CO-ADMINISTRATION OF GOKO HERBAL CLEANSER AND PARACETAMOL: A HERB-DRUG INTERACTION STUDY

Cecilia Nwadiuto Amadi*image,Oghenetega Ruth Edevbie

Department of Experimental Pharmacology and Toxicology, Faculty of Pharmaceutical Sciences, University of Port Harcourt, Rivers State, Nigeria.

ABSTRACT 

Aim: The present study aimed to evaluate the effects of co-administering paracetamol and Goko cleanserâon the liver. 

Method: Twenty rats divided into four different groups were used for this Study. Group one (control group) received distilled water only; Group two received paracetamol only (100mg/kg), Group three received Goko cleanserâonly (2 mL/kg) and Group four received paracetamol and Goko cleanserâconcomitantly for 3 days. Blood and liver samples were analyzed for biochemical and histopathological parameters respectively. 

Results: Data obtained revealed a significant decrease (p<0.05) in the levels of the liver enzymes (ALT, AST, and GGT) in the Goko cleanserâ-only group compared to control. A significant decrease in AST was also observed in paracetamol + Goko cleanserâgroup. No significant change in ALT, ALP and GGT levels in the paracetamol + Goko cleanserâgroups was observed. Results of histology examination showed normal liver tissues with normal hepatocytes, sinusoids and central vein for both treated and control groups. 

Conclusion: The significant reductions in the liver enzymes by observed in the Goko cleanserâ-only group indicated potential hepato-protective effects which could be due to the presence of phenolic compounds which acted as antioxidants. The short-term co-administration of paracetamol and Goko cleanserâproduced no liver toxicity. However, this is subject to further studies using pharmacokinetic analysis and also, a long-term study is strongly advocated. 

Keywords: Goko cleanser, hepatotoxicity, herb-drug interaction, paracetamol.

 

INTRODUCTION

 

Phytochemicals in foods and herbs have been proven to be beneficial to human health1,2,3. However, the heightened use of herbal medicines in recent times has come with growing concerns associated with the safety and toxicity profile4. Phytochemical constituents can modulate the efficacy and toxic effects of drugs by influencing their metabolism and absorption by interacting with drug transporters and metabolizing enzyme2,5. Co-morbidities of certain diseases have favoured polypharmacy thence posing an increased risk of drug interaction6. This has resulted in the rise in the utilisation of herbal medicines in combination with conventional drugs7. The concurrent intake of herbal and orthodox medicines raises concerns about herb-drug interactions and potential patient safety owing to unclear pharmacokinetic and pharmacodynamic mechanisms of these herbal medicines8. Certain herbal and dietary supplements may produce potentially dangerous herb-drug interaction as they can alter the pharmacokinetics and/or pharmacodynamics of certain medications9. This could be due the presence of adulterants or phytochemicals which could interfere with these processes2,10.  Such herb-drug interactions may be beneficial or detrimental. Interference (induction/inhibition) with CYP isozymes and phase II metabolizing enzymes or drug transporters by phytochemicals is the major mechanisms of drug/herb/nutrient interactions2.

Acetaminophen-induced hepatotoxicity is due to the decreased levels of glutathione due to conjugation with NAPQI (N-acetyl-p-benzoquinone imine)11 a metabolite of acetaminophen metabolism via cytochrome P450 enzyme in the liver12. However, the concentration of NAPQI produced at therapeutic doses are quite little to result in an adverse reaction but when overdosed, it’s concentration increases which results in the depletion of glutathione resulting in hepatotoxicity and necrosis11,12,13. In view of this, an overdose of acetaminophen results to hepatotoxicity amongst other side effects14. 

Goko cleanserÒisa herbal mixture claimed to be effective for various kinds of disease conditions such as, diabetes, infertility, blood clots, hepatitis, erectly dysfunction. Its contents include: Vernonia amygdalina, Cajanuscajan, Zingiberofficinale, Allium sativum, Saccharum officinarum and caramel15. A recent study has identified nephrotoxic effect of Goko cleanser in wistar rats15,16.  Current study aims to probe the effects of a co-administration of Goko cleanserâ and paracetamol on the histology and biochemical parameters of the liver.

 

MATERIALS AND METHODS

 

Chemicals and Reagents

Distilled water, methylated spirit (JHD, China), Xylene, Diethyl ether(JHD Guagdong Guanghua Sci-Tech. co. Ltd. Shantou, Guandong, China), 10% formalin, Goko cleanserâ (Kayfahd herbaceuticals), Paracetamol 500mg tablets BP (Panadolâ from Glaxo Smith Kline). All chemicals and reagents used in this study were of analytical grade.

Source of herbal formulation 

The herbal formulation under study was obtained in August 2020, from Choba open market, Port Harcourt, Rivers State. The herbal formulation was manufactured by Kayfahd herbaceuticals. Exclusively for: Purity biz.com FCT Abuja, Nigeria.

Animals

A previous experimental design described by Ewing et al.,14 was used for this study with little modifications. Twenty (20) healthy adult male rats (152.6 ± 20.6 g) were used for the experiment. The animals were kept in the animal house of the Department of Experimental Pharmacology and Toxicology, Faculty of Pharmaceutical Sciences, University of Port Harcourt, Nigeria for a period of two weeks and fed with standard feed (Broiler finisher- Guinea feeds) ad libitum  with free access to water before experiment. Animal ethics and proper handling methods following during experiments. 

Animal experiment

The dosage for Goko cleanserâ herbal mixture was chosen using its prescription label. Animals were randomly assigned to four groups (1-4) of five animals each. Group I received 0.3mL of distilled water orally daily for 3 days (control group). Group II animals received oral dose of paracetamol (100 mg/kg daily for 3 days). Dose of paracetamol was chosen based on results of LD50 previously determined by Ewing et al14. Animals in group III received oral dose of Goko cleanserâ (2mL/kg daily for 3 days). Group IV animals received both paracetamol and Gokocleanserâ orally at a dosage of 100 mg/kg and 2 mL/kg respectively concurrently daily for 3 days. A conventional/therapeutic usage of paracetamol and Goko cleanserâ was mimicked with this experimental setup. Animals were then fasted overnight on the third day of treatment and sacrificed using ether anaesthesia on the fourth day.

Blood sampling and biochemical analysis

Samples of blood were collected via cardiac puncture and kept for 6 hours at a temperature of 4°C.  Prior to biochemical analysis, blood samples were centrifuged at 3000 rpm for 10 minutes. Liver function was probed with commercial diagnostic kits (Randox laboratory kit, England) using serum levels of Glutamyl transferase (GGT), Alkaline phosphatase (ALP), Alanine amino transferase (ALT) and Aspartate amino transferase (AST).

Histopathological Examination

Liver sections were fixed in 10 % formalin for 6-12 hours. They were processed and examined for histological changes at the college of Health Sciences pathology facility. For light microscopy examination, liver tissues fixed in formalin were dehydrated using increasing grades of alcohol embedded in paraffin. Serial sections were taken and stained with H and E as per standard protocol. Stained sections were morphologically evaluated, and the pictures of the slides were taken for comparison.

Ethical issues

The study protocol was designed in line with the ethical principles of the International Committees for the Protection of Animal Rights Laboratory and approved by the research ethics Committee of the University of Port Harcourt, Rivers State, Nigeria.

Statistical analysis 

Statistical analysis involved use of the Microsoft Excel.  Results were expressed as the Mean ± SD. Analysis were done using one-way Anova and t-tests. P values below 5% were considered statistically significant (p < 0.05). 

 

RESULTS

 

Biochemical analysis

In this study, enzyme concentrations were all within normal ranges. The rats treated with Goko cleanserâ alone showed a significant (p<0.05) decrease in the concentrations of liver function markers, ALT, AST, and GGT enzymes compared to the control group, paracetamol-only group and Gokoâ-paracetamol group. However, there was a mild increase in the concentration of ALP in the Goko cleanserâ-only and the paracetamol/Goko cleanserâ groups when compared with the control. However, this increase was not statistically significant. Similarly, there was a non-significant increase in ALP, ALT and GGT in the Gokoâ-paracetamol interaction group compared to the control group. These results are shown in Figure 1.

Histopathology Results

Results of the histopathology analysis revealed no apparent distortions in the liver tissues from both control and treated groups. The liver tissues were histologically normal showing normal hepatocytes, sinusoids, central vein and portal triad. 

Effects of Paracetamol-Goko CleanserÒ Interaction on Rat Body Weights

The body weights of the rats were determined and are shown in Table 1. All animals exhibited increased body weights at the end of the experiment with significant difference (*p<0.05) between both control and Goko cleanserâ groups. 

Figure 1: Effect of Paracetamol + Gokoâ interaction on liver enzymes; n=5, values are expressed as mean±SEM. * p<0.05 compared to control group.

DISCUSSION      

 

Drug-herb interaction is a matter of concern especially in the developing countries, as herbal remedies are linked with complications such as liver toxicities with high incidence of mortalities and morbidities4. The clinical presentation could range from asymptomatic cases with distorted liver function tests to more severe liver failure4. Herbal remedies have become popular across the globe in primary healthcare, and some have been erroneously regarded as safe just because they are of natural source5. In addition, these therapeutic products from medicinal plants are claimed to be safe without any toxic health effects, and thus widely utilized as self-medications17. However, there are little or no proven scientific studies on the toxicity and adverse effect of these remedies18. 

Table 1:  Showing the effects of paracetamol and Goko cleanserâ interaction on rat body weights.

Recent studies have reported that certain herbs have the potential to induce drug metabolizing enzymes and transporters19. For example, a herbal preparation containing Kaempferia parviflora have been shown to induce CYP2E1 which is an enzyme that metabolizes paracetamol. Paracetamol (acetaminophen) is metabolized to give the major hepatotoxic product known as N-acetyl-p-benzoquinone imine (NAPQI)20. In view of this, Mekjaruskul and Sripanidkulchai19 investigated the combined use of K. parviflora extract with CYP2E1 paracetamol. Results obtained from that study provided a possible drug-herb interaction between K. parviflora and paracetamol leading to increased metabolism and possible enhanced levels of the metabolite NAPQI19. 

Besides the negative effects of drug-herb interaction, some studies have highlighted beneficial effects. For example, certain herbs have been shown to possess ameliorative effects against some drug-induced toxicity. In a study by Hamid et al., Zingiber zerumbet was documented to have protective activity against paracetamol-induced acute hepatotoxicity in rat model21. Further to this, Curcuma longa extract was shown to protect the liver against carbon tetrachloride-induced liver injury22. In addition, some herbs such as Ginkgo biloba have demonstrated beneficial synergistic effects that enhances the efficiency of haloperidol23.

In the present study, the results obtained from biochemical analysis produced a reduction in the levels of liver enzymes in the Goko cleanserâ-paracetamol group compared to control. These results are in agreement with the data obtained from histopathological examination, which showed normal liver tissues with normal hepatocytes, sinusoids, central vein and portal triad (portal vein, portal artery and bile duct). Goko cleanserâ when administered alone and in combination with paracetamol reduced the concentrations of ALT, AST and GGT significantly (p<0.05) compared to control and paracetamol-only groups. While AST levels were significantly reduced in group that received Goko cleanserâ alone and paracetamol + Goko cleanserâ concurrently, the AST level in the paracetamol- Goko cleanserâ group was higher than the Goko cleanserâ -only group suggestive of an elevation accruing from an interaction between paracetamol and Goko cleanserâ. Furthermore, increased body weights of the animals across all treatment groups were suggestive of the absence of toxicities from the co-administration of paracetamol and Goko cleanserâ.

Figure 2: Photomicrograph (400x magnification) of A). Liver from rats in the control group that received distilled water only; B). Liver tissue of rats that received paracetamol only; C). Liver from rats that received Goko cleanserâ only; D). Liver from rats that received paracetamol + Goko cleanserâ.

Taken together, the significant reductions in the liver enzymes by Goko cleanserâ indicated potential hepato-protective effects; this provides some level of support to the claims made by the manufacturer about Goko cleanserâ being hepato-protective. The potential hepato-protective effect of Goko cleanserâ could be attributable to the presence of flavonoids and other phenolicslike tannins contained in some of its constituent herbs15. Flavonoids and tannins are plant-based antioxidants known to reduce the risk of occurrence of numerous human disease related to oxidative stress24. Oxidative stress is known to play a foremost role in the progress of liver diseases25. Earlier studies have highlighted the potential hepato-protective effects of flavonoids and other plant-based antioxidants in herbs. For instance, a study by Kim et al., demonstrated that extracts of Alnus japonica alleviated the acetaminophen induced hepatic injury in rats26. These findings are in agreement with data obtained from our study.

 

CONCLUSION

 

From the results obtained from this study, no hepatotoxic effects were observed with short-term administration of paracetamol and Goko cleanserâ at therapeutic doses in rats. Therefore, the study hypothesizes that a short-term co-administration of the duo poses no potential health risk, however this is subject to further studies. Specifically, pharmacokinetics studies and long-term co-administration studies are strongly advocated.

 

AUTHOR’S CONTRIBUTION

 

All authors analyzed the data, wrote the manuscript and reviewed it.

 

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