Liver disease, a multi-etiological lethal complication, has steadily become one of the significant threats to public health. Liver disease is a substantial global health burden and a leading cause of morbidity and mortality worldwide. It develops with non significant clinical signs or symptoms. Hepato complications are observed in the immune system, body homeostasis, bile formation, protooncogene, and liver-related difficulty 1. On the progression of liver disease, there is a significant elevation of recruited active hepatic macrophages that mark the role of innate immune cells 2. Inflammatory liver illnesses are characterised by recruitments of neutrophil, eosinophil, glutamate, proinflammatory interleukins, nitric oxide, and macrophagic Kupffer cells (kcs), which later cause acute cell fibrosis and uncertain death 3. Various toxins such as drug intoxication (e.g., acetaminophen overdose), viral or autoimmune hepatitis, Wilson's disease, and Budd-Chiari syndrome can all lead to acute liver failure (ALF), including loss of standard histological architecture, decreased cell mass, and decreased blood flow 4. As a result, functional liver capacity gets damaged 5. All negative impact on the liver was seen due to copious consumption of alcohol, abnormal accumulation of adipose tissue, and external insults followed by an imbalance of defensive factors such as Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Gama glutamyl transferase (GGT) or alkaline phosphatase (ALP) 6. Levels of serum AST or ALT are usually increased up to 1.5- to 4-fold but rarely exceed five times the upper limit of normal in the setting of non-alcoholic fatty liver disease (NAFLD). GGT and ALP levels may also be elevated. However, serum prothrombin time, bilirubin level, and serum albumin level are average, except in patients with NAFLD-associated cirrhosis 7. According to World Health Organization 2018, alcohol consumption is responsible for three million deaths globally yearly, accounting for roughly 14% of overall mortality among persons aged 20 to 40 8. Approximately 2 million people die yearly from liver disease, one million from cirrhosis complications, and one million from viral hepatitis and hepatocellular cancer (HCC) 9. Non-alcoholic fatty liver disease (NAFLD) is a growing health concern, with an estimated global prevalence of 25%. According to a recent modelling approach, NAFLD cases in the United States would increase from 83 million in 2015, or about 25% of the population, to 100 million in 2030, or more than 33% of the population 10.

Treatment options for common liver diseases are limited and under clinical trials. Conventional or synthetic drug therapy used in treating liver diseases may lack efficacy and sometimes have serious adverse effects 11. The effectiveness of treatments such as corticosteroids and interferons is inconsistent, carries the risk of adverse events such as ecchymosis, thinning of hair, acne, mild hirsutism, flu-like symptoms, haematological toxicity, and is often too costly 12. Other anti-TB drugs such as Isoniazid, rifampicin, and pyrazinamide can also bring significant adverse effects such as neurological disorders, skin reactions, and gastrointestinal and hepatotoxicity 13. It was reported that interferons could cause adverse effects such as headache, skin eruption, and influenza-like symptoms during treatment with some hepatoprotective drugs such as Bicyclol. Lamivudine can induce mitochondrial toxicity 14.

Therefore, in the absence of a reliable liver protective drug in modern medicine and the view of severe undesirable side effects of synthetic drugs, there is a growing focus on following systematic research methodology to evaluate the scientific basis for traditional herbal medicines for the treatment of liver disorders that are claimed to possess hepatoprotective activity. As a result, they are gaining importance in India and worldwide for their long-lasting curative effect, easy availability, natural healing, and fewer side effects 15.

One of the plants, Sicyos edulis, is believed to cure liver disease. The world's "Plant list" data also supports this proposed investigation due to their ethnopharmacological evidence and traditional therapeutical utilisation. This plant is also used as a vegetable and fruit in most habitats in north-eastern India and the mainland 16. It is a herbaceous perennial climbing plant with tendrils and tuberous roots, cultivated since pre-Colombian times in Mexico. It is reported that the plant Sicyos edulis belongs to the Cucurbitaceae family and is used for treating dysentery, various uterus problems and inflammations, burning sensation, fatigue, various liver disorders, Jaundice, etc. So, Sicyos edulis was selected and aimed to establish the action and potential against the laboratory parameters such as aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and total protein for rapid onset hepatic disease 17.

The present study aimed to carry out the hepatoprotective activity for Sicyos edulis leaf on Wister albino rats, followed by a paracetamol-induced model and a Methotrexate model. The collection of Sicyos edulis leaves was executed during September from the Duga region, East Sikkim, in September. The study was initiated with the authentication of Sicyos edulis, belonging to the family Cucurbitaceae. The leaves previously kept for drying were coarsely powdered and subjected to Soxhlet extraction to facilitate the extraction process. The solvent that was used for the extraction was Methanol. Based on the literature review, methanol was an ideal solvent for extracting the flavonoid and polyphenol compound, the main phytoconstituent of interest for the expected pharmacological activity in future work 24. The 200gm of coarsely powdered plant material gave a yield of 17.4%. The previous literature review confirmed alkaloids, phenols, flavonoids, saponins, proteins, carbohydrates, and tannins in the Sicyos edulis leaf extract 25. The extract's phytochemical estimation revealed the presence of flavonoids, carbohydrates, reducing sugars, and tannins in the section. However, the presence of alkaloids, saponins, and proteins was not determined through the phytochemical estimation, which can be due to solvent selection or the variety of the fruit. The source and location from which the plant was collected can also impact the phytoconstituent in the particular plant. The number of phytochemicals in the extract gives the various pharmacological activities an advantage. Plants can be used to treat various diseases as they contain many phytochemical constituents, such as phenolic and flavonoid compounds, which exhibit a variety of pharmacological activities. These plants can be used in several ways to promote and maintain good health in an individual. Present research includes the presence of carbohydrates, in which restriction plays a significant role in the modulation of lipid metabolism by stimulating lipase enzymes 26. Furthermore, carbohydrates improve NAFLD compared with low-fat diets related to liver diseases. So, the presence of carbohydrates might influence MESEL on liver protection 26. Reducing sugar in the current investigation indicates the pharmacodynamic property of Sicyos edulis; here, pharmacodynamic properties refer to MESEL, affinity towards receptors, protein of interest, and genetical expression 27. The extract has phenolic and flavonoid compound, which is known to exhibit antioxidant property and helps reduce the various reactive oxygen species formation. ROS is said to be involved in the pathogenesis of different diseases, and the case of liver disease, ROS plays a vital role in its development 28. The current investigation on Sicyos edulis reports the presence of tannins. The part of tannin was also established in earlier hepatoprotective studies. This includes significantly reducing serologic enzymes such as ALT, AST, ALP, and T.P. 29. Therefore, it is speculated that the extract of Sicyos edulis leaf can help alleviate the development of liver disease by inhibiting the production of ROS and other factors involved in the progression of the disease. The antioxidant assay was performed by using DPPH. The % radical scavenging activity of the plant extract was 3.6%.

Therefore, in the proposed research, the hepatoprotective activity of the methanolic extract of Sicyos edulis leaf will be evaluated on Paracetamol and induced liver disease. This proposed investigation will build a bridge between chayote's pharmacodynamics and pharmacokinetics in pre-clinical studies on experimental laboratory animals.

The preliminary study of hepatoprotective activity was carried out for Sicyos edulis leaf, followed by an in vivo paracetamol model. Paracetamol was administered with a dose of 1gm/kg body weight subjected to develop hepatic damage in Wister albino rats.

After administering Paracetamol orally, its absorption was delayed due to food entrapment in the stomach and extended the peak plasma concentration with overdose. The metabolism occurs in the liver in phases: Phase I, II and III. Usually, 90% of the drug is metabolised in the Phase II reaction and excreted through urine 30. Still, the remaining 10% of the drug metabolised in Phase I response and produces a highly reactive metabolite called as N-Acetyl P-benzoquinone imine molecule, which impacts glutathione and increases free radicals such as hydrogen oxide, hydrogen superoxide, and nitric oxide and suppresses the oxidation reaction at mitochondria to suppress the ATP synthesis that will further cause damage to DNA of the mitochondrial cell, protein unfolds, and cell necrosis. It also releases mitochondrial contents such as apoptosis, karyolysis, vacuolisation, and inflammation 31. These inflammatory cascades elevate injury to the liver and its enzymatic functions at the cellular and molecular levels. In our present investigation, Paracetamol causes severe liver cell damage, as reported on the 8^th day of laboratory parameters. There was an elevated level of AST in all the animals except the negative control highlighted in Table 1. An increase in AST level is often a sign of liver disease. An increased AST level is observed in serum due to scarring of the liver, permanent damage of liver tissue, myocardial infarction, hemochromatosis, liver ischemia, uncontrolled growth of liver cancer, and consumption of toxic drugs to the liver, primarily physically dependent drugs 32. An ALT blood test aims to help evaluate the liver's health. If cells in the liver are damaged, it can cause ALT to leak into the blood, so an ALT blood test can help find liver issues; risk factors associated with rising ALT could be the intake of more alcohol, Family history, diabetes, Obesity, improvement in ALT levels have shown that the standard Silymarin drug exhibited its hepatoprotective activity 33. ALP is an enzyme that's found throughout the body. ALP blood tests measure the level of ALP in the blood that comes from the liver and bones, and it's one of the tests included in a comprehensive metabolic panel. High levels of ALP in the blood may indicate liver disease or certain bone disorders. ALP is often considered a liver enzyme because its primarily found in the liver. However, it also exists in the following places: the Bile duct, Bones, kidneys, Intestines, and Placenta in pregnant people; abnormal levels of ALP in the blood can reflect damage to tissues or disruption of normal bodily processes 34. The TP test measures the total amount of two proteins found in the fluid portion of blood. These are albumin and globulin. Proteins are essential components of all cells and tissues. Albumin helps to keep fluid from leaking out of our blood vessels. It also transports chemicals in the blood. Globulins are an essential part of the immune system. Low total protein may indicate bleeding, liver disorder, malnutrition, and inflammatory conditions 35.

In our current investigation, statistics reveal that silymarin can restore elevated AST and ALT and control the protein breakdown highlighted in Table 2. The recovery of biochemical parameters supported by Pharmacokinetic studies with silybin-phosphatidylcholine complex has shown an increase in the oral bioavailability of silybin in healthy human subjects, probably by a facilitatory role of drug complex on the passage of the drug across the gastrointestinal tract. Silymarin showed histopathological evidence of hepatoprotection by preventing hepatic cell necrosis or regeneration 36. Silybin di- hemi succinate, a soluble form of silymarin flavonoid, has protected rats against liver glutathione depletion and lipid peroxidation induced by acute acetaminophen hepatotoxicity potential benefits of silymarin as an antidote. Scientists discovered that silymarin treatment normalised the elevated biochemical parameters of the liver and serum caused by acetaminophen by stabilising the plasma membrane in vitro studies on rat hepatocytes 37.

The studied plant, Sicyos edulis, comprises rich secondary metabolites such as carbohydrates, flavonoids, reducing sugar, and tannins, which are also shown in Table 1; this is attributed to the hepatoprotective property of plant in paracetamol-induced liver injury at both doses 100, 200mg/kg as low, and high doses respectively for reducing elevated ALT, AST, ALP, and inhibiting proinflammatory protein breakdown. Furthermore, several pharmacological studies concluded that the Sicyos edulis plant helps the body from free radical damage and slows down the progression of hepatic tissue impairment caused by inducing the Paracetamol model. A previous study reported the ethanolic extract of Sicyos edulis roots against Paracetamol-induced liver damage in rats at 300 and 600mg/kg. It was revealed that the elevation of liver serum biomarkers AST, ALP, and ALT could reverse the hepatic damage to normal and increase the T.P., which supports the hepatoprotective activity of the Sicyos edulis plant 17. However, we have challenged the plant Sicyos edulis leaf with methanolic extract and different fractions of Sicyos edulis (100 and 200mg/kg). It was found to reduce the levels of liver serum biomarkers effectively. Furthermore, our study recorded the higher relative potency at a high dose of Sicyos edulis compared to silymarin.

Methotrexate is well-known to cause serum aminotransferase elevations and long-term therapy has been linked to the development of fatty liver disease, fibrosis and even cirrhosis. In addition, with high-dose intravenous Methotrexate, serum markers will be imbalanced, resulting in indications of liver damage by depletion of purine and pyrimidine synthesis in the liver cells 38.

The current acute model also supports the Methotrexate to induce hepatic toxicity even at a single dose, which was progressively increased till the 7^th day with clear signs of black, tarry stools, itching, rash, reddening of the skin, swelling of the eyelids, hands, feet or lower legs.

With the repeated dose of Silymarin, chayote methanolic extract, significant recovery was observed in clinical signs, symptoms, and evidence yields from laboratory parameters restoration such as AST, ALT, ALP, and total protein. In addition, Silymarin shows reproducible results against methotrexate-induced liver injury at 50mg/kg compared to complicated hepatic animals, and recovery under silymarin was well visible.

This made Sicyos edulis a permissible candidate for further hepatotoxicity studies, including special examinations such as carcinogenicity, mutagenicity, and reproductive, and Tier I studies for long-existing studies.

The Cucurbitaceae family of chayote is widely accepted as a vegetable in the Himalayan region and observed nationwide. Ethnobotany includes cardioprotective, anti-epileptic, anti-obesity, anti-microbial, and anti-diabetic. Chayote is a herbaceous climbing tree whose different parts, such as fruits, leaves, roots and stem, have been researched extensively for its medicinal benefits based on definitive evidence. The current study gave several phytochemicals such as carbohydrates, reducing sugar, polyphenols, flavonoids, and tannins. In the present investigation, our findings provided evidence to support Sicyos eduli's protective effect against Paracetamol and methotrexate-induced hepatotoxicity. In addition, the results showed that the MESEL protected hepatocyte cells against injury by Paracetamol and Methotrexate. However, MESEL at a dose of 200mg/kg has shown a more significant effect on the development of Paracetamol and methotrexate-induced liver injury, which can facilitate the elevated levels of serum biomarkers AST, ALT, ALP and T.P. Our study also added therapeutic and nutritional value in Indian kitchens for food technology.

Thus, the current study confirms the methanolic extract of Sicyos edulis leaf protective action in rats against the Paracetamol and methotrexate model. Furthermore, the quote was very promising, as evidenced by the reversal of the altered values after administration, most likely by promoting hepatocyte regeneration, which restores integrity.