ABSTRACT
Gigantol is a phenolic component of precious Chinese medicine Dendrobii Caulis, which has many pharmacological activities such as prevent tumor and diabetic cataract. This paper aimed to investigate the molecular mechanism of gigantol in transmembrane transport in human lens epithelial cells(HLECs). Immortalized HLECs were cultured in vitro and inoculated in the laser scanning confocal microscopy(LSCM) medium at 5 000 cells/mL. The fluorescence distribution and intensity of gigantol marked by fluorescence in HLECs were observed by LSCM, and the absorption and distribution of gigantol were expressed as fluorescence intensity. The transmembrane transport process of gigantol in HLECs were monitored. The effects of time, temperature, concentration, transport inhibitors, and different cell lines on the transmembrane absorption and transport of gigantol were compared. HLECs were inoculated on climbing plates of 6-well culture plates, and the ultrastructure of HLECs was detected by atomic force microscopy(AFM) during the transmembrane absorption of non-fluorescent labeled gigantol. The results showed that the transmembrane absorption of gigantol was in time and concentration-dependent manners, which was also able to specifically target HLECs. Energy and carrier transport inhibitors reduced gigantol absorption by HLECs. During transmembrane process of gigantol, the membrane surface of HLECs became rougher and presented different degrees of pits, indicating that the transmembrane transport of gigantol was achieved by active absorption of energy and carrier-mediated endocytosis.
Subject(s)
Humans , Lens, Crystalline/pathology , Cataract/prevention & control , Bibenzyls/pharmacology , Epithelial Cells , Cells, Cultured , ApoptosisABSTRACT
Objective: To investigate the chemical constituents of leaves of Dendrobiumofficinale. Methods: Compounds were isolated from the leaves of D.officinale by column chromatography over Sephadex LH-20, MCI GEL CHP-20P, and ODS as well as by preparative HPLC. Their structures were identified by the analysis of their physical and chemical properties and the spectra data of NMR and MS. Results: Twenty-four compounds were isolated from the leaves of the plant, namely 3,4-dihydroxy-5,4'-dimethoxy bibenzyl (1), moscatilin (2), 4,4'-dihydroxy-3,5-dimethoxybibenzyl (3), densiflorol A (4), (S)-3,4,α-trihydroxy-5,4'-dimethoxybibenzyl (5), gigantol (6), dendrocandin U (7), dendrocandin B (8), loliolide (9), (6R,9S)-dihydroxy-megastigma-4,7-dien-3-one-9-O-β-D-glucopyranoside (10), (6R,9S)-9-hydroxy-megastigma-4,7-dien-3-one-9-O-β-D-glucopyranoside (11), (+)-syringaresinol (12), rutin (13), 2-benzothiazolol (14), p-hydroxyacetophenone (15), p-hydroxyl-benzoic acid (16), protocatechuic acid (17), catechol (18), ethyl p-hydroxyhydrocinnamate (19), 1-glycerol linolenate (20), 2-butoxyethyl linolenate (21), palmitic acid (22), octadecadienoic acid-2,3-dihydroxypropyl ester (23), and urticifolene (24). Conclusion: Itis the first report of the occurrence of compounds 10, 11, 14-23 in Orchidaceae family. Compounds 1, 2, 4, and 6are found in D. officinale for the first time.
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@#To study the inhibitory effect of gigantol on proliferation, migration and invasion of human osteosarcoma U20S cells and to explore the mechanism. Methods: After being treated with different concentrations (10, 25, 50, 75, 100, 150 µmol/L) of gigantol for 24 and 48 h, the proliferation of U20S cells was detected by CCK-8 assay. Transwell assay was used to detect the effects of 25 µmol/L and 50 µmol/L gigantol on the migration and invasion abilities of U20S cells. The lipopolysaccharide (LPS) was used to induce inflammatory reaction in U20S cells before gigantol treatment; qPCR and WB were used to detect the mRNA and protein expressions of NF-κB (p65), TNF-α, IL-6 and PRL-3, respectively. Results: Different concentrations of gigantol could all inhibit the proliferation of sarcoma U20S cells at different time (P<0.05 or P<0.01). The 25 µmol/L and 50 µmol/L of gigantol could significantly inhibit the migration and invasion of osteosarcoma U20S cells (all P<0.01); at the same time, it could inhibit the protein expressions of NF-κB, TNF-α, IL-6 and PRL-3 (P<0.05 or P<0.01). After LPS induction, the mRNA and protein expressions of NF-κB, TNF-α, IL-6 and PRL-3 in U20S cells were significantly increased (all P<0.01); however, the consequent treatment with gigantol (25 and 50 µmol/L) reversed the effects of LPS on U20S cells obviously (P<0.05 or P<0.01). Conclusion: Gigantol can inhibit the proliferation, migration and invasion of osteosarcoma U20S cells, and its mechanism may be related to the regulation of NF-κB/PRL-3 signaling pathway.
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@#AIM: To investigate the protective effects of gigantol on aldose reductase(AR)pathway and oxidative stress induced by high glucose in human retinal microvascular endothelial cells(HRMECs).<p>METHODS: HRMECs were cultured and divided into the five groups: normal glucose group, mannitol group,high glucose group, gigantol group and epalrestat group. Cell vitality was detected by the MTS assay, reactive oxygen species(ROS)levels were detected by DCFH-DA fluorescent probe, protein expressions of nuclear factor kappa B(NF-κB)were observed by Western blotting, mRNA levels of AR, hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor(VEGF)were measured by qRT-PCR.<p>RESULTS: DCFH-DA fluorescent probe levels showed that gigantol reduced ROS generation obviously, and protein expressions of NF-κB, mRNA expressions of AR,HIF-1α and VEGF were all decreased. The expression trends of AR and NF-κB in epalrestat group were consistent with gigantol group.<p>CONCLUSION: Gigantol and epalrestat could inhibit the activation of AR/NF-κB pathway, and gigantol also could inhibit high glucose induced oxidative stress and in HRMECs, which may have a protective effect on diabetic retinopathy(DR), and improve angiogenesis.
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Objective To investigate the chemical constituents from Pholidota rupestris. Methods The chemical constituents from the EtOAc soluble fraction of P. rupestris were studied and 13 compounds were isolated by the method of silica gel column chromatography. The structures of these compounds were elucidated by spectral analyses and chemical properties. Results Thirteen compounds were isolated and identified as confusarin (1), ochrone A (2), coelonin (3), lusianthridin (4), batatanin III (5), gigantol (6), thunalbene (7), cyclopholidone (8), cyclopholidonol (9), syringaresinol (10), β-sitosterol (11), stigamast-5-ene-3β,7α-diol (12), and 5α,8α-epidioxy-(22E,24R)-ergosta-6,22-dien-3β-ol (13). Conclusion All compounds are isolated from this plant for the first time. Compounds 1, 2, and 12 have not been recorded in the genus Pholidota.