International Journal of Pharmaceutical Invention

EFFECT OF VERAPAMIL HYDROCHLORIDE ON ACETYLCHOLINE INDUCED CONTRACTION IN FROG RECTUS ABDOMINIS MUSCLE
K. KARTHIGADEVI*, O. MADHAVI USHARANI, I. SESHA MADHAVI, CH.V.R.TANUJA, S. KAVIMANI Department of Pharmacology, College of Pharmacy, Mother Theresa Post Graduate and Research Institute of Health Sciences (Govt. of Puducherry Institution), Indria Nagar, Gorimedu, Puducherry-605006, India. E-mail: karthi.pharma@yahoo.com Verapamil Hydrochloride is an oral and parenteral calcium-channel blocking (CCB) agent belonging to the phenylalkylamine class. Verapamil Hydrochloride is useful for the treatment of angina, hypertension, and for supraventricular tachyarrhythmias. It is considered a class IV antiarrhythmic agent and Verapamil Hydrochloride is more effective than digoxin for controlling ventricular rate in patients with atrial fibrillation. Skeletal muscle contraction is explained by the excitation-contraction coupling and is dependent on the calcium induced release of calcium from the sarcoplasmic reticulum by Ca2+ release channel due to the previous influx of Ca2+ through Dihydropyridine receptor present on the T-tubule which is due to the conduction of action potential on T-tubule. So, an attempt is made to find out the effect of Verapamil Hydrochloride on acetylcholine induced contraction in Frog Rectus Abdominis Muscle. Maximum response produced by acetylcholine in the absence of the blockers was taken as 100% and against that the % response was calculated. Verapamil Hydrochloride produced significant dose dependant inhibition of acetylcholine induced contraction in CRC. Keywords: Verapamil Hydrochloride, Calcium channel, Frog Rectus Abdominis Muscle, Right Hand Side Shift. INTRODUCTION Verapamil Hydrochloride is an oral and parenteral (CCB) calcium-channel agent belonging class. blocking to the of angina, hypertension, and for

supraventricular tachyarrhythmias. It is considered a class IV antiarrhythmic agent and Verapamil Hydrochloride is more effective than digoxin for controlloing
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phenylalkylamine

Verapamil

Hydrochloride is useful for the treatment

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K. KARTHIGADEVI et al

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than digoxin for controlling ventricular rate in patients with atrial fibrillation. Verapamil Hydrochloride was synthesized in 1962 and in 1981, became the first calcium channel blocker to be approved by the FDA. Verapamil Hydrochloride inhibits the influx of extracellular calcium across the myocardial and vascular

contrast to the situation with the L-type channel antagonists success in developing small molecule antagonists of therapeutic utility for these other channel types has thus far been lacking. The reasons for this are explored and potential new directions are indicated including male fertility, bone growth, immune disorders, cancer and schistosomiasis.[ 2 ] To explore and to investigate for new calcium channel blockers first it was thought to have an extensive knowledge over calcium regulation by different calcium channels and their structures to have a look on binding sites of drugs. The concentration of cytosolic free calcium ([Ca2+]i)is critically important for the control of many essential cellular responses. Changes in [Ca2+] can control specialized functions like excitability, contraction, or exocytosis, due to which neurotransmitter release takes place, while also regulating universal cellular activities such as metabolism and gene expression. In most cells, elevations([Ca2+]i)in arise from [Ca2+] entry via Ca2+ channels in the surface membrane, or Ca2+ discharge from internal stores, or both. Mechanisms that tend to return ([Ca2+]i) toward its low resting value include extrusion of [Ca2+] across the surface membrane or resequestration by internal [Ca2+] stores and
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smooth muscle cell membranes. It exerts its activity at the membrane surface of arterial smooth muscle cells and within conductile and contractile tissue in the myocardium, but the serum calcium levels remain unchanged.[ 1 ] The calcium channel antagonists are a mature group of drugs cardiovascular diseases directed at including

hypertension, angina, peripheral vascular disorders and some arrhythmic conditions. Their sites and mechanism of actions have been well explored over the past two decades and their interactions at the 1subunit of L-type channels (Cav1.1-1.4) have made them valuable molecular tools for channel classification and localization. With the realization that other members of the voltage-gated calcium channel family exist Cav 2.1-2.3 and Cav 3.1-3.3

considerable effort has been directed to drug discovery at these channel types where therapeutic prospects exist for a variety of epilepsy, disorders affective including pain,

disorders, In

by

Ca2+

pump

or

Na+-Ca2+
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neurodegenerative disorders, etc.

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exchangers.[ 3 ]

K. KARTHIGADEVI et al

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MATERIALS AND METHOD A pithed frog was laid on its back on the frog dissecting board. The skin on the abdomen was removed and the rectus abdominis muscle was exposed. The one side of the rectus muscle was dissected from the pelvic girdle to the pectoral girdle. The muscle was then pinned to the cork so as to keep it stretched to its normal length and soaked in frog ringer solution. A thread was sewn through each end and the muscle was mounted in organ bath containing frog ringer solution at room temperature. The concentration

and 0.8 g/ml).The tissue was irrigated for 20 minutes. Maximal response

produced by acetyl choline in the absence of the blocker was taken as 100% and against that the percentage response was calculated.[ 4 ] RESULTS Verapamil Hydrochloride at various test doses (0.4 and 0.8g/ml) produced

significant dose dependent inhibition of acetylcholine induced contractions in

concentration response curves. 640g/ml of acetylcholine was required to produce 100% of response where as in the presence (0.4 of Verapamil Hydrochloride 0.8g/ml), the same

response curve was obtained as described by Kulkarni (2009). The experiment was repeated in each set of preparation in presence of Verapamil Hydrochloride (0.4

and

concentration of acetylcholine produced 92% and 66% of responses respectively.

Fig 1: Effect of Verapamil Hydrochloride on Ach Induced Contractions in Frog Rectus Abdominis Muscle.
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K. KARTHIGADEVI et al

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Fig 2: The graph represents the log dose- response curves of Acetylcholine (100 g/ml) in presence of various concentrations of Verapamil Hydrochloride (0.4 and 0.8g/ml). The progressive shift to the right hand side was noted indicating the antagonistic effect of Verapamil Hydrochloride. DISCUSSION Any muscle contraction is possible only through the cellular calcium signaling and cellular calcium regulation is the excitable cell membranes, as well as proper bone formation.[ 3,5 ] Regulation of Ca2+ is under very tight and dynamic control in all major cell

substantial preoccupation of the cell. Heilbrunn described the calcium is the most ubiquitous intracellular signaling molecule, which serves both physiological and pathologically. It plays an important role in signal transduction pathways, where it acts as a second messenger, in neurotransmitter release from neurons by exocytosis, contraction of all muscle cell types. Many enzymes require calcium

compartments

(cytoplasm,

nucleus,

endoplasmic reticulum, mitochondria). In each compartment, through this the control interplay is of

achieved

transmembrane entry, extrusion systems (channels, exchanger, transporters) and of buffering systems (Ca2+binding protein). It is this role of calcium as an

information-bearing

second

messenger

together with the control processes that make possible the existence of calcium controlling drugs. A D, triumvirate of

ions as a cofactor; those of the bloodclotting cascade being notable examples. Extracellular calcium is also important for maintaining the potential difference across
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hormones-vitamin

parathyroid

hormone and calcitonin- that serve to

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regulate excretion control.[6 ]

calcium and

absorption,

calcium and

Dihydropyridine

Receptor

(DHPR)

bone

deposition

present on the T-tubule which is due to the conduction of action potential on Ttubule. The inhibitory effect of Verapamil Hydrochloride and diltiazem may be due to the blockade of receptor operated calcium channel and voltage dependent calcium channel. Sarcoplasmic reticulum and single channel studies have provided evidence that phenylalkylamine calcium antagonists through the inhibit calcium release reticulum

resorption (remodelling), achieves this THE KEY ROLE OF Ca2+ and Ca2+ CHANNEL IN CELLULAR SIGNALING Ca2+channels allow passage of Ca2+ions into the cytoplasm through a selective pore which is opened in response to depolarization of the cell membrane. The Ca2+ flux creates a net inward,

sarcoplasmic

depolarizing current and the resulting accumulation of Ca2+in the cytoplasm can act as a chemical trigger for secretion of hormones
2+

calcium channel/ryanodine receptor. This action has not been observed with dihydropyridine calcium antagonists.[ 7 ] CONCLUSION The acetylcholine induced contractions in isolated skeletal muscle preparation was effectively antagonised Hydrochloride. by Verapamil

and

neurotransmitters,

contraction of muscle and a variety of other Ca sensitive events. Thus, upon sensing membrane potential changes, Ca2+channels simultaneously generate an electrical signal while directly creating an intracellular chemical messenger. This dual ability is unique among the family of ion channels and allows the Ca channel to play a variety of roles in excitationsecretion coupling. Excitation-Contraction Coupling Skeletal muscle contraction is explained by the excitation-contraction coupling and is dependent on the calcium induced release of calcium from the Sarcoplasmic reticulum by Ca2+ release channel (RyR) due to the previous influx of Ca2+ through
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2+

It also showed dose

dependent antagonistic effect. The log dose response curves of acetylcholine in presence of Verapamil Hydrochloride was shifted to the right hand side indicating the antagonistic effect. The antagonistic effect of Verapamil Hydrochloride

and

excitation-contraction

(phenylalkylamine) may be due to the blocking of both L-type Ca2+channel (DHPR) and Ryanodine Ca2+sensitive channel (RyR) which are involved in the skeletal muscle contractions. REFERENCES 1. http://www.druginfosys.com
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2. TriggleDJ, Antagonists: present and

Calcium Clinical future,

Channel uses past,

edition,

2009,

VallabhPrakashan

Publications 5. http://en.wikipedia.org/wiki/Calcium_ channel_blockers 6. Stefan Channel 2004. 7. Ricardo Zucchi, Effect of gallopamil on excitation-contraction coupling, I McDonough, Pharmacology, Calcium Springer,

Biochem

Pharmacology, 74(1), 2007 June 30, 1-9. 3. Richard W. Tsien, Rogery Y. Tsien, Calcium Osallation, Channels, Annual Stores Review, and Cell

Biology, 6, 1990: 715-60. 4. S.K.Kulkarini, Experimental Handbook Pharmacology, of 3rd

General Pharmacology: The vascular system, 27(5), July 1996, 749-7

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