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Effect of Blatta Orientalis (Q) A Homoeopathic Medicine On An Animal Model of Asthma

EFFECT OF BLATTA ORIENTALIS (Q) A HOMOEOPATHIC MEDICINE ON AN ANIMAL MODEL OF ASTHMA

ABSTRACT

Objective: To study pharmacological action of Blatta orientalis (B. orientalis) a homoeopathic preparation in animal experimental models of respiratory disorders.

Methods: The antiasthmatic activity of B. orientalis (Q) mother tincture (Q) was studied on the acetylcholine (Ach) induced bronchospasm in guinea pigs (in vivo), Ach induced contraction in isolated rat ileum. 

Results: B. orientalis (Q) treated guinea pigs showed significant protection against acetylcholine aerosol-induced bronchospasm and also showed significant dose dependent % protection against Ach induced rat ileum contraction.

Conclusion: B. orientalis (Q) may have selective anti-cholinergic bronchodilator activity B. orientalis (Q) is mainly active in acute asthmatic conditions. 

Key words: - Blatta orientalis, antiasthmatic activity, anticholinergic, spasmolytic. 

INTRODUCTION - Bronchial Asthma is one of the common diseases that affect mankind with diverse manifestations. The prevalence of allergy and asthma has risen in the recent years despite an improvement in the general health of the population. (Ring, et. al., 2001) Various economic studies have identified the causes for increase in the prevalence of upper and lower respiratory tract allergic diseases. Some of the postulated reasons are increasing environmental pollution (Passali, et. al., 1999) and increased predisposition of individuals producing excessive antibody in the asthmatic disorders. (Hansen, et. al., 2004).


Homoeopathy is a 200-year-old therapeutic system that uses small doses of various substances to stimulate auto regulatory and self healing processes. Homoeopathy selects substances by matching a patient’s symptoms with symptoms produced by these substances in healthy individuals. Medicines are prepared by serial dilution and shaking, which proponents claim imprints information into water. Although many conventional physicians find such notions implausible, homeopathy had a prominent place in 19th-century health care and has recently undergone a worldwide revival. (Jonas, et. al., 2003).


Homoeopathic medicine continues to attract attention in medical journals and the media as a popular form of complementary medicine whose proposed mechanism of action seems incompatible with mainstream scientific thought and the research evidence for which remains controversial. (Trevor and Marjori, 2006)


Dynamizations of Blatta orientalis (B. orientalis) (Q) have been found to have a strong in vivo bronchodilator activity, which play a fundamental role in acute asthmatic patient. Therefore, the objective of our study was to evaluate, through an animal-based model, the efficacy of the B. orientalis (Q) remedies, which are usually prescribed by homoeopathists to treat clinical conditions characterized by acute asthmatic manifestations.

MATERIALS AND METHODS
Mother Tincture: Mother tincture (Q) of Blatta orientalis (B. orientalis) 
B. orientalis remedy - Respiratory system: - B. orientalis (Q) is an excellent remedy for Bronchial Asthma in Acute cases, even in status asthmaticus. It acts better in low potencies but in chronic cases it needs higher potencies. Asthma in the form of episodic dry spasmodic persistent cough, breathlessness, followed by scanty muco-purulent discharge and threatened suffocation.  


Choice of Dose (Potencies):- Mother tincture (Q). (Boericke, 1996) 
According to Homeopathic Pharmacopoeia Laboratory Ghaziabad -201002 standards recommended for raw drugs and finished products used in homeopathy 1986 - For Blatta O, the recommended potencies were Mother tincture (Q)-6X. (Homeopathic Pharmacopoeia, 1986)
Dose:- 10-20 drops, thrice daily or more frequent by depending on the severity of the case and dose.


General- Lowest potencies during an attack. Hence from above literature, Blatta orientalis (Q) for the rat, 0.1 mL p.o. and for the guinea pig 0.2 mL p.o. once a day was selected.

Animals: Inbred Wistar rats (150-200 g) and guinea pigs (300-400 g) of either sex housed in standard conditions of temperature (22 ± 2oC), relative humidity (60 ± 5%) and light (12 hours light / dark cycles) were used. They were fed with standard pellet diet and water ad libitum. The study has got approval from Institutional Animal Ethics Committee (IAEC) of R.C.P., Shirpur, India and was in accordance with the guidelines of the committee for the purpose of control and supervision of experiments on Animals (CPCSEA). Registration No. 651/02/C/CPCSEA


Acetylcholine (Ach) induced bronchospasm in guinea pigs (Vogel, 2002; Anil, 2002; Mitra et. al. 1999). Symptoms like dyspnoea, asphyxia, convulsions resembling bronchial asthma in patients can be induced by inhalation of acetylcholine (Ach) or other bronchospasm inducing agents in guinea pigs. The end point pre-convulsive dyspnoea (PCD), asphyxic convulsions was determined from the time of aerosol exposure to the onset of dyspnoea leading to the appearance of asphyctic convulsions i.e. Pre-convulsion time (PCT). Guinea pigs of either sex weighing 300-400 g were exposed to Ach (Loba Chemie Pvt. Ltd. 57608) with constant flow rate 5 mL / min in an aerosol chamber to induced experimental bronchial asthma. This PCT was considered to be T1 value. Two and a half hours later, the guinea pigs were divided in groups like Group I control (Alcohol treated), II B. orientalis (Q) 0.2 mL p.o. and III standard drug (Std.) 2mg / kg of atropine sulfate (Loba Chemie Pvt. Ltd. Art. 1580) treated respectively. These animals were subjected to acetylcholine challenge one and a half hours after receiving the drug and the PCT was noted. This PCT was considered to be T2 value.

Animals which withstood exposure to acetylcholine aerosol for 15 min were considered to be completely protected. As soon as PCD commenced, the animals were removed from the chamber and placed in fresh air. The protection offered by the treatment was calculated by the following formula. 


Percentage protection = 1- T1 / T2 * 100 
Where: T1 is time for PCT before treatment and T2 is the time for PCT after treatment.
Acetylcholine induced contractions on isolated rat ileum (Vogel 2002; Kulkarni, 2005). Overnight fasted rat was sacrificed using cervical dislocation method. Ileum is quickly dissected out washed in Tyrode solution and suspended in organ baths containing Tyrode’s solution (NaCl 9.0, KCl 0.42, CaCl2 0.24, NaHCO3 0.15, Glucose 1.0, Mgcl2, NaH2PO4 gm/ l of distilled water) maintained at 370 C under basal tension 0.5 g and gassed with carbogen. Isotonic contractions were recorded via an isotonic tansduced on polygraph. Thirty minute was allowed for equilibration before the addition of the Ach spasmogen. Ach: - 5 × 10-5 g / mL. When the contraction was reached its maximum at dose response curve (initial spasm) after 10-12 min, the test drug B. orientalis (Q) 0.025 mL was injected in organ bath. Without washing the solution in the organ bath maximum sealing dose of Ach was again inject in it, and response was recorded. After giving washing, sealing dose response of Ach was taken for determining reversibility or irreversibility. Like this, responses of test drug B. orientalis (Q) 0.05 mL, and 0.1mL against Ach contraction were taken till complete inhibition. Percent (%) protection was calculated. 


% protection = Ach max response in absence of test drug - 100 %
Ach max response in presence of test drug - ? 
Calculate % inhibition and plot graph.


Statistical analysis - The results of various studies were expressed as mean ± SEM and analyzed statistically using ANOVA with Tukey-kramer multiple comparison post hock test to find out the level of significance using GraphPad Prism-5. The minimum level of significance was fixed at p < 0.05.

RESULTS
Acetylcholine (Ach) induced bronchospasm in guinea pigs - B. orientalis (Q) significantly prolonged the PCT as compared to control following exposure to acetylcholine aerosols (Fig 1). B. orientalis (Q) showed 87.09 % protection against Ach induced bronchospasm respectively (Table 1). While atropine 2 mg/kg, p.o. was offered significant protection by increasing the mean exposition time in animals exposed to acetylcholine aerosol and showed 88.2 % protection against Ach induced bronchospasm respectively. B. orientalis (Q) showed comparable percent protection against ach induced bronchospasm with atropine. 

Acetylcholine induced contractions on isolated rat ileum. Rat ileum is a known sensitive tissue for studying the effects of anticholinergics. Ach at a dose of 0.4 mL (200ng/mL) was able to produce sealing contraction on isolated rat ileum (8.6 cm taken as 100 %). B. orientalis (Q) showed very low dose dependent antagonistic effect on Ach induced contraction at selling dose expressed in % inhibition (Fig 2 and Table 2). B. orientalis (Q) showed reversible spasmolytic and anticholinergic activity.

DISCUSSION
1. Dyspnoea, asphyxic convulsions like symptoms resembling bronchial asthma in patients can be induced by inhalation of acetylcholine (Ach), histamine or other bronchospasm inducing agents in guinea pigs. Guinea pig is one of the most sensitive animals for asthma and allergic disease. 

2. The first symptom observed was increased breathing frequency followed by forced inspiration and finally convulsions. 

3. The occurrence of these symptoms can be delayed by antagonistic drug. (Vogel, 2002) The end point pre-convulsive dyspnoea (PCD) was determined from the time of aerosol exposure to the onset of dyspnoea leading to the appearance of asphyxiatic convulsions i.e. Pre-convulsion time (PCT). (Mitra, et. al., 1999) Prolongation of PCT indicates spasmolytic and anticholinergic activity of B. orientalis (Q). 

4. It is comparable with standard atropine drugs. 

5. For more conformation of anticholinergic and spasmolytic activity, in-vitro rat ileum activity was done. B. orientalis (Q) was showed reversible anticholinergic activity. 

6. Ach like neurotransmitter induces brochoconstriction. Ach is an important mediator of immediate, allergic and inflammatory reactions. It causes brochoconstriction by activating on M3 (Gq IP3/DAG pathway) receptors. M3 receptors are present in the smooth muscles like GIT, bronchi, uterus. Smooth muscles in most organs are contracted (Rang, et. al. 2003). 

7. Contractility, tone and peristalsis of the GIT and bronchi are increased by Ach action on M3 receptors. In this method, drugs are tested for their capability of inhibiting ileum contraction induced by Ach. It was used to detect anti-cholinergic properties of the test compound. 

8. The reduction in peristalsis and contraction of ileum indicated the reversible anticholinergic (may be M3 antagonist), spasmolytic activity of B. orientalis (Q). 

9. From the above experimentation we can safely conclude that Blatta orientalis (Q) has ANTI-CHOLINERGIC and SPASMOLYTIC activity and it can be used specifically in ACUTE (STATUS ASTHMATICUS) and INTRINSIC (idiosyncratic) ASTHMA. 

10. Further studies may help to establish the anticholinergic activity in other tissues, other modes of action and also identify the active principle responsible for the action.

Treatment % Protection against Ach
Test B. orientalis (Q) 87.09%
Std. Atropine 88.2%
Table 2 Percent inhibition against Ach induced rat ileum.
Dose of B. orientalis (Q) % inhibition against Ach induced contraction of ileum
0.025 mL 66.28 %
0.05 mL 83.73 %
0.1 mL 94.19 %

 

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