1.Preparation of calcium phosphate nanoflowers and evaluation of their antioxidant and osteogenic induction capabilities in vitro.
Mingyu JIA ; Zhihong CHEN ; Huajian ZHOU ; Yukang ZHANG ; Min WU
Chinese Journal of Reparative and Reconstructive Surgery 2025;39(9):1203-1211
OBJECTIVE:
To investigate the antioxidant and osteogenic induction capabilities of calcium phosphate nanoflowers (hereinafter referred to as nanoflowers) in vitro at different concentrations.
METHODS:
Nanoflowers were prepared using gelatin, tripolyphosphate, and calcium chloride. Their morphology, microstructure, elemental composition and distribution, diameter, and molecular constitution were characterized using scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, and energy-dispersive spectroscopy. Femurs and tibias were harvested from twelve 4-week-old Sprague Dawley rats, and bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured using the whole bone marrow adherent method, followed by passaging. The third passage cells were identified as stem cells by flow cytometry and then co-cultured with nanoflowers at concentrations of 0, 0.4, 0.8, 1.2, 1.6, 2.0, 2.4, 2.8, 3.2, and 3.6 mg/mL. Cell counting kit 8 (CCK-8) assay was performed to screen for the optimal concentration that demonstrated the best cell viability, which was subsequently used as the experimental concentration for further studies. After co-culturing BMSCs with the screened concentration of nanoflowers, the biocompatibility of the nanoflowers was verified through live/dead cell staining, scratch assay, and cytoskeleton staining. The antioxidant capacity was assessed by using reactive oxygen species (ROS) fluorescence staining. The in vitro osteoinductive ability was evaluated via alkaline phosphatase (ALP) staining, alizarin red staining, and immunofluorescence staining of osteocalcin (OCN) and Runt-related transcription factor 2 (RUNX2). All the above indicators were compared with the control group of normally cultured BMSCs without the addition of nanoflowers.
RESULTS:
Scanning electron microscopy revealed that the prepared nanoflowers exhibited a flower-like structure; transmission electron microscopy scans discovered that the nanoflowers possessed a multi-layered structure, and high-magnification images displayed continuous atomic arrangements, with the nanoflower diameter measuring (2.00±0.25) μm; energy-dispersive spectroscopy indicated that the nanoflowers contained elements such as C, N, O, P, and Ca, which were uniformly distributed across the flower region; Fourier transform infrared spectroscopy analyzed the absorption peaks of each component, demonstrating the successful preparation of the nanoflowers. Through CCK-8 screening, the concentrations of 0.8, 1.2, and 1.6 mg/mL were selected for subsequent experiments. The live/dead cell staining showed that nanoflowers at different concentrations exhibited good cell compatibility, with the 1.2 mg/mL concentration being the best (P<0.05). The scratch assay results indicated that the cell migration ability in the 1.2 mg/mL group was superior to the other groups (P<0.05). The cytoskeleton staining revealed that the cell morphology was well-extended in all concentration groups, with no significant difference compared to the control group. The ROS fluorescence staining demonstrated that the ROS fluorescence in all concentration groups decreased compared to the control group after lipopolysaccharide induction (P<0.05), with the 1.2 mg/mL group showing the weakest fluorescence. The ALP staining showed blue-purple nodular deposits around the cells in all groups, with the 1.2 mg/mL group being significantly more prominent. The alizarin red staining displayed orange-red mineralized nodules around the cells in all groups, with the 1.2 mg/mL group having more and denser nodules. The immunofluorescence staining revealed that the expressions of RUNX2 and OCN proteins in all concentration groups increased compared to the control group, with the 1.2 mg/mL group showing the strongest protein expression (P<0.05).
CONCLUSION
The study successfully prepares nanoflowers, among which the 1.2 mg/mL nanoflowers exhibits excellent cell compatibility, antioxidant properties, and osteogenic induction capability, demonstrating their potential as an artificial bone substitute material.
Animals
;
Osteogenesis/drug effects*
;
Mesenchymal Stem Cells/drug effects*
;
Calcium Phosphates/pharmacology*
;
Rats, Sprague-Dawley
;
Rats
;
Antioxidants/chemistry*
;
Cells, Cultured
;
Cell Differentiation/drug effects*
;
Nanostructures/chemistry*
;
Tissue Engineering/methods*
;
Bone Marrow Cells/cytology*
;
Coculture Techniques
;
Tissue Scaffolds/chemistry*
;
Male
;
Biocompatible Materials/chemistry*
;
Cell Survival
;
Core Binding Factor Alpha 1 Subunit/metabolism*
;
Cell Proliferation
2.Cytoprotective activity of Pogonatherum paniceum (Lam.) Hack. ethanolic extract evaluated by synchrotron radiation-based Fourier transform infrared microspectroscopy.
Benjawan DUNKHUNTHOD ; Kanjana THUMANU ; Yothin TEETHAISONG ; Priyada SITTISART ; Patcharawan SITTISART
Journal of Integrative Medicine 2025;23(2):182-194
OBJECTIVE:
The present study investigated the cytoprotective effects of a Pogonatherum paniceum extract prepared with 80% ethanol (PPE) using synchrotron radiation-based Fourier transform infrared (SR-FTIR) microspectroscopy and determined its phytochemical profile.
METHODS:
The volatile and polyphenolic compounds in PPE were characterized using gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry, respectively. The antioxidant capacity of PPE was evaluated using chemical and cell-based assays. The SR-FTIR microspectroscopy was performed to evaluate the cytoprotective effect of PPE by identifying changes in macromolecule composition in tert-butyl hydroperoxide (tBuOOH)-induced oxidative damage in RAW264.7 cells.
RESULTS:
A total of 48 volatile compounds and 28 polyphenol components were found in PPE. PPE exhibited a high potential for antioxidant activity by scavenging the intracellular reactive oxygen species in tBuOOH-induced oxidative damage in RAW264.7 cells. PPE treatment also significantly protected RAW264.7 cells against tBuOOH-induced toxicity and restored cell viability. The SR-FTIR analysis revealed that tBuOOH increased the lipid and ester lipid content in RAW264.7 cells. The PPE exerted a cytoprotective effect by decreasing the levels of lipid and ester lipid compounds that had been elevated by tBuOOH in RAW264.7 cells. These findings indicate that PPE has cytoprotective potential due to its ability to inhibit endogenous reactive oxygen species.
CONCLUSION
This study extends the current knowledge on the phytochemistry of PPE and its antioxidant and cytoprotective effects. These findings support the use of SR-FTIR microspectroscopy to determine the cytoprotective effects of natural products. PPE extract may be a candidate compound for new therapeutics and nutraceuticals that target the prevention of oxidative stress-associated diseases. Please cite this article as: Dunkhunthod B, Thumanu K, Teethaisong Y, Sittisart P, Sittisart P. Cytoprotective activity of Pogonatherum paniceum (Lam.) Hack. ethanolic extract evaluated by synchrotron radiation-based Fourier transform infrared microspectroscopy. J Integr Med. 2025; 23(2): 182-194.
Animals
;
Mice
;
Spectroscopy, Fourier Transform Infrared/methods*
;
Plant Extracts/chemistry*
;
RAW 264.7 Cells
;
Synchrotrons
;
Oxidative Stress/drug effects*
;
Antioxidants/pharmacology*
;
Ethanol/chemistry*
;
Poaceae/chemistry*
;
Cell Survival/drug effects*
;
Cytoprotection/drug effects*
;
Reactive Oxygen Species/metabolism*
;
tert-Butylhydroperoxide
3.Research progress in biological activities and oocyte aging-regulating effect of EGCG.
Weiying ZHANG ; Huizhu ZHANG ; Yujun LI ; Daoliang LAN ; Xianrong XIONG ; Yaying WANG ; Jian LI ; Honghong HE
Chinese Journal of Biotechnology 2024;40(12):4382-4395
Epigallocatechin gallate (EGCG), the predominant polyphenol in green tea, exerts a spectrum of physiological activities, including antioxidant, anticancer, and anti-inflammatory effects. Emerging research underscores the significance of EGCG in modulating oocyte aging. EGCG can enhance antioxidant defenses, improve mitochondrial functions, and inhibit apoptotic pathways, thereby retarding the aging of oocytes. This review delineates the main molecular features of EGCG and expounds its regulatory mechanisms concerning oocyte aging, enriching the knowledge on the role of EGCG in the amelioration of oocyte aging.
Catechin/pharmacology*
;
Oocytes/metabolism*
;
Humans
;
Animals
;
Antioxidants/pharmacology*
;
Female
;
Cellular Senescence/drug effects*
;
Tea/chemistry*
;
Apoptosis/drug effects*
4.Lyciumbarbarum polysaccharides ameliorate canine acute liver injury by reducing oxidative stress, protecting mitochondrial function, and regulating metabolic pathways.
Jianjia HUANG ; Yuman BAI ; Wenting XIE ; Rongmei WANG ; Wenyue QIU ; Shuilian ZHOU ; Zhaoxin TANG ; Jianzhao LIAO ; Rongsheng SU
Journal of Zhejiang University. Science. B 2023;24(2):157-171
The development of acute liver injury can result in liver cirrhosis, liver failure, and even liver cancer, yet there is currently no effective therapy for it. The purpose of this study was to investigate the protective effect and therapeutic mechanism of Lyciumbarbarum polysaccharides (LBPs) on acute liver injury induced by carbon tetrachloride (CCl4). To create a model of acute liver injury, experimental canines received an intraperitoneal injection of 1 mL/kg of CCl4 solution. The experimental canines in the therapy group were then fed LBPs (20 mg/kg). CCl4-induced liver structural damage, excessive fibrosis, and reduced mitochondrial density were all improved by LBPs, according to microstructure data. By suppressing Kelch-like epichlorohydrin (ECH)-associated protein 1 (Keap1), promoting the production of sequestosome 1 (SQSTM1)/p62, nuclear factor erythroid 2-related factor 2 (Nrf2), and phase II detoxification genes and proteins downstream of Nrf2, and restoring the activity of anti-oxidant enzymes like catalase (CAT), LBPs can restore and increase the antioxidant capacity of liver. To lessen mitochondrial damage, LBPs can also enhance mitochondrial respiration, raise tissue adenosine triphosphate (ATP) levels, and reactivate the respiratory chain complexes I‒V. According to serum metabolomics, the therapeutic impact of LBPs on acute liver damage is accomplished mostly by controlling the pathways to lipid metabolism. 9-Hydroxyoctadecadienoic acid (9-HODE), lysophosphatidylcholine (LysoPC/LPC), and phosphatidylethanolamine (PE) may be potential indicators of acute liver injury. This study confirmed that LBPs, an effective hepatoprotective drug, may cure acute liver injury by lowering oxidative stress, repairing mitochondrial damage, and regulating metabolic pathways.
Animals
;
Dogs
;
Antioxidants/metabolism*
;
Carbon Tetrachloride
;
Chemical and Drug Induced Liver Injury/drug therapy*
;
Kelch-Like ECH-Associated Protein 1/metabolism*
;
Liver
;
Metabolic Networks and Pathways
;
Mitochondria/metabolism*
;
NF-E2-Related Factor 2/metabolism*
;
Oxidative Stress
;
Polysaccharides/pharmacology*
;
Lycium/chemistry*
5.Anti-oxidant and Anti-inflammatory Effects of Ethanol Extract from Polygala sibirica L. var megalopha Fr. on Lipopolysaccharide-Stimulated RAW264.7 Cells.
Cheng-Liu YANG ; Shi-Bo WANG ; Wen-Ping HE ; Jin-Juan LIU
Chinese journal of integrative medicine 2023;29(10):905-913
OBJECTIVE:
To investigate the anti-oxidant and anti-inflammatory effects of ethanol extract of Polygala sibirica L. var megalopha Fr. (EEP) on RAW264.7 mouse macrophages.
METHODS:
RAW264.7 cells were pretreated with 0-200 µg/mL EEP or vehicle for 2 h prior to exposure to 1 µg/mL lipopolysaccharide (LPS) for 24 h. Nitric oxide (NO) and prostaglandin (PGE2) production were determined by Griess reagent and enzyme-linked immunosorbent assay (ELISA), respectively. The mRNA levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor α (TNF-α), interleukin-1beta (IL-1β), and IL-6 were determined using reverse transcription polymerase chain reaction (RT-PCR). Western blot assay was used to determine the protein expressions of iNOS, COX-2, phosphorylation of extracellular regulated protein kinases (ERK1/2), c-Jun N-terminal kinase (JNK), inhibitory subunit of nuclear factor Kappa B alpha (Iκ B-α) and p38. Immunofluorescence was used to observe the nuclear expression of nuclear factor-κ B p65 (NF-κ B p65). Additionally, the anti-oxidant potential of EEP was evaluated by reactive oxygen species (ROS) production and the activities of catalase (CAT) and superoxide dismutase (SOD). The 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (OH), superoxide anion (O2-) radical and nitrite scavenging activity were also measured.
RESULTS:
The total polyphenol and flavonoid contents of EEP were 23.50±2.16 mg gallic acid equivalent/100 g and 43.78±3.81 mg rutin equivalent/100 g. With EEP treatment (100 and 150 µg/mL), there was a notable decrease in NO and PGE2 production induced by LPS in RAW264.7 cells by downregulation of iNOS and COX-2 mRNA and protein expressions (P<0.01 or P<0.05). Furthermore, with EEP treatment (150 µg/mL), there was a decrease in the mRNA expression levels of TNF-α, IL-1β and IL-6, as well as in the phosphorylation of ERK, JNK and p38 mitogen-activated protein kinase (MAPK, P<0.01 or P<0.05), by blocking the nuclear translocation of NF-κ B p65 in LPS-stimulated cells. In addition, EEP (100 and 150 µg/mL) led to an increase in the anti-oxidant enzymes activity of SOD and CAT, with a concomitant decrease in ROS production (P<0.01 or P<0.05). EEP also indicated the DPPH, OH, O2- radical and nitrite scavenging activity.
CONCLUSION
EEP inhibited inflammatory responses in activated macrophages through blocking MAPK/NF-κ B pathway and protected against oxidative stress.
Animals
;
Mice
;
Antioxidants/pharmacology*
;
Lipopolysaccharides/pharmacology*
;
Polygala
;
Transcription Factor RelA/metabolism*
;
Tumor Necrosis Factor-alpha/metabolism*
;
Ethanol/chemistry*
;
Interleukin-6/metabolism*
;
Anti-Inflammatory Agents/chemistry*
;
Reactive Oxygen Species/metabolism*
;
Cyclooxygenase 2/metabolism*
;
Nitrites/metabolism*
;
NF-kappa B/metabolism*
;
Nitric Oxide/metabolism*
;
Superoxide Dismutase/metabolism*
;
RNA, Messenger
;
Nitric Oxide Synthase Type II/metabolism*
6.Anti-hypoxic pharmacological effects of betelnut polyphenols.
Jianghong MA ; Xing DU ; Anpeng ZHAO ; Zihan WANG ; Qianwen GUO ; Ningning QIN ; Rong WANG
Journal of Central South University(Medical Sciences) 2022;47(4):512-520
Areca catechu L. medicinal materials and their preparations are widely used in clinical practice. Betelnut polyphenol is one of the main chemical components with antioxidant, anti-inflammatory, and antibacterial effects. With continuous increase of high altitude activities, tissue oxidative damage caused by high altitude hypoxia seriously affects the ability to work, and the studies on anti-hypoxia drugs are particularly important. Recent studies have shown that betelnut polyphenols have protective effects on oxidative stress injury caused by hypoxia via improving blood gas index of hypoxic organism, increasing superoxide dismutase glutathione catalase activity, and scavenging excessive free radicals. The effects of betelnut polyphenols against hypoxia and oxidative damage protection suggest that betelnut polyphenols can be used as potential anti-hypoxia drugs and posses clinical prospects.
Antioxidants/pharmacology*
;
Areca/chemistry*
;
Humans
;
Hypoxia
;
Oxidative Stress
;
Polyphenols/pharmacology*
;
Superoxide Dismutase/metabolism*
7.Drying temperature affects rice seed vigor via gibberellin, abscisic acid, and antioxidant enzyme metabolism.
Yu-Tao HUANG ; Wei WU ; Wen-Xiong ZOU ; Hua-Ping WU ; Dong-Dong CAO
Journal of Zhejiang University. Science. B 2020;21(10):796-810
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*
8.Untargeted metabonomics study of ginseng in treatment of spleen-Qi deficiency.
Ni WANG ; Na ZHANG ; Tie LI ; Mei WANG ; Xin HUANG ; Shu-Ying LIU
China Journal of Chinese Materia Medica 2020;45(2):398-404
Ginseng has been used to treat Qi-deficiency syndrome up to now, while the therapeutic mechanism is still unclear. In order to explore the mechanism of ginseng in the treatment of Qi-deficiency constitution, the untargeted metabonomics with blood was studied based on rapid resolution high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(RRLC-Q-TOF-MS). In the results, 13 potential biomarkers were found and identified, which mainly involved in the body's antioxidant and immune functions and energy, glycerol, fatty acid, sugar metabolism and bile acid metabolism. The results of blood biochemical analysis also indicated that ginseng could regulate the body's energy metabolism, immune functions and antioxidant capacity in spleen-Qi deficiency constitution. This study revealed the mechanism of ginseng in the treatment of spleen-Qi deficiency using the blood metabonomics, which could provide technological support and scientific basis for further research on ginseng treatment of Qi-deficiency.
Antioxidants
;
Biomarkers/blood*
;
Chromatography, High Pressure Liquid
;
Drugs, Chinese Herbal/therapeutic use*
;
Energy Metabolism
;
Humans
;
Immune System
;
Medicine, Chinese Traditional
;
Metabolomics
;
Panax/chemistry*
;
Qi
;
Spleen
9.Effects of fungi fraction on growth and anti-oxidative activity of Eleutherococcus senticosus.
Shuang ZHANG ; Sheng-Lei GUO ; Qian-Bo WANG ; Yue LIU ; Hong-Wei SHEN ; Zhen-Yue WANG
China Journal of Chinese Materia Medica 2019;44(8):1517-1523
The present study was conducted to explore the effect of endophytic fungi fraction on growth and anti-oxidative activity of Eleutherococcus senticosus. The growth,yield,contents of MDA,and antioxidant activities were assessed in E. senticosus under five fungi fractions,namely BZ,MH,DT,JS,and XFZ. The results showed that fungi fractions and component significantly affected the growth,low concentration of DT fungi fraction significantly increased the biomass of E. senticosus,reduced the MDA content in cells,and the antioxidant activities of the aqueous extracts were superior to the others. The results indicated that low concentration of DT fungi fraction was the optimum fraction to achieve high yield and quality of E. senticosus.
Antioxidants
;
metabolism
;
Eleutherococcus
;
growth & development
;
metabolism
;
Fungi
;
chemistry
;
Malondialdehyde
;
metabolism
;
Oxidative Stress
10.Evaluation of in vitro antioxidant, antiglycation and antimicrobial potential of indigenous Myanmar medicinal plants.
The Su MOE ; Htet Htet WIN ; Thin Thin HLAING ; War War LWIN ; Zaw Min HTET ; Khin Mar MYA
Journal of Integrative Medicine 2018;16(5):358-366
OBJECTIVEMyanmar has a long history of using medicinal plants for treatment of various diseases. To the best of our knowledge there are no previous reports on antiglycation activities of medicinal plants from Myanmar. Therefore, this study was aimed to evaluate the antioxidant, antiglycation and antimicrobial properties of 20 ethanolic extracts from 17 medicinal plants indigenous to Myanmar.
METHODSIn vitro scavenging assays of 2,2-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide (NO), superoxide (SO) radicals were used to determine the antioxidant activities. Folin-Ciocalteu's method was performed to determine the total phenolic content. Antiglycation and antimicrobial activities were detected by bovine serum albumin-fluorescent assay and agar well diffusion method.
RESULTSTerminalia chebula Retz. (Fruit), containing the highest total phenolic content, showed high antioxidant activities with inhibition of 77.98% ± 0.92%, 88.95% ± 2.42%, 88.56% ± 1.87% and 70.74%± 2.57% for DPPH, NO, SO assays and antiglycation activity respectively. It also showed the antimicrobial activities against Staphylococcus aureus, Bacillus cereus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans with inhibition zone of 19, 18, 17, 25 and 15 mm, respectively. Garcinia mangostana Linn. showed the strongest activities for SO and antiglycation assays with inhibition of 93.68% ± 2.63% and 82.37% ± 1.78%. Bark of Melia sp. was the best NO radical scavenger with inhibition rate of 89.39%± 0.60%.
CONCLUSIONThe results suggest that these plants are potential sources of antioxidants with free radical-scavenging and antiglycation activities and could be useful for decreasing the oxidative stress and glycation end-product formation in glycation-related diseases.
Anti-Bacterial Agents ; analysis ; pharmacology ; Anti-Infective Agents ; analysis ; pharmacology ; Antioxidants ; analysis ; pharmacology ; Bacteria ; drug effects ; growth & development ; Biphenyl Compounds ; metabolism ; Candida albicans ; drug effects ; growth & development ; Fruit ; Garcinia ; chemistry ; Glycation End Products, Advanced ; metabolism ; Humans ; Magnoliopsida ; chemistry ; Medicine, Traditional ; Melia ; chemistry ; Myanmar ; Nitric Oxide ; metabolism ; Oxidative Stress ; drug effects ; Phenols ; analysis ; pharmacology ; Phytotherapy ; Picrates ; metabolism ; Plant Bark ; Plant Extracts ; chemistry ; pharmacology ; Plants, Medicinal ; Superoxides ; Terminalia ; chemistry

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