Immunomodulatory effects of proanthocyanidin A-1 derived in vitro from Rhododendron spiciferum

The objective of this study was to evaluate the immunomodulatory effects of proanthocyanidin A-1 (PAA-1) from leaves of Rhododendron spiciferum (Ericaceae). In vitro tests showed that PAA-1 stimulated cell proliferation of splenocytes and peritoneal macrophages significantly enhanced the cytotoxicity of natural killer (NK) cells and increased CD4⁺ and CD8⁺ cell populations. PAA-1 also regulated the expression of Th1- and Th2-related cytokines. Moreover, this study showed that PAA-1 exhibited a significant effect on NBT dye reduction and lysosomal enzyme activity responses in macrophages, indicating effective phagocytic activation. These results revealed that PAA-1 exhibits immunomodulatory activity without a clear dose response.     

Echinacoside promotes bone regeneration by increasing OPG/RANKL ratio in MC3T3-E1 cells

Echinacoside (ECH), isolated from Cistanche tubulosa (Schrenk) R. Wight stems, was subjected to in vitro experiments to investigate its bioactivities on proliferation, differentiation and mineralization of the osteoblastic cell line MC3T3-E1. MTT assay, the alkaline phosphatase (ALP) activity and calcium deposition were determined, and the secretion of collagen I (COL I), osteocalcin (OCN), osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were also assayed by enzyme-linked immunosorbent assay (ELISA). The results showed that ECH caused a significant increase in cell proliferation, ALP activity, COL I contents, OCN levels and an enhancement of mineralization in osteoblasts at the concentration range from 0.01 to 10 nmol·L^− 1 (p < 0.05), suggesting that ECH has a stimulatory effect on osteoblastic bone formation or has potential activity against osteoporosis. In addition, the ratio of OPG/RANKL also could be enhanced by ECH. These findings provide the potent evidence that ECH can promote bone regeneration in cultured osteoblastic MC3T3-E1 cells, which might be done by elevating the OPG/RANKL ratio, and potential evidence for echinacoside to be a promising drug or a lead compound in the development of disease-modifying drug to prevent osteoporosis.     

Three new sesquiterpenes from Tithonia diversifolia and their anti-hyperglycemic activity

Three new germacrane sesquiterpenes (1), (2), (3), along with eleven known sesquiterpenes, namely, tirotundin-3-O-methyl ether (4), deacetylvguiestin (5), 1β-hydroxydiversifolin-3-O-methyl ether (6), tagitinin C (7), 1β-hydroxytirotundin-3-O-methyl ether (8), 1β-hydroxytirotundin-1,3-O-dimethyl ether (9), tagitinin F-3-O-methyl ether (10), tagitinin F (11), tagitinin A (12), 3β-acetoxy-4α-hydroxyeduesm-11(13)-en-12-oic acid (13) and ilicic acid (14) were isolated from the aerial parts of Tithonia diversifolia. Their structures were established by spectroscopic analysis, while the relative configuration of compound 1 was confirmed by X-ray diffraction analysis. In addition, compounds 1–14 were evaluated in vitro for their anti-hyperglycemic activity by glucose uptake in 3T3-L1 adipocytes. It was found that 10 μg/mL 1, 3, 6 and 8 could significantly increase glucose uptake without significant toxic effects.     

N-Methyl-6, 7-dimethoxyisoquinolone in Annona squamosa twigs is the major immune modifier to elicit polarized Th1 immune response in BALB/c mice

Annona squamosa (AS) has traditionally been used as ethnomedicine. We have earlier extracted and fractionated the twigs of AS based upon its bioactivity and observed its immune potentiating activity that was localized in its three fractions. Present communication deals with the phytochemical analysis and pharmacological investigation of the most active chloroform fraction that led to isolation and identification of a number of compounds whose structures were elucidated using 1D and 2D NMR spectroscopic analysis. Amongst the twelve pure compounds isolated, five compounds Lanuginosine (1), (+) -O- methylarmepavine (2), (+)-anomuricine (3), Isocorydine (4), and N-methyl-6, 7-dimethoxyisoquinolone (5) were evaluated in vivo for their immune modifier activities in BALB/c mice after oral administration at three log doses of 0.3, 1.0 and 3.0 mg/kg for 14 consecutive days. Of these, three compounds (1, 2 and 5) showed dose dependent immune stimulating activity. However, the uppermost activity was noted in the compound N-methyl-6, 7-dimethoxyisoquinolone at the 3.0 mg/kg oral dose. The activity was assessed in the form of increased splenic T and B cellular proliferation, up-regulated CD4+, CD8+ and CD19+ cell population and accentuation in the peritoneal macrophage function. The compound possibly acted modifying the expression of Th1- and Th2- cytokines via stimulation of pro-inflammatory Th1 cytokines IL-2 and IFN-γ. These results warrant the use of the above compounds as an efficient immune-stimulant or immune-adjuvant against diseases with immune suppression. The analogs of the compound may further be chemically synthesized to achieve desired immune modifying activity.     

New amino butenolides from the bulbs of Fritillaria unibracteata

Five new amino γ-butenolides, fritenolide A (1), B (2), C (3), D (4), and E (5), along with four known compounds, were isolated from the bulbs of Fritillaria unibracteata. Their structures were determined by spectroscopic analysis, including 1D NMR, 2D NMR, HRESIMS, HRESIMS/MS, IR, and CD techniques. All isolates were evaluated for the protective activity on injured hepatocytes and cytotoxic activity on human cancer cells in vitro. The unusual amino butenolides were isolated from the Liliaceae family for the first time.     

Protective effect of Codium fragile against UVB-induced pro-inflammatory and oxidative damages in HaCaT cells and BALB/c mice

Acute exposure to ultraviolet (UV) radiation causes pro-inflammatory responses via diverse mechanisms including oxidative stress. Codium fragile is a green alga of Codiales family and has been reported to exhibit anti-edema, anti-allergic, anti-protozoal and anti-mycobacterial activities. In this study, we have investigated a novel anti-inflammatory potential of C. fragile using in vitro cell culture as well as in vivo animal models. In HaCaT cells, buthanol and ethylacetate fractions of 80% methanol C. fragile extract (CFB or CFE) and a single compound, clerosterol (CLS) isolated from CFE attenuated UVB (60 mJ/cm²)-induced cytotoxicity and reduced expression of pro-inflammatory proteins including cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and tumor necrosis factor-α (TNF- α). Moreover, CFB, CFE and CLS effectively suppressed UVB-induced production of pro-inflammatory mediators such as prostaglandin E₂ (PGE₂) and nitric oxide (NO). In another experiment, topical application of CFB, CFE or CLS prior to UVB irradiation (200 mJ/cm²) on BALB/c mice, inhibited the UVB-elevated protein levels of COX-2, iNOS, and TNF-α. Furthermore, CFB, CFE and CLS suppressed oxidative damages caused by UVB irradiation for example lipid peroxidation and/or protein carbonylation, which seemed to be mediated by up-regulation of antioxidant defense enzymes. These results suggest that C. fragile could be an effective therapeutic agent providing protection against UVB-induced inflammatory and oxidative skin damages.     

Anti-ulcer constituents of Annona squamosa twigs

Phytochemical investigation of Annona squamosa twigs, resulted in isolation and identification of twelve known (1-12) compounds among them one 1-(4-β-D-glucopyranosyloxyphenyl)-2-(β-D-glucopyranosyloxy)-ethane (11) is synthetically known but first time isolated from natural sources. Their structures were elucidated using 1D and 2D NMR spectroscopic analysis. The isolated compounds (2-8, 11) were evaluated for H⁺ K⁺-ATPase activity. Three of these compounds (+)-O-methylarmepavine (2), N-methylcorydaldine (3), isocorydine (6) showed promising anti-secretory activity. Activity of these compounds, comparable to the standard drug omeprazole is novel to our finding. Moreover, there is no information accessible regarding the pharmacological effect of A. squamosa on the gastrointestinal system. This study is the first of its kind to show the significant anti-ulcer effect of A. squamosa. The present study aimed to evaluate the gastroprotective effect of A. squamosa (AS) and to identify its active constituents. Anti-ulcer activity was evaluated against cold restraint (CRU), pyloric ligation (PL), aspirin (ASP), alcohol (AL) induced gastric ulcer and histamine (HA) induced duodenal ulcer model and further confirmed through in vitro assay of H⁺ K⁺-ATPase activity and plasma gastrin level. AS and its chloroform and hexane fraction attenuated ulcer formation in CRU, PL, HA model and displayed anti-secretory activity in vivo through reduced free, total acidity and pepsin in PL, confirmed by in vitro inhibition of H⁺ K⁺-ATPase activity with corresponding decrease in plasma gastrin level. Cytoprotection of AS was apparent with protection in AL, ASP models and enhanced mucin level in PL.     

宽叶杜香叶柄再生体系建立及种质离体保存研究

The tender leafstalks of Ledum palustre var.dilatatum were used as explants for the experiment.Uniform design for the most suitable media for shoots regeneration immediately at base of tender leafstalks,rooting and germplasm conservation in vitro was screened.The results showed that N6+ZT 2.65 mg·L-1+IAA 0.05 mg·L-1 was fits for shoots regeneration,the frequency of shoots induction was higher than 92.5%;MS(modified)+IAA 0.1 mg·L-1+Kt 0.75 mg·L-1 for rooting,the rate of rooting was 98%;N-68+B9 2.5 mg·L-1+ Ph...     

Inhibition of glucose intestinal absorption by kaempferol 3-O-α-rhamnoside purified from Bauhinia megalandra leaves

Glucose intestinal absorption (GIA) is one of the factors that increase glycemia. Its reduction could be an important factor in decreasing hyperglycemia in diabetic patients. It has been shown that the aqueous extract of Bauhinia megalandra leaves inhibits GIA. In the present study we identified a compound present in the extract of B. megalandra responsible for the biological effect. The methanol extract of B. megalandra leaves was fractionated using different solvents, and high-speed counter-current chromatography yielding two pure compounds identified by ¹H NMR and ¹³C NMR as kaempferol 3-O-α-rhamnoside and quercetin 3-O-α-rhamnoside. The first one increased the K_M without changes in the V_MAX of GIA. In addition it exerted an additive inhibitory effect, on GIA, when combined with phlorizin. We suggest that kaempferol 3-O-α-rhamnoside is a competitive inhibitor of intestinal SGLT1 cotransporter.     

Reactive oxygen species scavenging activities and inhibition on DNA oxidative damage of dimeric compounds from the oxidation of (−)-epigallocatechin-3-O-gallate

The dimeric catechins dehydrotheasinensin A (2) and theacitrin C (3) were prepared from the oxidation of (−)-epigallocatechin-3-O-gallate (EGCG, 1), and their antioxidant activity was investigated using a chemiluminescence (CL) method in vitro. Both compounds showed significant inhibitory effects on reactive oxygen species (O₂⁻, H₂O₂ and •OH) and DNA oxidative damage, with 2 being more potent than 3 and EGCG itself.     

Transdermal behaviors comparisons among Evodia rutaecarpa extracts with different purity of evodiamine and rutaecarpine and the effect of topical formulation in vivo

Aim

Evodiamine (EVO) and rutaecapine (RUT), the major active components from Evodia rutaecarpa extract (EE), are recognized as a depended analgesic agent. This study was designed to investigate the effect of purity and chemical enhancers on the transdermal behavior of EVO and RUT, and the pharmacological effect of their topical cream in vivo.

Material and methods

Transdermal delivery across a full thickness pig abdominal skin was detected in vitro by Franz-type diffusion cell, with HPLC for quantification of the permeation of EVO and RUT. The activity of topical cream in vivo was evaluated by a mice pain model induced by formalin and hot plate.

Results

Transdermal characters of EVO and RUT showed a low transdermal rate, long lag time and low cumulative amount. The transdermal rate and cumulative amount could be promoted by lipophilic enhancers, whereas lag time was shortened by hydrophilic surfactant, but these permeation parameters were not markedly influenced by purity of EE (p > 0.05). The effect in vivo was confirmed by analgesic models in topical cream of EE, which produced a significant (p < 0.05) inhibitory effects on pain response in dose-dependent manner.

Conclusion

The purity of EVO and RUT from EE has no significant effect on their permeation through porcine skin, but oleic acid or nerolidol can markedly elevate the transdermal rate of EVO and RUT. High purity of EE is the best choice for topical preparation to increase the drug loading. The effect of EE in vivo is verified by formalin model and hot plate test.     

Kujigamberol, a new dinorlabdane diterpenoid isolated from 85million years old Kuji amber using a biotechnological assay

A new compound, 15,20-dinor-5,7,9-labdatriene-18-ol (1), named kujigamberol, was isolated from amber, fossilized tree resin from the Kuji area in Japan, has been dated as being 85 million years old (late Cretaceous). Kujigamberol was identified using the hypersensitive mutant yeast (zds1? erg3? pdr1? pdr3?) with respect to Ca²⁺-signal transduction. The structure was elucidated on the basis of spectroscopic analysis including 1D NMR, 2D NMR and HR-EI-MS. It was different from known diterpenoids with a similar activity isolated from Baltic amber (agathic acid 15-monomethyl ester (2), dehydroabietic acid (3) and pimaric acid (4)). Kujigamberol showed glycogen synthase kinase-3β (GSK-3β) inhibition activity involving the growth restored activity against the mutant yeast and was cytotoxic to HL60 cells (IC₅₀ = 19.6 μM).     

Inhibition of dipeptidyl peptidase activity by flavonol glycosides of guava (Psidium guajava L.): A key to the beneficial effects of guava in type II diabetes mellitus

Based on the traditional use in popular medicine, the effect of extracts from Psidium guajava L. leaves and of the main flavonol-glycoside components on dipeptidyl-peptidase IV (DP-IV), a key enzyme of blood glucose homoeostasis, has been investigated in-vitro. An ethanolic extract was prepared from dried, powdered leaves of guava and was found to contain seven main flavonol-glycosides, which were isolated by semipreparative HPLC and tested individually. The ethanolic guava leave extract was shown to exert a dose-dependent inhibition of DP-IV, with an IC₅₀ of 380 μg/ml test assay solution. Also the individual flavonol-glycosides inhibited DP-IV dose-dependently, with variations of the effects by a factor of 10, and an overall effect accounting for 100% of that observed for the total guava extract. The recovery of individual flavonol-glycosides in CaCo-2 epithelial cells, a model of gastrointestinal tract absorption, amounted to 2.3–5.3% of the amount available for absorption over 60 min at 37 °C.     

Two new α-pinene derivatives from Angelica sinensis and their anticoagulative activities

Two new α-pinene derivatives (1-2) were isolated from the aerial parts of Angelica sinensis. Their structures were determined by spectroscopic and chemical methods to be 6β,9-dihydroxy-(+)-α-pinene (1) and 9-hydroxy-(+)-α-pinene-6β-O-D-glucoside (2). In the anticoagulative assay, compounds 1 and 2 exhibited weak antithrombin activity and strong antiplatelet aggregation activity in vitro.     

3-Prenyl luteolin,a new prenylated flavone with melanin biosynthesis inhibitory activity from wood of Artocarpus heterophyllus

In our efforts to find new whitening agent from natural resources, we focused on wood of Artocarpus heterophyllus which shows anti-melanogenesis activity. By activity-guided fractionation of A. heterophyllus wood extract, a new prenylated flavonoid, 3-prenyl luteolin (1) was isolated. The IC₅₀ of mushroom tyrosinase inhibitotry activity of 1 was 76.3 µM. The results of the comparison with that of luteolin showed the prenyl substituent at C-3 position of 1 play an important role for revealing tyrosinase inhibition. In melanin formation inhibition on B16 melanoma cells, IC₅₀ of 1 was 56.7 µM with less cytotoxicity.     

白花泡桐优树试管嫁接幼化及组培快繁技术研究

以白花泡桐优树‘白优2号’为试验材料,通过组织培养和试管嫁接方法,对白花泡桐优树材料的幼化技术进行了研究.结果表明:外植体初代培养萌发的嫩芽为最适合的接穗;‘建始桐3号’为试管嫁接较合适的砧木;采用劈接进行;MS+ NAA0.3 mg·L-1+蔗糖30 g·L-1为试管嫁接培养基;继代增殖和生根培养基分别为1/2MS ...     

Effect of Citrus bergamia juice on human neuroblastoma cells in vitro and in metastatic xenograft models

Neuroblastoma is the most common extracranial pediatric solid tumor with poor prognosis in children with disseminated stage of disease. A number of studies show that molecules largely distributed in commonly consumed fruits and vegetables may have anti-tumor activity. In this study we evaluate the effect of Citrus bergamia (bergamot) juice (BJ) in vitro and in a spontaneous metastatic neuroblastoma SCID mouse model. Qualitative and quantitative characterizations of BJ flavonoid fractions were performed by RP-HPLC/PDA/MS. We show that BJ significantly affects SK-N-SH and LAN-1 cell proliferation in vitro, but fails to reduce primary tumor weight in vivo. Moreover, BJ reduced cell adhesiveness and invasion of LAN-1 and SK-N-SH cells in vitro and the number of pulmonary metastases under consideration of the number of tumor cells in the blood in mice inoculated with LAN-1 cells in vivo. These effects without any apparent sign of systemic toxicity confirm the potential clinical interest of BJ and lay the basis for further investigation in cancer.     

白蜡虫多糖免疫调节与抑制肿瘤活性

采用环磷酰胺诱导免疫低下小鼠模型,通过测定模型小鼠的炭粒廓清指数、吞噬指数、胸腺指数和脾指数考察白蜡虫多糖的免疫调节能力;对接种S180瘤细胞的小鼠采用口服和注射方式提供白蜡虫多糖,以抑瘤率考察白蜡虫多糖体内抑瘤活性;以生长率为指标考察白蜡虫多糖对人白血病细胞株、人肺癌细胞株、人胃癌细胞株及人肝癌细胞株等4种肿瘤细胞的抑制作用。结果显示:白蜡虫粗多糖(Cwps)和纯化多糖(Pwps)均能明显增加免疫低下小鼠碳粒廓清指数及吞噬指数,但对胸腺指数和脾指数无改善作用;体内抑瘤试验发现,白蜡虫多糖对小鼠体内S180有明显的抑瘤效果,但对离体肿瘤细胞生长却无明显抑制作用。研究结果表明:白蜡虫多糖能增加免疫低下小鼠的非特性免疫能力,具有体内抑制肿瘤作用,但对肿瘤细胞无细胞毒作用,揭示白蜡虫多糖通过提高小鼠免疫能力达到体内抑制肿瘤的功效。