An in Vivo and in Silico Evaluation of the Hepatoprotective Potential of Gynura Procumbens: a Promising Agent for Combating Hepatotoxicity
Tanzia Islam Tithi, Md. Rafat Tahsin,Juhaer Anjum, Tasnuva Sharmin Zaman, Fahima Aktar, Nasiba Binte Bahar, Sabiha Tasnim, Arifa Sultana, Ishrat Jahan, Syeda Sadia Afrin, Tahmina Akter, Priyanka Sen, Fahima Jannat Koly, Md. Selim Reza, Jakir Ahmed Chowdhury, Shaila Kabir, Abu Asad Chowdhury, Md. Shah Amran
Abstract
Introduction
The liver, the most important metabolic organ of the body, performs a wide variety of vital functions. Hepatic cell injury occurs by the activation of reactive oxygen species (ROS) that are generated by carbon tetrachloride (CCl4), xenobiotics, and other toxic substances through cytochrome P450-dependent steps resulting from the covalent bond formation with lipoproteins and nucleic acids. Observing the urgent state of hepatotoxic patients worldwide, different medicinal plants and their properties can be explored to combat such free radical damage to the liver. In vivo and in silico studies were designed and conducted to evaluate the antioxidant and hepatoprotective properties of Gynura procumbens in rats.
Materials and methods
Gynura procumbens leaves were collected and extracted using 70% ethanol. The required chemicals CCl4, standard drug (silymarin), and blood serum analysis kits were stocked. The in vivo tests were performed in 140 healthy Wister albino rats of either sex under well-controlled parameters divided into 14 groups, strictly maintaining Institutional Animal Ethics Committee (IEAC) protocols. For the histopathology study, 10% buffered neutral formalin was used for organ preservation. Later the specimens were studied under a fluorescence microscope. In silico molecular docking and absorption, distribution, metabolism, excretion, and toxicity (ADMET) studies were performed, and the results were analyzed statistically.
Introduction
The liver is a vital and resilient organ in an animal’s body, the primary functions of which are to support metabolism, digestion, detoxification, storing vitamins, minerals, and immunity. However, this organ can be affected by toxins from food supplements, drugs, chemicals, and medicinal plants [1–3]. Carbon tetrachloride (CCl4) is considered quite toxic and xenobiotic to the animal body and causes cell injury by activating reactive oxygen species (ROS). Peroxy trichloromethyl (OOCCl3) and trichloromethyl (CCl3) radicals are usually generated during the cytochrome P450-dependent metabolic steps. These free radicals produce covalent bonds with lipoproteins and nucleic acids that cause extensive cell injury in the liver and other vital animal body organs [4–9]. To date, around fifty million people worldwide are suffering from hepatotoxicity. This hepatotoxicity state is quite alarming in the health sector [1, 10]. Symptoms related to hepatotoxicity may include jaundice, fatigue, abdominal pain, skin rashes, itching, rapid and abnormal weight gain, vomiting or nausea, light-colored stool, and dark urine [11, 12]. To examine liver function for diagnostic purposes, serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT), and alkaline phosphatase (ALP) levels are measured in the blood [13]. Synthetic drugs such as thalidomide, curcumin, ademetionine, entecavir, metadoxine, tenofovir, ondansetron, and resveratrol are available in the pharma arena, which are used to treat liver diseases [14]. However, the toxicity and efficacy of these medicines are still under investigation, and medicinal plants have an excellent opportunity to take their position through genetic modification using different biotechnological studies [15]. Medicinal plants have always significantly impacted medicines and nutrition for different societies in the world. Around 70,000 plant species have been used for diseases as medicine.
Method and materials
Collection and extraction of Gynura procumbens leaf
Gynura procumbens leaves were collected from the medicinal plant garden of the Faculty of Pharmacy, University of Dhaka. After that, the Bangladesh National Herbarium certified the specimen and provided the accession number YM001 for future reference. The collected leaves of Gynura procumbens were then carefully washed and kept for 7 days for shed drying. Next, the dried leaf was kept in an oven at 30°C temperature for 10 days. After that, the dried leaf was powdered coarsely. After that, powdered leaves were extracted using 70% ethanol for twelve days, and the extract was filtered every three days. Later, a rotary evaporator was used to dry the extract using low temperature and pressure. Finally, different pharmacological tests were performed using this crude residue.
Results
A significant elevation of body weight was observed after administering Gynura procumbens extract in CCl4-induced hepatotoxic rats [Fig 1]. In the negative control group, body weight was increased at the final stage. However, carbon tetrachloride treatment prompted a decrease in terminal body weight in the disease control group. Groups treated with three different doses of Gynura procumbens (low, medium, and high) following carbon tetrachloride elevated the gradual pattern of weight gain. However, silymarin treatment showed the opposite scenario. A reversal of CCl4-mediated body weight decline was reported upon treatment with different doses of Gynura procumbens extract. Groups administered only plant extract followed a similar trend as the negative control group, whereas a contrasting scenario was observed in groups given silymarin only.
Discussion
In this study, we assessed the hepatoprotective activity of the ethanolic extract of Gynura procumbens through standard parameters. In body weight determination, CCl4-induced hepatic impairment would limit the body’s capacity to metabolize nutrients, resulting in metabolic imbalance and substantial weight loss. Negative control rats gained terminal weight, demonstrating proper metabolic equilibrium [Fig 1]. Only silymarin-treated groups showed the opposite result, indicating its detrimental effect on healthy rats’ growth rate and body weight. 70% ethanolic extract of G. procumbens corrected CCl4-induced weight loss in a dose-dependent manner, demonstrating the extract’s promise in restoring standard growth rate and body weight in diseased rats. We have found prior studies that G. procumbens ethanolic extract restored body weight in diseased mice [79, 80]. Increases in body weight after treatment with plant extracts were also found in the case of Phytolacca dodecandra [1]. According to a study conducted in Bangladesh, we also found that treatment with G. procumbens ethanolic extract resulted in consistent recovery of sick rats’ body weights [81]. Both the aqueous and 90% ethanolic extracts of G. procumbens efficiently recovered body weights in sick rats in a dose-dependent manner [82].
Conclusion
The current study on G. procumbens in rat models provides new insight into hepatoprotective activity. A decrease in SGPT, SGOT, ALP, creatinine, TC, LDL, triglycerides (TG), SOD, MDA, DNA fragmentation ranges, γ-GT levels, and an increase in HDL level in CCl4-treated rats by both standard drug silymarin and G. procumbens leaf extract demonstrated a potential hepatoprotective activity of the plant extract. From the histopathological study, it is also shown that G. procumbens can reverse the pathological state after treating with CCl4. In molecular docking analysis, several bioactive molecules showed optimistic binding affinity to receptor proteins, and the ADMET study displayed their drug-likeness characters. The results of both in vivo and in silico studies give a ray of hope to consider G. procumbens leaf extract as a possible wellspring of the hepatoprotective drug to be used as an alternative medicine for treating hepatotoxicity. However, further studies are needed to establish the responsible active compounds and their possible mode of action.
Citation: Tithi TI, Tahsin MR, Anjum J, Zaman TS, Aktar F, Bahar NB, et al. (2023) An in vivo and in silico evaluation of the hepatoprotective potential of Gynura procumbens: A promising agent for combating hepatotoxicity. PLoS ONE 18(9): e0291125. https://doi.org/10.1371/journal.pone.0291125
Editor: Ahmed E. Abdel Moneim, Helwan University, EGYPT
Received: October 3, 2022; Accepted: August 22, 2023; Published: September 15, 2023
Copyright: © 2023 Tithi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Data Availability: All relevant data are within the manuscript.
Funding: This work was supported by a financial grant from the Grant for Advanced Research in Education (GARE) from Bangladesh Bureau of Educational Information and Statistics (BANBEIS) under the Ministry of Education, Government of the People’s Republic of Bangladesh, Project ID: LS 20191133 (no. 37.20.0000.004.033.020.2016.1053, dated: 13-10-2019).
Competing interests: The authors have declared that no competing interests exist.
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0291125#abstract0
