ß-sitosterol isolated from the leaves of Trema orientalis (Cannabaceae) promotes viability and proliferation of BF-2 cells.

Trema orientalis is a pioneer species in the cannabis family (Cannabaceae) that is widely distributed in Thai community forests and forest edges. The mature leaves are predominantly used as an anti-parasite treatment and feed for local freshwater fish, inspiring investigation of their phytochemical composition and bioactivity. The purpose of this work was to investigate the bioactive compounds in T. orientalis leaf extract and their cytotoxicity in the BF-2 fish cell line (ATCC CCL-91). Flash column chromatography was used to produce 25 mL fractions with a mixture solvent system comprised of hexane, diethyl ether, methanol, and acetone. All fractions were profiled with HPLC-DAD (mobile phase methanol:aqueous buffer, 60:40 v/v) and UV detection (wavelengths 256 and 365 nm). After drying, a yellowish powder was isolated from lipophilic leaf extract with a yield of 280 µg/g dry weight. Structure elucidation by nuclear magnetic resonance (NMR) indicated it to consist of pure ß-sitosterol. The lipophilic extract and pure compound were evaluated for cytotoxicity using BF-2 cells. MTT assays showed both leaf extract and pure compound at 1 µg/mL to increase cell viability after 24 h treatment. The respective half maximal inhibitory concentration (IC50) values of leaf extract and ß-sitosterol were 7,027.13 and 86.42 µg/ml, indicating a lack of toxicity in the BF-2 cell line. Hence, T. orientalis can serve as a source of non-toxic natural lipophilic compounds that can be useful as bioactive ingredients in supplement feed development.

Trema orientalis (Linn.) leaves promotes anticancer activity in Ehrlich ascites carcinoma (EAC) in Swiss albino mice.

Background The in vivo anticancer effect of the Trema orientalis leaves crude methanol extract (TLME) was screened against Ehrlich ascites carcinoma (EAC) in Swiss albino mice. Materials and methods The cytotoxic activity of TLME was determined in vitro by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The growth inhibitory activity and morphological alterations were determined by the hemocytometer counting of the EAC cells using trypan blue dye. The apoptotic cells were assessed by DAPI (4',6-diamidino-2-phenylindole) staining. The hematological and biochemical parameters of experimental mice were also estimated. Results After treatment with the TLME, the viable tumor cell count, morphological changes and nuclear damages of the EAC cells were observed along with the hematological parameters of the experimental mice. The LD50 of TLME was 3120.650 mg/kg body weight, and this extract was proven to be safe at a dose of as high as 800 mg/kg body weight. The oral administration of the TLME at 400 mg/kg body weight resulted in approximately 59% tumor cell growth inhibition compared with the control mice, with considerable apoptotic features, including membrane blebbing, chromatin condensation, nuclear fragmentation and aggregation of the apoptotic bodies in DAPI staining under a fluorescence microscope. The TLME also dose-dependently restored the altered hematological parameters to approximately normal levels. The TLME exhibited bolstering cytotoxic effect against the EAC cell with the IC50 value of 29.952 ± 1.816 µg/mL. Conclusion The TLME has potential as a natural anti-cancer product with apoptosis induction property and cytotoxicity against carcinoma cells.

Mosquito larvicidal potential of four common medicinal plants of India.

BACKGROUND & OBJECTIVES: Mosquitoes transmit serious human health diseases, causing millions of deaths every year. Plants may be sources of alternative mosquito control agents. The present study was carried out to assess the role of larvicidal activities of the crude extracts of four plants viz. Alternanthera sessilis L. (Amaranthaceae), Trema orientalis L. (Cannabaceae), Gardenia carinata Smith. (Rubiaceae) and Ruellia tuberosa L. (Acanthaceae) against Culex quinquefasciatus Say in laboratory bioassay. METHODS: Selective concentrations (0.5, 1 and 1.5%) of crude extract of all four plant leaves were tested against I st to IV th instar larvae of Cx. quinquefasciatus. Log probit analysis (at 95% confidence level) revealed the LC50 values. Preliminary qualitative phytochemical analyses of crude extracts were also done. The lethal concentrations (%) of crude extracts at 24 h against III rd instar larvae were also studied on non-target organisms. RESULT: In a 72 h bioassay experiment with crude extract, the highest mortality was recorded in 1.5 per cent extract. A. sessilis showed the highest mortality (76.7%) at 1.5 per cent crude extract against II nd instar larvae having LC50 value of 0.35 per cent, followed by R. tuberosa (LC50 = 1.84%), G. carinata (LC50 = 2.11) and T. orientalis (LC50 = 2.95%). The regression equation showed a dose-dependent mortality, as the rate of mortality (Y) was positively correlated with the concentration (X). Phytochemical analysis of the crude extract showed the presence of many bioactive phytochemicals such as steroids, alkaloids, terpenes, saponins, etc. No changes in the swimming behaviour and survivality of non-target organism were noticed at the studied concentrations. INTERPRETATION & CONCLUSIONS: Crude extract of the four selected plants showed larvicidal activity against Cx. quinquefasciatus. The extracts at the studied concentrations did not produce any harmful effect on non-target organisms.

Trema micrantha toxicity in horses in Brazil.

After ingesting green leaves of T. micrantha, 2 horses showed apathy, locomotor deficit, blindness, recumbency, paddling, coma and death. The main gross findings were scattered haemorrhages, enhanced lobular pattern of the liver, and cerebral oedema. Histological changes included disseminated haemorrhages, massive hepatocellular necrosis, neuronal degeneration, Alzheimer type II astrocytes and cerebral perivascular oedema. Clinicopathological findings which were comparable with those observed in Trema micrantha poisoned ruminants, associated with epidemiological evidence suggested the diagnosis.Trema micrantha poisoning should be evaluated as a possible cause in the diagnosis of equine hepatopathy and occasional secondary encephalopathy.

Glucose lowering efficacy of the aqueous stem bark extract of Trema orientalis (Linn) Blume in normal and streptozotocin diabetic rats.

The glucose-lowering efficacy of the aqueous stem bark extract of Trema orientalis (Ulmaceae) was evaluated both in normal and streptozotocin-induced diabetic rats. In normoglycemic rats, the single oral administration of the aqueous extract of T. orientalis failed to reduce blood glucose levels while in STZ-diabetic rats, the plant extract (38-300 mg/kg) exhibited significant hypoglycaemic activity with a maximum effect of 29.67%, 5 hours after administration of the 75 mg/kg dose when compared with the diabetic untreated group. Glibenclamide was not able to lower blood glucose in STZ-diabetic rats, while it significantly lowered the blood sugar in normoglycemic rats. The hypoglycaemic property of T. orientalis was also assessed by an oral glucose tolerance test (OGTT) in STZ-diabetic rats. The aqueous extract of T. orientalis and the reference drug, glibenclamide, (10 mg/kg) produced significant blood glucose lowering effects in the diabetic rats when compared to the diabetic controls. One week after repeated administration of T. orientalis extract, blood glucose levels were significantly decreased (p < 0.05) and still remained low after 2 weeks (p < 0.01). The results indicated that T. orientalis stem bark extract significantly reduces blood glucose in STZ-induced diabetic rats by a mechanism different from that of sulfonylurea agents. The present investigation provides pharmacological evidence that the use of this plant extract in traditional medicine for cardiovascular disease can be of benefit particulary in diabetic patients.

[Studies on chemical constituents from branch of Trema angustifolia].

OBJECTIVE: To study the chemical constituents from the branch of Trema angustifolia. METHOD: The compounds were isolated by silica gel column chromatography, and their structures were elucidated by means of spectral analysis. RESULT: Six compounds were identified as beta-sitosterol (I), viminalol (II), acetylursolic acid (III), oleanolic acid (IV), paprazine (V), moupinamide (VI). CONCLUSION: All the compounds were isolated from this plant for the first time.