This study explored the preservation effect of strigolactone analogs on Gastrodia elata tubers and screened out the suitable preservation measures of G. elata to provide a safer and more effective method for its storage and preservation. Fresh G. elata tubers were treated with 7FGR24, 2,4-D isooctyl ester, and maleic hydrazide, respectively. The growth of flower buds, the activities of CAT, and MDA, and the content of gastrodin and p-hydroxybenzyl alcohol were measured to compare the effects of different compounds on the storage and preservation of G. elata. The effects of different storage temperatures on the preservation of 7FGR24 were compared and analyzed. The gibberellin signal transduction receptor gene GeGID1 was cloned, and the effect of 7FGR24 on the expression level of GeGID1 was analyzed by quantitative polymerase chain reaction(qPCR). The toxicity of the G. elata preservative 7FGR24 was analyzed by intragastric administration in mice to evaluate its safety. The results showed that compared with 2,4-D isooctyl ester and maleic hydrazide, 7FGR24 treatment had a significant inhibitory effect on the growth of G. elata flower buds, and the CAT enzyme activity of G. elata was the highest, indicating that its preservation effect was stronger. Different storage temperatures had different effects on the preservation of G. elata, and the preservation effect was the strongest at 5 ℃. The open reading frame(ORF) of GeGID1 gene was 936 bp in length, and its expression level was significantly down-regulated after 7FGR24 treatment, indicating that 7FGR24 may inhibit the growth of flower buds by inhibiting the gibberellin signal of G. elata, thereby exerting a fresh-keeping effect. Feeding preservative 7FGR24 had no significant effect on the behavior and physiology of mice, indicating that it had no obvious toxicity. This study explored the application of the strigolactone analog 7FGR24 in the storage and preservation of G. elata and preliminarily established a method for the storage and preservation of G. elata, laying a foundation for the molecular mechanism of 7FGR24 in the storage and preservation of G. elata.
Animals ; Mice ; Gastrodia ; Gibberellins ; Maleic Hydrazide ; Esters

The genus Rabdosia is famous for the abundance of diverse and novel ent-kaurane diterpenoids. However, only a few ent-kauranoids have been discovered from R. flexicaulis since the investigation on its chemical constituents is not systematic. To find novel bioactive diterpenoids, the ethyl acetate extract of the above ground part of R. flexicaulis in Daofu County, Sichuan Province was obtained by column chromatography. One new compound and five known ones were identified as flexicaulin E(1), forrestin B(2), inf-lexarabdonin D(3), 7α-hydroxydehydroabietic acid(4), 15-hydroxydehydroabietic acid(5), and pomiferin F(6) by spectral techniques. Compounds 1-3 were the ent-kaurane diterpenoids isolated from this species for the first time. Compounds 4-6, aromatic abie-tanoids, were isolated from the genus Rabdosia for the first time.
Diterpenes ; Diterpenes, Kaurane ; Isodon/chemistry* ; Molecular Structure ; Plant Extracts/chemistry*

Phytohormones play an important role at all stages of plant growth, influencing plant growth and development and regulating plant secondary metabolism, such as the synthesis of flavone, flavonol, anthocyanin, and other flavonoids. Flavonoids, a group of important secondary metabolites ubiquitous in plants, have antioxidative, anti-microbial, and anti-inflammatory activities and thus have a wide range of potential applications in Chinese medicine and food nutrition. With the development of biotechnology, phytohormones' regulation on flavonoids has become a research focus in recent years. This study reviewed the research progress on the mechanism of common phytohormones, such as abscisic acid, gibberellin, methyl jasmonate, and salicylic acid, in regulating flavonoid metabolism, and discussed the molecular mechanism of the synthesis and accumulation of flavonoids, aiming at clarifying the key role of phytohormones in modulating flavonoid metabolism. The result is of guiding significance for improving the content of flavonoids in plants through rational use of phytohormones and of reference value for exploring the mechanism of hormones in regulating flavonoid metabolism.
Abscisic Acid ; Flavonoids ; Gene Expression Regulation, Plant ; Gibberellins ; Plant Development ; Plant Growth Regulators

To prepare and optimize the self-microemulsion co-loaded with tenuifolin and β-asarone(TF/ASA-SMEDDS) and evaluate its quality. The prescription compositions of TF/ASA-SMEDDS were screened by solubility test, single factor test and pseudo-tern-ary phase diagram, and the prescriptions were further optimized by Box-Behnken response surface method, with the drug loading and particle size as the evaluation indexes. Then the optimized TF/ASA-SMEDDS was evaluated for emulsified appearance, particle size, morphology and drug release in vitro. The optimized prescription for TF/ASA-SMEDDS was as follows: caprylic citrate triglyceride polyoxyethylene castor oil-glycerol(10.8∶39.2∶50), drug loading of(5.563±0.065) mg·g~(-1) for tenuifolin and(5.526±0.022) mg·g~(-1) for β-asarone; uniform and transparent pan-blue nanoemulsion can be formed after emulsification, with particle size of(28.84±0.44) nm. TEM showed that TF/ASA-SMEDDS can form spherical droplets with a uniform particle size after emulsification; In vitro release test results showed that the drug release rate and cumulative release of tenuifolin and β-asarone were significantly improved. The preparation process of TF/ASA-SMEDDS was simple and can effectively improve in vitro release of tenuifolin and β-asarone.
Anisoles ; Biological Availability ; Diterpenes, Kaurane ; Drug Delivery Systems ; Emulsions ; Particle Size ; Solubility ; Surface-Active Agents

Seed vigor is a key factor affecting seed quality. The mechanical drying process exerts a significant influence on rice seed vigor. The initial moisture content (IMC) and drying temperature are considered the main factors affecting rice seed vigor through mechanical drying. This study aimed to determine the optimum drying temperature for rice seeds according to the IMC, and elucidate the mechanisms mediating the effects of drying temperature and IMC on seed vigor. Rice seeds with three different IMCs (20%, 25%, and 30%) were dried to the target moisture content (14%) at four different drying temperatures. The results showed that the drying temperature and IMC had significant effects on the drying performance and vigor of the rice seeds. The upper limits of drying temperature for rice seeds with 20%, 25%, and 30% IMCs were 45, 42, and 38 °C, respectively. The drying rate and seed temperature increased significantly with increasing drying temperature. The drying temperature, drying rate, and seed temperature showed extremely significant negative correlations with germination energy (GE), germination rate, germination index (GI), and vigor index (VI). A high IMC and drying temperature probably induced a massive accumulation of hydrogen peroxide (H₂O₂) and superoxide anions in the seeds, enhanced superoxide dismutase (SOD) and catalase (CAT) activity, and increased the abscisic acid (ABA) content. In the early stage of seed germination, the IMC and drying temperature regulated seed germination through the metabolism of H₂O₂, gibberellin acid (GA), ABA, and α-amylase. These results indicate that the metabolism of reactive oxygen species (ROS), antioxidant enzymes, GA, ABA, and α-amylase might be involved in the mediation of the effects of drying temperature on seed vigor. The results of this study provide a theoretical basis and technical guidance for the mechanical drying of rice seeds.
Abscisic Acid/metabolism* ; Antioxidants/pharmacology* ; Catalase/metabolism* ; Gene Expression Regulation, Plant/drug effects* ; Germination ; Gibberellins/metabolism* ; Hydrogen Peroxide/chemistry* ; Malondialdehyde/chemistry* ; Oryza/metabolism* ; Oxygen/chemistry* ; Plant Proteins/genetics* ; Reactive Oxygen Species ; Seeds/metabolism* ; Superoxide Dismutase/metabolism* ; Superoxides/chemistry* ; Temperature ; Weather ; alpha-Amylases/metabolism*

OBJECTIVE: To investigate the effects of oridonin (ORI) on the proliferation and apoptosis of human multiple myeloma cell line H929 and its possible mechanism. METHODS: H929 cells were exposed to ORI 0、4、8、12、16、20、24、28、32 μmol/L for 12, 24 and 36 hours respectively. The prolifcration inhibitory effect of ORI on H929 cells was determined by MTT assay and then the working concentrations of ORI were determined. The morphological changes and apoptosis of H929 cells were observed by TUNEL (TdT-mediated dUTP Nick-End Labeling) and fluorescence microscopy. The apoptosis rate of H929 cells was detected by flow cytometry with Annexin V-FITC/PI staining. The protein expressions of pro-caspase-3, BCL-2，p-PI3K, p-Akt, BAX, Cleaved PARP and p-JNK, p-ERK and p-p38 in H929 cells were detected by Western blot. RESULTS: Compared with the control group, the proliferation of H929 cells treated with the ORI of 8-16 μmol/L was significantly inhibited and the apoptosis of H929 cells was obviously increased in dose- and time-dependent manners. As for morphological changes, the characteristics of apoptotic cells were presented in H929 cells treated with ORI for 24 hours. The protein levels of pro-caspase-3, BCL-2，p-PI3K, p-Akt were down-regulated with increasing of ORI concentration（r=0.9861, r=0.9725, r=0.9413, r=0.9373）, while the BAX, Cleaved PARP and p-JNK, p-ERK and p-p38 were up-regulated（r=0.9178, r=0.8877, r=0.882, r=0.9645, r=0.8623）. CONCLUSION The ORI possesses anti-myeloma effects, can inhibit the proliferation and induce the apoptosis of H929 cell line in vitro. Its potential mechanism may be related with up-regulating the MAPK and down-regulating the PI3K/Akt signal pathways.
Apoptosis ; Caspase 3 ; Cell Line, Tumor ; Cell Proliferation ; Diterpenes, Kaurane ; Humans ; Multiple Myeloma ; Phosphatidylinositol 3-Kinases

OBJECTIVE: To investigate the influence of oridonin on the killing activity of NK-92 MI cells targeting THP1 and the related mechanism. METHODS: The killing activity of NK-92 MI to THP1 before and after oridonin treatment was detected by LDH release assay; the expression of natural killer cell ligands activating receptor D (NKG2D, including MICA, MICB, ULBP1, ULBP2 and ULBP3) was detected by real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot respectively; the expression of cytokine TNF-α, TNF-β and IFN-γ in the co-culture supernatant of NK-92 MI cells and THP1 cells were measured by ELISA. RESULTS: The killing efficiency after oridonin treatment at different effector-target ratio (1:1, 5:1, 10:1) was all significantly up-regulated in comparison with that before oridonin treatment (P＜0.05). QRT-PCR and Western blot showed that the expressions of mRNA and protein levels of MICB, ULBP1, ULBP2 increased to varying degree (P＜0.05), but the expression levels of MICA and ULBP3 were not statistically significant between experimental group and control group (P＞0.05). ELISA results indicated that IFN-γ and TNF-β release were significantly increased after oridonin treatment (P＜0.05), however, the TNF-α release was not statistically different in comparison with control group (P＞0.05). CONCLUSION Oridonin can significantly improve killing efficiency of NK-92 MI on THP1, that might be related with up-regulation of MICB, ULBP1 and ULBP2 expression and promotion of IFN-γ and TNF-β release.
Cell Line, Tumor ; Diterpenes, Kaurane ; pharmacology ; GPI-Linked Proteins ; Histocompatibility Antigens Class I ; Humans

OBJECTIVE: To investigate the differentiation of acute promyelocytic leukemia (APL) cells induced by adenosine targeting Prx III. METHODS: HL-60 cells were divided into four groups: control group, all-trans retinoic acid (ATRA) group, adenanthin group and ATRA+adenanthin group. Cell morphologic changes were observed under optical microscope. The influence of adenanthin on the differentiation of HL-60 was observed by nitro blue tetrazolium chloride (NBT) test. Cell surface differentiation antigens CD11b expression was measured by flow cytometry. The protein expression of Prx III was detected by immunohistochemical assay. RESULTS: Adenanthin could induce the differentiation of HL-60 cells; the NBT reduction positive rate in ATRA+adenanthin group was significantly higher than that in ATRA group and adenanthin group (P＜0.05). The percentage of CD11b positive cells in ATRA+adenanthin group (43.62％±1.38％) was higher than that in adenanthin group (28.15％±1.78％), ATRA group (36.72％±1.33％) and control group (7.99％±1.78％) (P＜0. 05). The content of Prx Ⅲ protein in adenanthin group was significantly higher than that in control group and ATRA group (P＜0.05). CONCLUSION Adenanthin and ATRA have a synergistic effect on the differentiation and maturation of HL-60 cells, and its mechanism may be related with regulation of Prx III expression.
Cell Differentiation ; Diterpenes, Kaurane ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; Peroxiredoxin III ; Tretinoin

Gibberellin is an essential plant hormone that plays an important regulatory role throughout the life cycle of higher plants. A total of 23 genes involved in gibberellin action were identified from Phyllostachys edulis genome, including 8 GA20ox and 1 GA3ox genes involved in the gibberellin biosynthesis, 8 GA2ox genes involved in the metabolism of gibberellin, 2 GID1 genes involved in gibberellin perception, 2 GID2 genes and 2 DELLA genes involved in gibberellin signal transduction. Phylogenetic analysis of these genes from Arabidopsis, Oryza sativa and Phyllostachys edulis revealed that gibberellin biosynthesis, metabolism, and signaling pathways are conserved in these species. Treatment of seeds and seedlings of bamboo with exogenous gibberellin revealed that gibberellin significantly increased seed germination rate and stem elongation of seedlings, and had the best concentration of action. The expression levels of GA20ox and GA3ox genes in the bamboo seedlings were down-regulated and the expression of the active gibberellin-degrading gene GA2ox was up-regulated after GA3 treatment, and the transcriptional level of the gibberellin receptor GID1 and the positive regulatory gene GID2 was significantly increased while the expression of the negative regulatory gene DELLA was decreased. These genes have significant differences in the expression of different spatial locations of bamboo shoot stems, GA20ox, GA3ox, GA2ox, GID1 and GID2 are all expressed in the upper part of bamboo shoots, while the repressor gene DELLA accumulates at the bottom of the shoots and is hardly expressed at the top.
Arabidopsis ; Gene Expression Regulation, Plant ; Gibberellins ; biosynthesis ; Phylogeny ; Plant Growth Regulators ; Plant Proteins ; Poaceae

Nine new ent-kaurane diterpenoids, named scopariusols L-T (1-9), were isolated from the aerial parts of Isodon scoparius. Their structures were characterized mainly by analyzing the NMR and HR-ESI-MS data, and the absolute configuration of 1 was determined by single-crystal X-ray diffraction. Compound 1 was active against five human tumor cell lines (HL-60, SMMC-7721, A-549, MCF-7, and SW-480), and it also inhibited NO production in LPS-stimulated RAW264.7 cells, with an IC value of 0.6 μmol·L.
Animals ; Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Crystallography, X-Ray ; Diterpenes, Kaurane ; chemistry ; isolation & purification ; pharmacology ; Drug Screening Assays, Antitumor ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; HL-60 Cells ; Humans ; Isodon ; chemistry ; Lipopolysaccharides ; pharmacology ; Macrophages ; drug effects ; Mice ; Molecular Structure ; Nitric Oxide ; biosynthesis ; Nuclear Magnetic Resonance, Biomolecular ; Plant Components, Aerial ; chemistry ; RAW 264.7 Cells