Comparative analysis of the seasonal influence on polyphenolic content, antioxidant capacity, identification of bioactive constituents and hepatoprotective biomarkers by in silico docking analysis in Premna integrifolia L.

The present study reports the effect of different seasons on polyphenol content and antioxidant potential of ethanolic, methanolic, ethyl acetate, and aqueous extracts of leaves, stems, and roots of Premna integrifolia. Ethyl acetate extract of leaves (EAEPI) collected in the rainy season showed potent antioxidant activity with highest total phenol (74.33 ± 2.26 µg/mg, gallic acid equivalent), and flavonoid (98.83 ± 0.26 µg/mg, rutin equivalent) content. Therefore, EAEPI extract was subjected to characterization by UHPLC-Q-TOF-MS/MS and GC-MS analysis for the identification of active constituents. UHPLC-Q-TOF-MS/MS analysis in + ve ion mode revealed the presence of eight polyphenolic compounds namely quercetin-3-D-xyloside, kaempferol-3,7-O-bis-alpha-L-rhamnoside, isorhamnetin-3-Oglucoside, luteolin-3',7-di-O-glucoside, eriodictyol-7-O-glucoside, syringetin-3-O-galactoside, petunidin-3-O-beta-glucopyranoside and vitexin-2″-O-rhamnoside. GC-MS analysis confirmed the presence of 26 compounds with six major compounds viz; citronellol, phytol acetate, campesterol, squalene, stigmasterol, and hexadecanoic acid. These compounds are reported for the first time from P. integrifolia except phytol and stigmasterol. Our previous study validates the hepatoprotective potential of P. integrifolia but there was no idea about the bioactive compound responsible for the activity. So, in present work, the major compounds identified in spectrometry analysis were subjected to in silico docking against an important liver enzyme alanine amino transaminase to confirm its hepatoprotective properties. Docking analysis validates the presence of two hepatoprotective lead compounds stigmasterol, and campesterol, which satisfy the drug-likeness criteria with good absorption, distribution, metabolism, and toxicity properties. Thus, present work gives a clear insight about the influence of season on the total polyphenolic constituent in different plant parts of P. integrifolia, their antioxidant potential and preclinical evaluation of hepatoprotective lead compounds.

Phytochemical profiling and cytotoxic evaluation of Premna serratifolia L. against human liver cancer cell line.

Premna serratifolia L. (Lamiaceae) is a medicinal plant, widely distributed in the tropical and subtropical regions and commonly used in traditional medicine. The current study was focused to evaluate the cytotoxic potential of aqueous extract of root of P. serratifolia (AEPS) against human hepatoblastoma cancer cell line (Hep G2).The yield of the dried extract was 5.8% and used for further studies.Cytotoxic potential of AEPS was analyzed by MTT assay, which exhibits IC50 value 1000 µg/mL after 48 h incubation. Hoechst and AO/EtBr staining, ROS measurement, mitochondrial membrane potential, clonogenic and wound healing assays also confirmed the cytotoxic efficacy of AEPS in dose and time-dependent manner. UPLC-Q-TOF-MS/MS analysis of AEPS confirmed the presence of 12polyphenolic compounds, namely 4-hydroxy-3-methoxycinnamic acid, linarin, peonidin-3,5-O-di-beta-glucopyranoside, diosmin, trans-cinnamic acid, daidzein, saponarin, homoorietin, acacetin, sarsasapogenin, phytol and sissotrin. The cytotoxic potential of AEPS might due to presence of biologically active polyphenolic compounds.

Toxicity profiling and antioxidant activity of ethyl acetate extract of leaves of Premna integrifolia L. for its application as protective agent against xenobiotics.

Premna integrifoliaL. (Lamiaceae) is widely used in herbal formulation "Dashmoolarishta" which is useful in postnatal care. Ethyl acetate extract obtained from the leaves was evaluated for phenolic content and its antioxidant activity. Acute and subacute toxicity of the extract was studied in mice of both sexes to get an idea about LD50 value and assessed its safety profile before its application as a protective agent against different toxicities induced by xenobiotics. Phenol enriched extract (phenol content is 63.10 ± 1.26 mg/g of gallic acid equivalent and flavonoid content 75.33 ± 0.23 mg/g of rutin equivalent) showed good antioxidant activity. In acute toxicity studies it was observed that single different doses (300-5000 mg/kg b.wt.) of extract did not show any mortality of mice. Thus the LD50 of the extract was determined, and it was higher than 5000 mg/kg. There was no major change in behavioral and general appearance of mice. External morphology of liver, kidneys, lungs, spleen and heart did not show any effect of treatment. In subacute toxicity no statistically significant change in body weight, relative organ weight, food intake and water uptake, hematological, biochemical parameters were reported after comparison with control. Extract did not show significant effect in the level of antioxidant enzymes in the liver of mice of treated groups. No histopathological changes were observed in liver and kidney tissues. Thus, extract did not show any sign of toxic effects, when administered orally to male and female mice at dose level up to 1000 mg/kg. So, it can be utilized as protective agent against toxicity produced by different xenobiotics.

Premna integrifolia ameliorates cyclophosphamide-induced hepatotoxicity by modulation of oxidative stress and apoptosis.

The present study was designed to evaluate the ameliorative effect of ethyl acetate extract of Premna integrifolia L. (EAEPI) leaves in cyclophosphamide (CP)-induced hepatic injury in mice. Mice were intoxicated with CP (200 mg/kg b. wt., i.p.) for 5 weeks or EAEPI (400 and 600 mg/kg b. wt., orally) in combination with CP. The results demonstrated that EAEPI exerts protective effect against CP induced hepatotoxicity, as evident from restoration of altered haematological parameters and alleviations of liver marker enzymes in serum. EAEPI also attenuated oxidative stress and antioxidant markers as evident from reversal of lipid peroxidation, glutathione levels as well as activities of catalase and superoxide dismutase enzymes. Moreover, EAEPI attenuated apoptosis and histopathological liver tissue damage in CP-intoxicated mice. In conclusion, EAEPI could protect mice liver against cyclophosphamide toxicity by inhibiting oxidative stress and apoptosis.The protective activity of EAEPI may be due to presence of polyphenolic compounds as identified by UHPLC-Q-TOF-MS/MS.