@article {598, title = {Purified Anthocyanin from in vitro Culture of Bridelia retusa (L.) Spreng. Capable of Inhibiting the Growth of Human Oral Squamous Cell Carcinoma Cells}, journal = {Pharmacognosy Journal}, volume = {10}, year = {2018}, month = {March 2018}, pages = {559-566}, type = {Original Article}, chapter = {559}, abstract = {

The present study aims in vitro cell suspension culture of Bridelia retusa, isolation of anthocyanin, purification, fractionation and its anti-metastatic potential against oral squamous carcinoma cells. Experimental results reveal that 2, 4-D either alone or in combination with kinetin supplemented in MS medium showed significant initiation of callus from leaf explants than stem. Growth hormones, pH, light, and carbon source influence anthocyanin synthesis. Maximum callus induction was noticed with 2.5 mg/L N6-benzyladenine (BA) + 2 mg/L 2, 4-dichlorophenoxyacetic acid (2, 4-D) (98.9\%). Fresh and dry weight of the calli were i.e., 1.9 \± 0.04 and 0.45 \± 0. 03 g respectively. Optimal response was seen with light on MS medium contain 4\% glucose + 2.5 mg/L BA and 2 mg/L 2, 4-D at pH 3.5 yielded 2.8 mg /g of anthocyanins. Suspension culture medium fortified with 2, 4-D (2.5 mg/L) + BA (2 mg/L) at pH 5.0 induced anthocyanin production at pH 4.4 \– 4.6. HCl-ethanol extraction for 90 min yielded the maximum anthocyanin content. Fractionation of anthocyanin using HPLC coupled with mass spectrometry revealed 07 fractions such as acylated cyanidins, two peonidins, cyanidin 3-p-coumaroyl and feruloyl diglucoside-5-glucosides. In the search of novel therapeutic drugs against cancer, cytotoxicity effect of B.retusa anthocyanin extracts on human oral squamous cell carcinoma (SCC4, SCC9 and SCC25) cells using cell adhesion and cell viability assay was carried. The morphological alterations in SCCs cells after treatment with B.retusa anthocyanin includes nuclear condensation, fragmentation and apoptotic cells as revealed by Hoechst stain. Flow cytometry showed arresting of SCC25 cells mostly in the G0/G1 and S-G2/M stages with a concomitant up regulation of sub-G1 fraction, indicating cell death by apoptosis. Apoptosis was further substantiated by the activation of caspase-3 expression in the SCC25 cells treated with B.retusa anthocyanin. Thus, it is possible to suggest that B.retusa anthocyanin cause apoptosis of SCCs and warrant further investigation using animal models.

}, keywords = {Anthocyanin, Anti-metastatic potential, Apoptosis, Bridelia retusa, Cell suspension, in vitro culture, Purification}, doi = {10.5530/pj.2018.3.91}, url = {http://fulltxt.org/article/524}, author = {Aswathy Jayasree Madanakumar and Bosco Lawarence and Manoj GS and Murugan Kumaraswamy} } @article {378, title = {Comprehensive Evaluation of Antioxidant Potential of Selected Osbeckia species and their in vitro Culture, Purification and Fractionation}, journal = {Pharmacognosy Journal}, volume = {9}, year = {2017}, month = {July 2017}, pages = {674-682}, type = {Original Article}, chapter = {674}, abstract = {

Background: Health-benefit properties of natural pigments have been intensely studied, especially the anthocyanins. In the last few decades, research on anthocyanins has attracted biologists by the increasing evidence of their health beneficial effects. Osbeckia, belongs to Melastomataceae and is well-known for colouring pigments and other bioactive compounds. In the present study, total anthocyanin and antioxidant capacity indicators were evaluated from 8 Osbeckia spp. and anthocyanin was extracted from in vitro cultures of O. aspera and O. reticulata. Materials and Methods: The antioxidant effect was studied using ABTS (2, 2\’-azino-bis-3-ethyl benzthiazoline-6-sulphonic acid) radical cation decolourisation assay, the FRAP, the scavenging ability of hydroxyl radicals and the superoxide anion scavenging activity. Anthocyanin extracted from in vitro cultures were purified and fractionated using column chromatography and LC-MS MS analysis. Results: In vitro cultures of O. aspera was obtained in MS medium fortified with various combinations of Benzyl Adenine (BA), Naphthalene acetic acid (NAA) and 2, 4-D. The chromatograms of O. aspera revealed the presence of malvidin-3 -diglucoside, peonidin, delphinidin and cyanindin whereas O. reticulata cultures accumulated large amounts of malvidin, cyanindin and cyanidin aglycone. The purified anthocyanins of these species were evaluated for their antioxidant potential and was found more remarkable than the crude extracts. Conclusion: Osbeckia species are rich in anthocyanin and therefore display potential AOX power. O. aspera and O. reticulata callus was induced in vitro production of anthocyanins. The pool of anthocyanins was purified and fractionated by LCMS/ MS and AOX assays were performed with the purified anthocyanin which showed higher level activities.

}, keywords = {Anthocyanins, Antioxidant Capacity, Free Radicals, Osbeckia Spp., Reactive Oxygen Species}, doi = {10.5530/pj.2017.5.107}, url = {/files/pj-9-5/10.5530pj.2017.5.107/index.html}, author = {Bosco Lawarence and Murugan K} } @article {443, title = {Isolation, Purification of Quercetin from in vitro Cell Suspension Culture of Caesalpinia pulcherrima and its Analysis by HPLC-DAD and NMR}, journal = {Pharmacognosy Journal}, volume = {9}, year = {2017}, month = {November 2017}, pages = {s44-s51}, type = {Original Article}, chapter = {s44}, abstract = {

Background: Caesalpinia pulcherrima, belongs to Caesapiniaceae, is a known medicinal plant widely distributed in India and is used in traditional medicine for the treatment of various ailments. Many phytochemicals are reported from the plant as potential source of crude drug. Materials and Methods: An efficient and simple reproducible protocol was developed for callus production using leaf explants of C. pulcherrima. The combination of 2, 4-D, kin and BA, was used for the callus induction. Subsequently, cell suspension culture and quercetin synthesis from in vitro callus was attempted. Role of effect of elicitors (Sucrose, ABA and salicylic acid) in cell suspension culture was carried in MS medium containing 2,4-D + BA + kinetin. Flavonoids was purified, fractionated by HPLC-DAD and NMR. Results: 2, 4-D (2.5 mg/L), BA (2.5 mg/L) + kin (1 mg/mL) was effective for maximum callus induction from leaf explants. Significant cell suspension culture was noticed with liquid MS medium containing 2,4-D (2 mg/L)+ BA (1mg/L)+ kinetin (1.5 mg/L). Sucrose, ABA and salicylic acid (SA) at different concentrations influenced cell biomass and quercetin accumulation. The addition of ABA/SA along with sucrose was found to have no remarkable effect on cell biomass and also quercetin synthesis. However, cells cultured in the medium fortified with 45 g/L sucrose without ABA/ SA showed the highest quercetin content (16.5 mg/g). Flavonoids was purified, fractionated by HPLC-DAD and NMR revealed the presence of 9 components such as quercetin, isoquercetin, quercetrin, rutin, quercetin 3-O-\β-D-xyloside, quercetin 3-Oarabinopyranoside, quercetin 3-O- \α-arabinopyranosyl (1\→2) \β-galactopyranoside, isorhamnetin 3-O-rutinoside and an unknown compound. Conclusion: C. pulcherima reveals significant synthesis of quercetin. Quercetin content recorded in cell suspension culture was significantly higher compared with in vivo plants grown in fields and the compounds were identified by NMR.

}, keywords = {Caesalpinia pulcherrima, Callus, Cell suspension culture, Elicitors; growth hormones, Quercetin}, doi = {10.5530/pj.2017.6s.156}, url = {http://fulltxt.org/article/380}, author = {Aswathy Jayasree Madanakumar and Greeshma Murukan and Bosco Lawarence and Murugan Kumaraswamy} }