Cytotoxic Effect of the Crude Alcoholic Extract of the Fruits of Citrullus Colocynthis on Human Hepatocyte Carcinoma (Hep-G2).

The Citrullus colocynthis L is a perennial herbaceous plant belonging to the family Cucurbitaceae. Several pharmacological investigations have been performed based on the medicinal application of Citrullus colocynthis. The anticancer and antidiabetic activities of fruit and seed extracts of Citrullus colocynthis have been studied. Newly developed anticancer/antitumor medications appear to have been developed based on the extracted chemicals from Citrullus colocynthis due to the high contents of cucurbitacins. The present study aimed to identify the cytotoxic effect of the crude alcoholic extract of plants of Citrullus colocynthis on the growth of human hepatocyte carcinoma (Hep-G2). The results of the chemical (preliminary) examination of the extract indicated that the fruits contain most of the secondary metabolites including Flavonoids, Tannins, Sapiens, Resins, Amino acids, Glycosides, Terpenes, Alkaloids, and Flavonoids. The toxicological effect of the crude extract was investigated by using six half dilutions concentrations of 20,10,5,2.5,1.25, and 0.625 µg/m at three exposure periods of 24,48, and 72 h using MTT testing. The toxicological effect of the extract appeared for all six concentrations in the Hep-G2 cell line. The highest concentration of 20 µg/ml had the highest percentage inhibition rate with a significant difference (P≤0.01) and reached 93.36 ±1.61 after 72 h of exposure. While the lowest concentration of 0.625 µg/ml was recorded rate of inhibition of 23.36 ± 2.34 after 24 h of exposure. The findings of the present study concluded that the Citrullus colocynthis is one of the most promising medicinal plants which effectively treats cancer through its inhibitory effect and fatal toxicity on cancer cells.

The Unique Substance, Lidocaine and Biological Activity of the Dioscorea Species for Potential Application as a Cancer Treatment, Natural Pesticide and Product.

The six Dioscorea species, D. brevipetiolata, D. bulbifera, D. depauperata (Dd), D. glabra (Dg), D. pyrifolia and D. hamiltonii were analyzed for phytochemicals, toxicity in PBMCs, and biological activity in two cancer cell lines by MTT and comet assays, and pesticide efficiency. Via GC-MS, lidocaine was found to be the predominant compound in two of the studied species. To confirm the systematics, lidocaine was also found in lower amounts in 11 species. The MTT assay showed no toxicity in all six of the studied species. The comet assay showed the key result that the ethanol extracts of Dd and Dg violently broke DNA into pieces. Biological activity of these two species' extracts showed toxicity on HepG2 and no effects on HCT-116. The water extracts of Dd and Dg, applied to Brassica chinensis showed high efficiency as a bioprotectant. In summary, lidocaine seems to be the predominant identifying compound of the genus Dioscorea in Thailand, which is useful in systematics. At least the two species, Dd and Dg, may be used for human hepatocyte cancer treatment and as an alternative pesticide for economically important vegetables. Dioscorea species containing lidocaine or extracted lidocaine have promise for natural product creation.

Previously published ethno-pharmacological reports reveal the potentiality of plants and plant-derived products used as traditional home remedies by Bangladeshi COVID-19 patients to combat SARS-CoV-2.

Several plants have traditionally been used since antiquity to treat various gastroenteritis and respiratory symptoms similar to COVID-19 outcomes. The common symptoms of COVID-19 include fever or chills, cold, cough, flu, headache, diarrhoea, tiredness/fatigue, sore throat, loss of taste or smell, asthma, shortness of breath, or difficulty breathing, etc. This study aims to find out the plants and plant-derived products which are being used by the COVID-19 infected patients in Bangladesh and how those plants are being used for the management of COVID-19 symptoms. In this study, online and partially in-person survey interviews were carried out among Bangladeshi respondents. We selected Bangladeshi COVID-19 patients who were detected Coronavirus positive (+) by RT-PCR nucleic acid test and later recovered. Furthermore, identified plant species from the surveys were thoroughly investigated for safety and efficacy based on the previous ethnomedicinal usage reports. Based on the published data, they were also reviewed for their significant potentialities as antiviral, anti-inflammatory, and immunomodulatory agents. We explored comprehensive information about a total of 26 plant species, belonging to 23 genera and 17 different botanical families, used in COVID-19 treatment as home remedies by the respondents. Most of the plants and plant-derived products were collected directly from the local marketplace. According to our survey results, greatly top 5 cited plant species measured as per the highest RFC value are Camellia sinensis (1.0) > Allium sativum (0.984) > Azadirachta indica (0.966) > Zingiber officinale (0.966) > Syzygium aromaticum (0.943). Previously published ethnomedicinal usage reports, antiviral, anti-inflammatory, and immunomodulatory activity of the concerned plant species also support our results. Thus, the survey and review analysis simultaneously reveals that these reported plants and plant-derived products might be promising candidates for the treatment of COVID-19. Moreover, this study clarifies the reported plants for their safety during COVID-19 management and thereby supporting them to include in any future pre-clinical and clinical investigation for developing herbal COVID-19 therapeutics.

In Vitro Metabolism of 25B-NBF, 2-(4-Bromo-2,5-Dimethoxyphenyl)-N-(2-Fluorobenzyl)ethanamine, in Human Hepatocytes Using Liquid Chromatography⁻Mass Spectrometry.

25B-NBF, 2-(4-bromo-2,5-dimethoxyphenyl)-N-(2-fluorobenzyl)ethanamine, is a new psychoactive substance classified as a phenethylamine. It is a potent agonist of the 5-hydroxytryptamine receptor, but little is known about its metabolism and elimination properties since it was discovered. To aid 25B-NBF abuse screening, the metabolic characteristics of 25B-NBF were investigated in human hepatocytes and human cDNA-expressed cytochrome P450 (CYP) and UDP-glucuronosyltransferase (UGT) enzymes using liquid chromatography⁻high resolution mass spectrometry. At a hepatic extraction ratio of 0.80, 25B-NBF was extensively metabolized into 33 metabolites via hydroxylation, O-demethylation, bis-O-demethylation, N-debenzylation, glucuronidation, sulfation, and acetylation after incubation with pooled human hepatocytes. The metabolism of 25B-NBF was catalyzed by CYP1A1, CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP2J2, CYP3A4, and UGT2B7 enzymes. Based on these results, it is necessary to develop a bioanalytical method for the determination of not only 25B-NBF but also its metabolites in biological samples for the screening of 25B-NBF abuse.

A feasibility study of the toxic responses of human induced pluripotent stem cell-derived hepatocytes to phytochemicals.

Herbal medicines have been increasingly used in the last three decades. Despite their popularity, safety issues with herbal products need to be addressed. We performed a feasibility study of the toxic responses of human induced pluripotent stem cell-derived hepatocytes (iHep cells) to phytochemicals in comparison with hepatoblasoma-derived HepG2 cells and long-term human hepatocytes (LTHHs). The iHep cells expressed typical hepatocyte markers cytochrome P450 3A4 (CYP3A4), hepatocyte nuclear factor 4α, and albumin despite the expression of immature markers α-fetoprotein and cytokeratin 19. We studied the responses of iHep cells to phytochemicals saikosaponin D, triptolide, deoxycalyciphylline B, and monocrotaline with different mode of toxicity employing MTS and lactate dehydrogenase (LDH) assays. Saikosaponin D and triptolide caused dose-dependent cytotoxicity in the iHep cells, which were more sensitive than LTHHs and HepG2 cells. Saikosaponin D-induced cytotoxicity tightly correlated with increased LDH leakage in the iHep cells. Although deoxycalyciphylline B did not exhibit toxic effect on the iHep and HepG2 cells when compared with LTHHs, it decreased CYP3A7 expression in the iHep cells and increased CYP1A2 expression in HepG2 cells. We hereby show the feasibility of using iHep cells to detect toxic effects of phytochemicals.

How useful are clinical liver function tests in in vitro human hepatotoxicity assays?

In preclinical hepatotoxicity testing cell based assays are frequently employed. However, prediction of clinical drug induced liver injury (DILI) remains a major challenge. Here we examined the usefulness of frequently employed markers of hepatocellular injury in cultures of primary human hepatocytes (PHH) in response to treatment with either paracetamol, rifampicin, petadolex and/or amiodarone. The changes in the metabolic competency (urea and albumin) and cellular injury (AST, ALT, ALP, LDH, γGT and succinate dehydrogenase) were determined at therapeutic and above drug concentrations as to evaluate the utility of these markers in in vitro systems. Initially, treatment of PHH with any of the drugs caused a statistically significant reduction in enzyme activities to suggest a switch from basic amino acid metabolism towards induced detoxification. However, treatment for prolonged periods of time caused cytolysis, as evidenced by the significant rise in extracellular LDH and the concomitant increase in ALT and AST activity. Notably, amongst the various endpoints studied, urea was best to demonstrate dose dependent metabolic stress, while other markers of hepatocellular injury were highly variable. Taken collectively, urea measurement proofed to be robust in predicting hepatocellular stress; therefore it should be included in preclinical testing strategies for an improved prediction of DILI.