Evaluation of some prescribed and over-the-counter drugs induced hemato- biochemical changes in mice

The present study was performed in the Department of Physiology and Pharmacology, Faculty of Animal science and Veterinary Medicine, PSTU, Bangladesh. Data were collected using a semi-structured questionnaire from 125 people in Babugonj upazilla, Barishal for using drugs without prescription or at prescription. The experiment was conducted to evaluate the effects of prescribed and over-the-counter drugs on hematological (TEC, Hb and DLC) and biochemical (blood glucose) parameters in mice during the period March to April, 2019. 140 male mice were randomly divided into seven groups (A, B, C, D, E, F and G), each consisting of 20 mice. Group A was kept as control. Paracetamol, diclofenac, pantoprazole, ranitidine, diazepam and clonazepam were administered orally to mice of group B, C, D, E, F and G, respectively for 28 days. Paracetamol and ranitidine were used by 100% people followed by peantoprazole (80.80%), diclofenac-60%, diazepam-42.40% and clonazepam-24%, respectively without prescription. Paracetamol were used by 100% people followed by peantoprazole (84%), ranitidine-76%, diclofenac-66.40%, clonazepam-53.60% and diazepam 32.80%, respectively at prescription. Paracetamol and diazepam decreased significantly (p<0.05) total erythrocyte count (TEC) at 28 day. Paracetamol, diclofenac and diazepam decreased significantly (p<0.01 and p<0.05) hemoglobin content (Hb) at 28 day. The Differential leukocyte count (DLC) was not significantly changed at 28 day in mice. Paracetamol, diclofenac, pantoprazole, ranitidine, diazepam and clonazepam increased significantly (p<0.01 and p<0.05) blood glucose level in mice. It may be concluded that paracetamol is one of the most frequently used and self prescribing drugs in Bangladesh.


Introduction
Drug is any substance that inhaled, injected, smoked, consumed, absorbed via a patch on the skin, or dissolved under the tongue causes a temporary physiological/psychological change in the body. In pharmacology, drug is a chemical substance when administered to a living organism, produces a biological effect (Adams, 1995). Now-a-days, there is a great problem that so many people use different drugs, either over-the-counter (OTC) drugs or prescription drug, without any concern of physician/doctor or take overdose/subnormal dose of drug without following the prescription unconsciously. That causes different side effects, toxicity, hazard and idiosyncrasy on body. Paracetamol is a widely used analgesic and antipyretic drugs (Anderson et al., 1998;Vane et al., 1998). Diclofenac sodium is a widely circulated drug, used both in humans and animals for the treatment and management of inflammation, fever and pain associated with disease or injury of domestic livestock and humans (Orinya et al., 2016). Pantoprazole are the drug whose main action is pronounced and long-lasting reduction of gastric acid production (Bigoniya et al., 2011). Ranitidine is a medication which established as a potent inhibitor of gastric acid secretion (Pahwa et al., 2016). Diazepam is used for its myorelaxant, anxiolytic, sedative, and anticonvulsant properties (Crestani et al., 2016). Clonazepam acts as sedative, hypnotic, muscle relaxant and anticonvulsants and are especially useful in treating acute anxiety and transient insomnia (Zorzanelli et al., 2019). To maintain sound health, we should conscious of using drugs. We should follow the prescription and avoiding the overdoses or side effects of drugs. In Bangladesh, there is very few research data of drugs are available. The survey will be build up awareness of people about prescribed and OTC drugs to avoid different side effects, toxicity, hazard and idiosyncrasy on body in Bangladesh. The experiment was conducted to assess the use of different types of prescribed and OTC drugs in Babugonj upazilla of Barishal district at prescription or without prescription. The experiment was also evaluate the effects of prescribed and over-the-counter drugs (paracetamol, diclofenac, pantoprazole, ranitidine, diazepam and clonazepam) on hematological (TEC, Hb and DLC) and biochemical (blood glucose) parameters in mice.

Materials and Methods
The experiment was performed in the laboratory of Department of Physiology and pharmacology, Patuakhali Science and Technology University, Babuganj, Barishal, during a period from 10 March, 2019 to 6 April, 2019. A local level survey of people was conducted in Babugonj upazilla of Barishal district of Bangladesh during January, 2019 to February, 2019. Data (information) were collected by a semi-structured questionnaire from 125 people  for using different drugs (paracetamol, diclofenac, pantoprazole, ranitidine, diazepam and clonazepam) at prescription or without prescription. A total of 140 Swiss Albino male mice were collected from the ICDDR,B, Mohakhali, Dhaka. After procurement, all the mice were kept under close observation in order to acclimatize in the new environmental condition of the experimental Pharmacology Laboratory, Faculty of Animal Science and Veterinary Medicine, Patuakhali Science and Technology University for a period of one week prior to commencement of the experiment. All the mice will be maintained under good housing conditions and provide normal feed. One hundred forty (140) male mice were randomly divided into seven groups (A, B, C, D, E, F and G), each consisting of twenty (20) mice. Group A was kept as control group received only basal feed. Paracetamol, diclofenac, pantoprazole, ranitidine, diazepam and clonazepam were administered orally to the mice of group B, C, D, E, F and G, respectively with basal feed with standard dose for 28 days. The effects of six patent prescribed and OTC drugs [paracetamol (Tab. Fast® 500, @15 mg/kg b.wt), diclofenac (Tab. Clofenec ® 50, @50 mg/kg b.wt), pantoprazole (Tab. Trupan ® 20, @15 mg/kg b.wt), ranitidine (Tab. Neotack ® 150, @10 mg/kg b.wt), diazepam (Tab. Sedil ® , @5 mg/ kg b.wt) and clonazepam (Tab. Epitra ® 2, @)2 mg/ kg b.wt] on hemato-biochemical parameters (TEC, Hb, DLC and blood glucose] were evaluated in Swiss Albino male mice. Blood glucose level were recorded on day 0 (pre treatment) and on 7 th , 14 th , 21 st and 28 th day of experimental period by the help of Glucometer. Hematological parameters (TEC, Hb and DLC) were determined on the 28 th day by the standard method as described by Coffin (1953). Collected data were statistically analyzed by Student's t-test by using the computer statistical package programme of Microsoft Excel.

Effects of prescribed and OTC drugs on hematological parameters in mice
Paracetamol, diclofenac, pantoprazole, ranitidine, diazepam and clonazepam caused changes on hematological parameters (TEC, Hb and DLC) in mice. The results of the paracetamol, diclofenac, pantoprazole, ranitidine, diazepam and clonazepam on hematological parameters were shown in the Table 3.

Total erythrocyte count (TEC, million/cu.mm.)
Paracetamol, diclofenac, pantoprazole, ranitidine, diazepam and clonazepam caused changes on total erythrocyte count (TEC) at 28 th day in mice. Paracetamol and diazepam decreased significantly (p<0.05) TEC at the mice of group B and F, respectively. On the 28 th day TEC was 6.31±0.26, 5.24±0.60, 6.44±0.17, 6.19±0.24, 6.22±0.18, 6.13±0.37 and 6.36±0.32, respectively in groups A, B, C, D, E, F and G. These results are agreeable with the report of Oyedeji et al. (2013). He reported that after administrating of paracetamol in albino mice for 42 days significantly decreased total erythrocyte count (TEC). Likewise, Hossain et al. (2003) reported that paracetamol significantly decreased TEC. Similar response has been reported by Suresha et al. (2012) due to diazepam.

Hemoglobin content (Hb, gm %)
Paracetamol, diclofenac, pantoprazole, ranitidine, diazepam and clonazepam caused changes on hemoglobin content (Hb) at 28 th day in mice. Paracetamol, diclofenac and diazepam decreased significantly (p<0.01 and p<0.05) Hb at the mice of group B, C and F, respectively. On the 28 th day Hb was 10.18±0.22, 9.04±0.40, 11.12±0.28, 10.26±0.23, 10.10±0.40, 9.94±0.45 and 10.16±0.53, respectively in groups A, B, C, D, E, F and G. In confirmatory to the present findings, Ukwueze et al. (2014) observed that prolonged use of paracetamol decreased Hb content significantly. This observation is also in agreement with the earlier reports on paracetamol of Oyedeji et al. (2013) and Hossain et al. (2003). Likewise, Orinya et al. (2016) reported that after administrating diclofenac decreased Hb content in wistar rats. Similar results have been observed in dogs following diazepam administration by Suresha et al. (2012).

Effect of prescribed and OTC drugs on biochemical parameter (blood glucose) in mice
Paracetamol, diclofenac, pantoprazole, ranitidine, diazepam and clonazepam caused significant changes on blood glucose level in mice. The blood glucose level was increased significantly (p<0.01 and p<0.05) after Paracetamol, diclofenac, pantoprazole, ranitidine, diazepam and clonazepam treatment in group B, C, E, F and G, respectively. The results of the prescribed and OTC drugs on blood glucose level in mice were shown in the Table 4. Mean blood glucose level of control group on pre-treatment (day 0) was 4.22±0.16. Mean blood glucose level on the 7 th , 14 th , 21 st and 28 th day were 4.58±0.30, 4.90±0.26, 5.22±0.34 and 5.39±0.33, respectively in group A. Mean blood glucose level before treatment was 4.35±0.17 and after treatment with paracetamol mean blood glucose level on 7 th , 14 th , 21 st and 28 th day were 4.87±0.18, 5.33±0.28, 5.86±0.25 and 6.65±0.40, respectively in group B. Mean blood glucose level before treatment was 4.12±0.13 and after treatment with diclofenac mean blood glucose level on 7 th , 14 th , 21 st and 28 th day were 4.86±0.22, 5.53±0.36, 6.05±0.48 and 6.71±0.66, respectively in group C. Mean blood glucose level before treatment was 4.51±0.31 and after treatment with pantoprazole mean blood glucose level on 7 th , 14 th , 21 st and 28 th day were 4.79±0.34, 5.66±0.53, 5.98±0.48 and 7.87±1.53, respectively in group D. Mean blood glucose level before treatment was 4.33±0.19 and after treatment with ranitidine mean blood glucose level on 7 th , 14 th , 21 st and 28 th day were 4.61±0.29, 5.41±0.10, 6.34±0.59 and 7.26±0.94, respectively in group E. Mean blood glucose level before treatment was 4.50±0.37 and after treatment with diazepam mean blood glucose level on 7 th , 14 th , 21 st and 28 th day were 4.66±0.38, 5.12±0.59, 5.88±0.43 and 6.59±0.76, respectively in group F. Mean blood glucose level before treatment was 5.41±0.67 and after treatment with clonazepam mean blood glucose level on 7 th , 14 th , 21 st and 28 th day were 5.58±0.73, 6.52±0.79, 7.39±0.83 and 7.53±1.10, respectively in group G. In confirmatory to the present findings, Hossain et al. (2003) reported that paracetamol significantly increased blood glucose in mice. Similar response has been reported due to paracetamol (Amin et al., 2004), diclofenac (Ahmad et al., 2012), pantoprazole (Agrawal et al., 2018, ranitidine (Gentile et al., 1986), diazepam (Suresha et al., 2012) and clonazepam (Bougherara and Bouaziz, 2014). .10** The above values represent the mean  standard deviation (SD) ** = Significant at 1 per cent level (p<0.01) * = Significant at 5 per cent level (p<0.05)

Conclusions
Self-medication is one component of self-care, more awareness is needed to foster the level of people's attitudes towards self-medication practices and it is important to consider the manners of drug availability and consumption to decrease unnecessary health risks. The study give some information about the use of some prescribed and OTC drugs.

Conflict of interest
None to declare.