± 2ºC and fed with standard rat chow and tap water ad libitum. Ethical approval was sought and pellucida obtained from the FMS/UHWI/UWI, Mona Campus
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