Hydroalcoholic root extracts of Houttuynia cordata (Thunb.) standardized by UPLC-Q-TOF-MS/MS promotes apoptosis in human hepatocarcinoma cell HepG2 via GSK-3ß/ß-catenin/PDL-1 axis.

Houttuynia cordata (Thunb.), an important medicinal plant of Northeast India, Korea, and China, is used to treat various ailments and for anticancer research. Knowing its traditional practices, we are interested in the mode-of-action of HCT on HepG2 to co-relate the traditional practice with modern drug therapeutics. UPLC-Q-ToF-Ms analysis of HCT reveals identification of 14 metabolites. Network pharmacology analysis of the 14 compounds showed interaction with 232 different targets with their potential involvement in hepatocellular carcinoma. Whole extracts impart cytotoxicity on variety of cell lines including HepG2. There was a significant morphological alteration in treated HepG2 cells due to impairment of cytoskeletal components like ß and γ- tubulin. Arrest at G1-S checkpoint was clearly indicated downregulation of Cyclin D1. The root extracts actuated apoptosis in HepG2 as evident from altered mitochondrial membrane potential, Annexin V- FITC, BrdU-PI, AO/EtBr assays, and modulations of apoptotic protein expression but without ROS generation. Whole extracts caused abrogation of epithelial to mesenchymal transition with repression of Snail, N-Cadherin, Vimentin, MMP-9, and upregulation of Pan-Cadherin. Pathway analysis found GSK-3ß in Wnt/ß-Catenin signaling cascade to be involved through Hepatocellular carcinoma (hsa05225) pathway. The GSK-3ß/ß-Catenin/PDL-1 signaling was found to be inhibited with the downregulation of pathway components. This was further confirmed by application of EGF, an inducer of the GSK-3ß/ß-Catenin pathway that neutralized the effect of Houttuynia cordata (Thunb.) root extract on the said pathway. Network pharmacology analysis also confirms the synergy network with botanical-bioactive-target-disease which showed Kaempferol to have the highest degree of association with the said pathway.

Quantification and Standardization of Andrographolide in Andrographis Paniculata Samples by Validated RP-HPLC and HPTLC Methods.

Andrographis paniculata (family Acanthaceae) is known as Kalmegh, one of the traditionally used important medicinal plant contains several biologically active phytochemical including andrographolide. A. paniculata is broadly used by healthcare practitioners in India and also used in different traditional medicinal system. In this study, the leaves of A. paniculata were collected from West Medinipur, East Medinipur, South 24 Parganas, Purulia and Hooghly district of West Bengal, India. This study aiming towards validation and development of a simple, precise and reproducible reverse-phase high-performance liquid chromatography (RP-HPLC) and high-performance thin layer chromatography (HPTLC) methods for quantification of andrographolide in A. paniculata extracts. The validated RP-HPLC and HPTLC study confirmed that different concentrations of andrographolide content present in the plant samples, which are collected from above different districts of West Bengal, India. The amounts of andrographolide were found to be 2.71% (w/w), 3.19% (w/w), 1.83% (w/w), 1.73% (w/w) and 2.94% (w/w) in RP-HPLC study and 2.13% (w/w), 2.51% (w/w), 1.01% (w/w), 1.25% (w/w) and 2.15% (w/w) in HPTLC study. This precise, reproducible, accurate and specific method can be used for the quantification of andrographolide in kalmegh, as per the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines recommendations.

Therapeutic importance of Cucurbitaceae: A medicinally important family.

ETHNOPHARMACOLOGICAL RELEVANCE: Medicinal plants of Cucurbitaceae family consist of several edible fruits and vegetables consumed worldwide since ancient times. The plants of this family have played an essential role in the ethnopharmacological as well as traditional medicinal system globally and their evidence is well established in several traditional literatures. Various plant parts have been used to treat several human ailments viz. Pandu (anemia), Pliharoga (splenomegaly), Sopha (inflammation), Gulma (tumor growth), Adhmana (indigestion. acidity), Garavisa (poisoning) etc. AIM OF THE REVIEW: This review article aims to systematically document and bridge scientific evidences with the ethnopharmacological, ethnoveterinary and folklore claims along with the therapeutic efficacy with mechanism of action found in different literature, books, and scientific articles belonging to the Cucurbitaceae family. MATERIALS AND METHODS: To construct the manuscript a comprehensive literature review was done based on the information collected from Ayurvedic Pharmacopoeia of India; books, research articles and databases such as ScienceDirect, Wiley Online Library, SciFinder, Scopus, Springer, Google Scholar, Web of Science, ACS Publications and PubMed. RESULTS: The plants of Cucurbitaceae family are rich in phytochemicals like terpenoids, glycosides, alkaloids, saponins, tannins, steroids, etc., responsible for the therapeutic effect. Various parts of these plants such as leaves, stems, flowers, fruits, seeds, roots etc. exhibit a plethora of pharmacological activity viz. hypolipidemic, antihyperglycemic, anticancer, antimicrobial, analgesic, anti-inflammatory, anti-stress and immunomodulatory activities. Also, in-vitro and in-vivo reports suggest strong inhibitory potential against α-glucosidase, α-amylase, lipase, carbonic anhydrase enzyme along with antioxidant, anti-inflammatory, antidiabetic, anti-tumor, antifungal, etc. Furthermore many reports suggest these plants are beneficial for nutritional, economical and ethnoveterinary uses. CONCLUSIONS: The current review enlightens the therapeutic potential of the gourd family, comprising of the geographical origins, morphology, phytochemistry, ethnopharmacology, ethnoveterinary, nutritional importance, therapeutic benefits, safety, efficacy and related aspects. The phytochemical and pharmacological potential indicated will popularize this family as a potential source of novel therapeutic agents and functional foods. This study will help to validate the therapeutic claims of several ethnomedicinal uses of this plant family. Furthermore the Cucurbitaceae family needs to be evaluated based on the combine approaches of chemoprofiling and bioexploration to develop the concept of food as medicine for the development of new generation therapeutics leading to the human wellness.

Immunoprotective potential of Ayurvedic herb Kalmegh (Andrographis paniculata) against respiratory viral infections - LC-MS/MS and network pharmacology analysis.

INTRODUCTION: Immunity boosting has emerged as a global strategy to fight the SARS-CoV-2 pandemic situation. In India, AYUSH systems of medicine have been promoted as an immune-protection strategy. Andrographis paniculata (Burm. F) Nees (AP) mentioned in Ayurveda has been widely used for treating sore throat, flu, and upper respiratory tract infections which may provide possible novel therapeutic approaches, exclusively targeting SARS-CoV-2 and its pathways. OBJECTIVES: The present work uses liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolomics and combination synergy analysis based on network pharmacology to mine multimode evidence to understand the possible mechanism of action, diseases association, protein-protein interaction and major pathways involved therein. MATERIAL AND METHODS: Metabolite profiling was performed by Agilent QTOF LC-MS/MS system. Network pharmacology analysis was performed by using functional annotation analysis based on databases like Binding DB, STRING, DAVID and KEGG for further data mining. Further combination synergy was evaluated using "neighbourhood approach" and networks were constructed through Cytoscape 3.2.1. RESULTS: The molecules from kalmegh provides immune-protection and anti-viral response via involving different pathways, like toll-like receptor pathway, PI3/AKT pathway and MAP kinase pathways against COVID-19 infection. The KEGG analysis showed that in a vast majority of the most enriched pathways, AP were associated with viral infections and upper respiratory tract infections. CONCLUSIONS: The results suggest a synergy between andrographolide and other molecules identified as safe and efficacious anti-inflammatory agent having effects on upper respiratory tract infections and can significantly decrease the production of cytokines and pro-inflammatory factors in viral infections.

Evaluation of an ethnomedicinal combination containing Semecarpus kurzii and Hernandia peltata used for the management of inflammation.

CONTEXT: Scientific validation of an ethnomedicinal combination consisting of Semecarpus kurzii Engler (Anacardeaceae) leaves (SKL) and Hernandia peltata Meisn (Hernandeaceae) stem-bark (HPB), traditionally used in ailments related to inflammation, pain and fever. OBJECTIVE: To validate in vivo and in vitro analgesic and antiinflammatory activities of methanol extract of SKL, HPB and their combination. MATERIALS AND METHODS: Analgesic activity was tested by acetic acid induced writhing reflex and tail flick in Swiss albino mice, while the anti-inflammatory activity was studied in acute, subacute and chronic model on Wistar rats. The vascular permeability, membrane stabilization and protein denaturation were examined to know the possible mode of action. RESULTS: Significant (p < 0.01) analgesic (78.04% inhibition of writhing) and antiinflammatory (72.54% inhibition of paw edema) activity was observed in combination of SKL and HPB extracts at 250 mg/kg each. The SKL extract alone inhibits acetic acid-induced vascular permeability (64.4%) at 500 mg/kg, while in combination at 250 mg/kg each, the inhibition was 69.49% (p < 0.01). Furthermore, SKL in combination with HPB (0.25 mg/mL each) prevent RBC hemolysis (61.91%) and inhibition of protein denaturation (76.52%)-like indomethacin. DISCUSSION AND CONCLUSION: The SKL and HPB extract, alone (500 mg/kg) and in combination, (250 mg/kg each) had significant analgesic and antiinflammatory activity, probably by inhibiting the release of certain inflammatory mediators and membrane stabilization, due to the presence of triterpenes, tannins and related phytochemicals in the extracts. Thus, our results demonstrated that this combination provide the scientific rationale of its folk use.

Antitumor activity and antioxidant role of Streblus asper bark against Ehrlich ascites carcinoma in Swiss albino mice.

The present study assessed the methanol extract of Streblus asper stem bark (MESA) for antitumor effect and antioxidant influence against Ehrlich ascites carcinoma (EAC) in Swiss albino mice. Twenty four hours after intraperitonial inoculation of tumor (EAC) cells in mice, MESA was administered at 200 and 400 mg/kg body weight daily for 9 consecutive days. On the 10 th day, half of the mice were sacrificed for estimation of tumor parameters, haematological, liver and kidney antioxidant parameters; and the rest were kept alive for assessment of survival. MESA exhibited dose dependent and significant (p < 0.01) decrease in tumor proliferation and extended the life span of EAC bearing mice. Hematological profiles were significantly (p < 0.01) restored near to normal in MESA treated mice as compared to EAC control. MESA treatment significantly (p < 0.01) modulated the hepatic and renal antioxidant parameters as compared to EAC control. The present study demonstrated that S. asper bark possessed promising antitumor efficacy in mice, plausibly mediated by amelioration of oxidative stress by multiple mechanisms.

Antitumour activity of Terminalia arjuna leaf against Ehrlich ascites carcinoma in mice.

Terminalia arjuna Roxb. (Combretaceae), commonly known as 'Arjuna', is a large tree occurring throughout the Indian peninsula. This study was undertaken to evaluate the methanol extract of T. arjuna leaf (META) for antitumour activity against Ehrlich ascites carcinoma (EAC) in Swiss albino mice. Twenty-four hours after intraperitonial inoculation of tumour (EAC) cells in mice, META was administered at 100 and 200 mg kg(-1) body weights for 9 consecutive days. On day 10, half of the mice were sacrificed and the rest kept alive for an assessment of the increase in life span. The antitumour effect of META was assessed by evaluating tumour volume, tumour weight, viable and non-viable tumour cell counts, median survival time and increase in life span of EAC-bearing hosts. Haematological profiles were estimated. META showed a significant (p<0.001) decrease in tumour volume, tumour weight and viable cell count, and also increased the life span of EAC-bearing mice. Haematological profiles were significantly (p<0.001) restored to normal levels in META-treated mice compared to the EAC control. Therefore, from this study, it can be concluded that T. arjuna leaf exhibited remarkable antitumour activity against EAC in Swiss mice.

Antitumor efficacy and amelioration of oxidative stress by Trichosanthes dioica root against Ehrlich ascites carcinoma in mice.

CONTEXT: Trichosanthes dioica Roxb. (Cucurbitaceae) is a dioecious climber, traditionally used in India for several medicinal purposes. OBJECTIVE: The present study assessed the hydroalcoholic extract of T. dioica root (TDA) for antitumor effect and antioxidant influence against Ehrlich ascites carcinoma (EAC) in Swiss albino mice. METHODS: Twenty four hours after intraperitoneal inoculation of tumor (EAC) cells in mice, TDA was administered at 5 and 10 mg/kg body weight daily for 9 consecutive days. On the 10th day, half of the mice were sacrificed for estimation of tumor proliferation, hematological, and liver antioxidant parameters viz. lipid peroxidation, reduced glutathione (GSH), glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT); and the rest were kept alive for assessment of increase in life span. The antitumor effect of TDA was assessed by evaluating tumor weight, tumor volume, packed cell volume, viable and non-viable tumor cell counts, median survival time and percentage increase in life span of EAC bearing mice. RESULTS AND DISCUSSION: TDA exhibited dose dependent and significant (p < 0.001) decrease in tumor weight, tumor volume, packed cell volume and viable cell count and extended the life span of EAC bearing hosts. Hematological profiles were significantly (p < 0.001) restored near to normal in TDA treated mice as compared to EAC control. TDA treatment significantly (p < 0.001) modulated the aforesaid liver antioxidant parameters as compared to EAC control. CONCLUSION: The present study demonstrated that TDA possessed promising antitumor efficacy in mice, plausibly mediated by amelioration of oxidative stress by multiple mechanisms.

Hypoglycemic activity of Erythrina variegata leaf in streptozotocin-induced diabetic rats.

CONTEXT: Erythrina variegata Linn. (Fabaceae), commonly known as Tiger's Claw, is a thorny deciduous tree grown in tropical and subtropical regions of Eastern Africa, Southern Asia, and Northern Australia. In India, its leaves are traditionally used for diabetes mellitus. OBJECTIVE: To evaluate the hypoglycemic activity of methanol extract of E. variegata leaf (MEEV) in streptozotocin (STZ)-induced diabetic Wistar rats. MATERIALS AND METHODS: Hyperglycemia was induced in rats by single intraperitoneal injection of STZ (55 mg/kg body weight). Three days after STZ induction, the hyperglycemic rats were treated with MEEV orally at the doses of 300, 600, and 900 mg/kg body weight daily for 21 days. Glibenclamide (1 mg/kg, orally) was used as reference drug. The fasting blood glucose levels were measured on every 7th day during the 21 days of treatment. Serum biochemical parameters including lipid content were estimated. RESULTS AND DISCUSSION: MEEV at the doses of 300, 600, and 900 mg/kg orally significantly (P < 0.01) and dose-dependently reduced and normalized blood glucose levels as compared to that of STZ control group; the dose 900 mg/kg being the most potent showing complete normalization of blood glucose levels. Serum biochemical parameters including lipid profile were significantly (P < 0.01) restored toward normal levels in META-treated rats as compared to STZ control animals. CONCLUSION: This study concludes that E. variegata leaf demonstrated promising hypoglycemic action in STZ-induced diabetic rats substantiating its ethnomedicinal use.

Hepatoprotective activity of Cyperus tegetum rhizome against paracetamol-induced liver damage in rats.

In present study the methanol extract of Cyperus tegetum rhizome (MECT) was evaluated for its effect on paracetamol-induced liver damage in Wistar rats. Serum biochemical parameters viz. serum glutamine oxaloacetate transaminase (SGOT), serum glutamine pyruvate transaminase (SGPT), serum alkaline phosphatase (ALP), total serum protein, total bilirubin content and liver biochemical parameters such as thiobarbituric acid reactive substances (TBARS) and reduced glutathione content were estimated. Biochemical and histopathological observations indicated that MECT had remarkable hepatoprotective effect against paracetamol-induced liver damage in rats.

Evaluation of analgesic and anti-inflammatory activity of diospyros cordifolia extract.

In this study we evaluated the analgesic and anti-inflammatory activities of the methanol extract of stem bark of Diospyros cordifolia (MEDC) Roxb. The analgesic effects of the stem bark of the plant was assessed in mice using the tail-flick method while carrageenan, histamine and dextran induced paw oedema was used to study the antiinflammatory effects in rats. The MEDC exhibited significant (p<0.01) analgesic effects comparable to the reference drug diclofenac sodium. MEDC also was evaluated for its anti-inflammatory potential against carrageenan, histamine and dextran induced rat paw edema. The methanol extract (25 and 50 mg / kg body weight) exhibited significant (p<0.01) activity against all phlogistic agents used in a dose dependent manner. All these effects were compared with reference drug phenylbutazone (100 mg/kg body weight).

Evaluation of analgesic and anti-inflammatory activity of Diospyros Cordifolia Extract

Abstract: In this study we evaluated the analgesic and anti- inflammatory activities of the methanol extract of stem bark of Diospyros cordifolia (MEDC) Roxb. The analgesic effects of the stem bark of the plant was assessed in mice using the tail-flick method while carrageenan, histamine and dextran induced paw oedema was used to study the antiinflammatory effects in rats. The MEDC exhibited significant (p<0.01) analgesic effects comparable to the reference drug diclofenac sodium. MEDC also was evaluated for its anti-inflammatory potential against carrageenan, histamine and dextran induced rat paw edema. The methanol extract (25 and 50 mg / kg body weight) exhibited significant (p<0.01) activity against all phlogistic agents used in a dose dependent manner. All these effects were compared with reference drug phenylbutazone (100 mg/kg body weight)

Evaluation of anticancer activity of Cleome gynandra on Ehrlich's Ascites Carcinoma treated mice.

ETHNOPHARMACOLOGICAL RELEVANCE: The plant Cleome gynandra L. (Capparidaceae), is commonly known as 'Hurhur'and 'Karaila' in India and 'Cat's whiskers' in English. Traditionally the whole plant is used in the treatment of tumor, anti-inflammatory and lysosomal stability actions. AIM OF STUDY: The objective of present study is to explore the anticancer activity of the methanol extract of the Cleome gynandra in Swiss albino mice against Ehrlich Ascites Carcinoma (EAC) cell line. MATERIALS AND METHODS: Anticancer activity of methanol extract of Cleome gynandra (MECG) was evaluated in Swiss albino mice against Ehrlich Ascites Carcinoma (EAC) cell line at the doses of 200 and 400mg/kg body weight intraperitoneally. MECG was administered for nine consecutive days. Twenty-four hours of last dose and 18 h of fasting, the mice were sacrificed and antitumor effect of MECG assessed by evaluating tumor volume, viable and nonviable tumor cell count, tumor weight and hematological parameters of EAC bearing host. RESULTS: MECG showed significant decrease in (p<0.01) tumor volume, viable cell count, tumor weight and elevated the life span of EAC tumor bearing mice. Hematological profile such as RBC, hemoglobin, WBC and lymphocyte count reverted to normal level in MECG treated mice. CONCLUSION: From the result it was showed that the extract has potent dose dependent anticancer activity and that is comparable to that of 5-fluorouracil.