ABSTRACT
Osteoporosis is the most common osteolytic disease characterized by excessive osteoclast formation and resultant bone loss, which afflicts millions of patients around the world. Astilbin, a traditional herb, is known to have anti-inflammatory, antioxidant and antihepatic properties, but its role in osteoporosis treatment has not yet been confirmed. In our study, astilbin was found to have an inhibitory effect on the RANKL-induced formation and function of OCs in a dose-dependent manner without cytotoxicity. These effects were attributed to its ability to suppress the activity of two transcription factors (NFATc1 and c-Fos) indispensable for osteoclast formation, followed by inhibition of the expression of bone resorption-related genes and proteins (Acp5/TRAcP, CTSK, V-ATPase-d2 and integrin ß3). Furthermore, we examined the underlying mechanisms and found that astilbin repressed osteoclastogenesis by blocking Ca2+ oscillations and the NF-κB and MAPK pathways. In addition, the therapeutic effect of MA on preventing bone loss in vivo was further confirmed in an ovariectomized mouse model. Therefore, considering its ability to inhibit RANKL-mediated osteoclastogenesis and the underlying mechanisms, astilbin might be a potential candidate for treating osteolytic bone diseases.
Subject(s)
Bone Resorption/prevention & control , Flavonols/pharmacology , Osteoclasts/drug effects , Osteogenesis/drug effects , RANK Ligand/pharmacology , Animals , Cells, Cultured , Drugs, Chinese Herbal/pharmacology , Female , Gene Expression/drug effects , Humans , Integrin beta3/genetics , Integrin beta3/metabolism , Mice , Mice, Inbred C57BL , NFATC Transcription Factors/metabolism , Osteoclasts/metabolism , Osteogenesis/genetics , Ovariectomy , Phytotherapy/methods , Proto-Oncogene Proteins c-fos/metabolism , RAW 264.7 Cells , Tartrate-Resistant Acid Phosphatase/genetics , Tartrate-Resistant Acid Phosphatase/metabolismABSTRACT
Osteoporosis is the most common osteolytic disease characterized by excessive osteoclast formation and resultant bone loss, which afflicts millions of patients around the world. Madecassoside (MA), isolated from Centella asiatica, was reported to have anti-inflammatory and antioxidant activities, but its role in osteoporosis treatment has not yet been confirmed. In our study, MA was found to have an inhibitory effect on the RANKL-induced formation and function of OCs in a dose-dependent manner without cytotoxicity. These effects were attributed to its ability to suppress the activity of two transcription factors (NFATc1 and c-Fos) indispensable for osteoclast formation, followed by inhibition of the expression of bone resorption-related genes and proteins (Acp5/TRAcP, CTSK, ATP6V0D2/V-ATPase-d2, and integrin ß3). Furthermore, we examined the underlying mechanisms and found that MA represses osteoclastogenesis by blocking Ca2+ oscillations and the NF-κB and MAPK pathways. In addition, the therapeutic effect of MA on preventing bone loss in vivo was further confirmed in an ovariectomized mouse model. Therefore, considering its ability to inhibit RANKL-mediated osteoclastogenesis and the underlying mechanisms, MA might be a potential candidate for treating osteolytic bone diseases.
Subject(s)
Estrogens/metabolism , Osteogenesis/drug effects , Osteoporosis/drug therapy , RANK Ligand/metabolism , Triterpenes/pharmacology , Animals , Bone Resorption/drug therapy , Bone Resorption/metabolism , Cell Line , Centella , Female , Mice , Mice, Inbred C57BL , NF-kappa B/metabolism , NFATC Transcription Factors/metabolism , Osteoclasts/drug effects , Osteoclasts/metabolism , Osteoporosis/metabolism , Plant Extracts , Proto-Oncogene Proteins c-fos/metabolism , RAW 264.7 Cells , Signal Transduction/drug effects , Tartrate-Resistant Acid Phosphatase/metabolismABSTRACT
BACKGROUND: Random skin flaps are routinely placed during plastic and reconstructive surgery, but the distal areas often develop ischemia and necrosis. Baicalein, a major flavonoid extracted from the traditional Chinese herbal medicine huangqin, Scutellaria baicalensis Georgi, may improve flap viability. MATERIALS AND METHODS: Rats were randomly divided into baicalein and control groups and they underwent placement of modified McFarlane flaps after intraperitoneal administration of baicalein or vehicle. Flap survival and water content were measured 7 days later, as were angiogenesis, apoptosis, and oxidative stress in ischemic flaps. RESULTS: Baicalein promoted flap survival, reduced edema, increased mean vessel density, and enhanced vascular endothelial growth factor production at both the translational and transcriptional levels. Baicalein reduced caspase 3 cleavage, increased superoxidase dismutase and glutathione levels, and decreased the malondialdehyde level. CONCLUSION: Baicalein promoted flap viability by stimulating angiogenesis and inhibiting apoptosis and oxidation.
Subject(s)
Edema/drug therapy , Flavanones/pharmacology , Medicine, Chinese Traditional , Skin/drug effects , Surgical Flaps , Animals , Apoptosis/drug effects , Edema/metabolism , Edema/pathology , Male , Oxidative Stress/drug effects , Rats , Rats, Sprague-DawleyABSTRACT
Wound therapy remains a clinical challenge due to the poor vascularization during the healing process and the high demand to achieve functional and aesthetically satisfactory scars. Newly-formed blood vessels are necessary for wound healing since they can deliver nutrients and oxygen to the wound area. In this study, the role of leonurine (LN), a traditional Chinese medicine isolated from Herba leonuri, in promoting angiogenesis and its function in wound healing have been investigated. The results of co-culture with human umbilical vein endothelial cells (HUVECs) demonstrated that LN treatment (5-20 µM) could promote the proliferation and migration and enhance the ability of in vitro angiogenesis through up-regulating the mTOR/ERK signaling pathway. Furthermore, a full-thickness cutaneous wound model was used to investigate the healing effect of LN in vivo. Intragastric administration of 20 mg per kg per day LN stimulated the regeneration of more blood vessels at the wound sites, which confirmed the in vitro results of promoting angiogenesis. Due to fast vascularization, the collagen matrix deposition and remodeling processes were also accelerated in LN treated wounds, resulting in efficient wound healing. In summary, LN promoted angiogenesis of endothelial cells in vitro by activating the mTOR/ERK pathway, and could efficiently enhance the angiogenesis and collagen deposition of the regenerated tissue, together with facilitating the wound healing process in vivo. This study provides evidence for LN-stimulated angiogenesis and tissue regeneration in skin wounds, especially in ischemic wounds.
Subject(s)
Angiogenesis Inducing Agents/administration & dosage , Drugs, Chinese Herbal/administration & dosage , Extracellular Signal-Regulated MAP Kinases/metabolism , Gallic Acid/analogs & derivatives , Regeneration/drug effects , Skin/drug effects , TOR Serine-Threonine Kinases/metabolism , Wounds and Injuries/drug therapy , Animals , Cell Movement/drug effects , Disease Models, Animal , Extracellular Signal-Regulated MAP Kinases/genetics , Gallic Acid/administration & dosage , Human Umbilical Vein Endothelial Cells/drug effects , Human Umbilical Vein Endothelial Cells/metabolism , Humans , Neovascularization, Pathologic , Rats , Signal Transduction/drug effects , Skin/blood supply , Skin/metabolism , Skin/physiopathology , TOR Serine-Threonine Kinases/genetics , Wound Healing/drug effects , Wounds and Injuries/genetics , Wounds and Injuries/metabolism , Wounds and Injuries/physiopathologyABSTRACT
Attenuating oxidative stress-induced damage and promoting endothelial progenitor cell (EPC) differentiation are critical for ischaemic injuries. We suggested monotropein (Mtp), a bioactive constituent used in traditional Chinese medicine, can inhibit oxidative stress-induced mitochondrial dysfunction and stimulate bone marrow-derived EPC (BM-EPC) differentiation. Results showed Mtp significantly elevated migration and tube formation of BM-EPCs and prevented tert-butyl hydroperoxide (TBHP)-induced programmed cell death through apoptosis and autophagy by reducing intracellular reactive oxygen species release and restoring mitochondrial membrane potential, which may be mediated viamTOR/p70S6K/4EBP1 and AMPK phosphorylation. Moreover, Mtp accelerated wound healing in rats, as indicated by reduced healing times, decreased macrophage infiltration and increased blood vessel formation. In summary, Mtp promoted mobilization and differentiation of BM-EPCs and protected against apoptosis and autophagy by suppressing the AMPK/mTOR pathway, improving wound healing in vivo. This study revealed that Mtp is a potential therapeutic for endothelial injury-related wounds.
Subject(s)
Angiogenesis Inducing Agents/pharmacology , Antioxidants/pharmacology , Endothelial Progenitor Cells/drug effects , Iridoids/pharmacology , Surgical Wound/drug therapy , Wound Healing/drug effects , AMP-Activated Protein Kinases/genetics , AMP-Activated Protein Kinases/metabolism , Animals , Apoptosis/drug effects , Apoptosis/genetics , Autophagy/drug effects , Autophagy/genetics , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Bone Marrow Cells/metabolism , Carrier Proteins/genetics , Carrier Proteins/metabolism , Cell Movement/drug effects , Cell Proliferation/drug effects , Endothelial Progenitor Cells/cytology , Endothelial Progenitor Cells/metabolism , Gene Expression Regulation/drug effects , Intracellular Signaling Peptides and Proteins , Male , Neovascularization, Physiologic/drug effects , Oxidative Stress/drug effects , Phosphoproteins/genetics , Phosphoproteins/metabolism , Primary Cell Culture , Rats , Rats, Sprague-Dawley , Reactive Oxygen Species/antagonists & inhibitors , Reactive Oxygen Species/metabolism , Ribosomal Protein S6 Kinases, 70-kDa/genetics , Ribosomal Protein S6 Kinases, 70-kDa/metabolism , Signal Transduction , Surgical Wound/genetics , Surgical Wound/metabolism , Surgical Wound/pathology , TOR Serine-Threonine Kinases/genetics , TOR Serine-Threonine Kinases/metabolism , tert-Butylhydroperoxide/antagonists & inhibitors , tert-Butylhydroperoxide/pharmacologyABSTRACT
Wound therapy remains a clinical challenge due to the complexity of healing pathology and high demand of achieving functional and aesthetically satisfactory scars. Newly formed blood vessels are essential for tissue repair since they can support cells at the wound site with nutrition and oxygen. In this study, we investigated the effects of Asperosaponin VI (ASA VI) isolated from a traditional Chinese medicine, the root of Dipsacus asper Wall, in promoting angiogenesis, as well as its function in wound therapeutics. Treatment of human umbilical vein endothelial cells (HUVECs) with ASA VI (20-80 µg/mL) dose-dependently promoted the proliferation, migration and enhanced their angiogenic ability in vitro, which were associated with the up-regulated HIF-1α/VEGF signaling. Full-thickness cutaneous wound model rats were injected with ASA VI (20 mg·kg-1·d-1, iv) for 21 d. Administration of ASA VI significantly promoted the cutaneous wound healing, and more blood vessels were observed in the regenerated tissue. Due to rapid vascularization, the cellular proliferation status, granulation tissue formation, collagen matrix deposition and remodeling processes were all accelerated, resulting in efficient wound healing. In summary, ASA VI promotes angiogenesis of HUVECs in vitro via up-regulating the HIF-1α/VEGF pathway, and efficiently enhances the vascularization in regenerated tissue and facilitates wound healing in vivo. The results reveal that ASA VI is a potential therapeutic for vessel injury-related wounds.
Subject(s)
Hypoxia-Inducible Factor 1, alpha Subunit/physiology , Neovascularization, Physiologic/physiology , Saponins/pharmacology , Vascular Endothelial Growth Factor A/physiology , Wound Healing/drug effects , Animals , Cell Movement/physiology , Cell Proliferation/physiology , Cells, Cultured , Dose-Response Relationship, Drug , Drugs, Chinese Herbal/pharmacology , Humans , Rats , Signal Transduction/drug effects , Up-Regulation/drug effectsABSTRACT
Microglial activation leads to increased production of proinflammatory enzymes and cytokines, which is considered to play crucial role in neurodegenerative diseases, however there are only a few drugs that target microglia activation. Recent studies have indicated that the Traditional Chinese Medicine, salidroside (Sal), exerted anti-inflammatory effects. According to this evidence, our present study aims to explore the effect of the Sal (a phenylpropanoid glycoside compound which is isolated from rhodiola), on microglia activation in lipopolysaccharide (LPS)-stimulated BV-2 cells. Our results showed that Sal could significantly inhibit the excessive production of Nitric Oxide (NO) and Prostaglandin E2 (PGE2) in LPS-stimulated BV2 cells. Moreover, Sal treatment could suppress the mRNA and protein expressions of inflammatory enzymes, including Inducible Nitric Oxide Synthase (iNOS) and Cyclooxygenase-2 (COX-2). The mechanisms may be related to the inhibition of the activation of Nuclear Factor-kappaB (NF-[Formula: see text]B) and endoplasmic reticulum stress. Our study demonstrated that salidroside could inhibit lipopolysaccharide-induced microglia activation via the inhibition of the NF-[Formula: see text]B pathway and endoplasmic reticulum stress, which makes it a promising therapeutic agent for human neurodegenerative diseases.
Subject(s)
Anti-Inflammatory Agents , Endoplasmic Reticulum Stress/drug effects , Glucosides/pharmacology , Glucosides/therapeutic use , Microglia/pathology , NF-kappa B/metabolism , Neurodegenerative Diseases/drug therapy , Neurodegenerative Diseases/pathology , Phenols/pharmacology , Phenols/therapeutic use , Phytotherapy , Signal Transduction/drug effects , Animals , Cells, Cultured , Cyclooxygenase 2/metabolism , Dinoprostone/metabolism , Inflammation Mediators/metabolism , Mice , Neurodegenerative Diseases/metabolism , Nitric Oxide/metabolism , Nitric Oxide Synthase Type II/genetics , Nitric Oxide Synthase Type II/metabolism , RNA, Messenger/metabolismABSTRACT
A range of biological and molecular effects caused by nicotine are considered to effect bone metabolism. Vitamin C functions as a biological antioxidant. This study was to evaluate the in vitro effects of nicotine on human bone marrow stromal cells and whether Vitamin C supplementation show the antagonism action to high concentration nicotine. We used CCK-8, alkaline phosphatase (ALP) activity assay, Von Kossa staining, real-time polymerase chain reaction and Western Blot to evaluate the proliferation and osteogenic differentiation. The results indicated that the proliferation of BMSCs increased at the concentration of 50, 100 ng/ml, got inhibited at 1,000 ng/ml. When Vitamin C was added, the OD for proliferation increased. For ALP staining, we found that BMSCs treated with 50 and 100 ng/ml nicotine showed a higher activity compared with the control, and decreased at the 1,000 ng/ml. Bone morphogenetic protein-2 (BMP-2) expression and the calcium depositions decreased at 100 and 1,000 ng/ml nicotine, while the addition of Vitamin C reversed the down regulation. By real-time PCR, we detected that the mRNA expression of collagen type I (COL-I) and ALP were also increased in 50 and 100 ng/ml nicotine groups (P < 0.05), while reduced at 1,000 ng/ml (P < 0.05). When it came to osteocalcin (OCN), the changes were similar. Taken all together, it is found that nicotine has a two-phase effect on human BMSCs, showing that low level of nicotine could promote the proliferation and osteogenic differentiation while the high level display the opposite effect. Vitamin C could antagonize the inhibitory effect of higher concentration of nicotine partly.
Subject(s)
Antioxidants/pharmacology , Ascorbic Acid/pharmacology , Bone Marrow Cells/metabolism , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Ganglionic Stimulants/pharmacology , Nicotine/pharmacology , Osteogenesis/drug effects , Adult , Aged , Ascorbic Acid/antagonists & inhibitors , Bone Marrow Cells/cytology , Bone Morphogenetic Protein 2/biosynthesis , Cells, Cultured , Dose-Response Relationship, Drug , Drug Antagonism , Female , Ganglionic Stimulants/antagonists & inhibitors , Gene Expression Regulation/drug effects , Humans , Male , Middle Aged , Nicotine/antagonists & inhibitors , Stromal Cells/cytology , Stromal Cells/metabolismABSTRACT
Thymoquinone (TQ), the predominant bioactive constituent derived from the medicinal spice Nigella sativa (also known as black cumin), has been applied for medical purposes for more than 2,000 years. Recent studies reported that thymoquinone exhibited inhibitory effects on the cell proliferation of several cancer cell lines. This study was performed to investigate the antitumor and anti-angiogenic effects of thymoquinone on osteosarcoma in vitro and in vivo. Our results showed that thymoquinone induced a higher percentage of growth inhibition and apoptosis in the human osteosarcoma cell line SaOS-2 compared to that of control, and thymoquinone significantly blocked human umbilical vein endothelial cell (HUVEC) tube formation in a dose-dependent manner. To investigate the possible mechanisms involved in these events, we performed electrophoretic mobility shift assay (EMSA) and western blot analysis, and found that thymoquinone significantly downregulated NF-κB DNA-binding activity, XIAP, survivin and VEGF in SaOS-2 cells. Moreover, the expression of cleaved caspase-3 and Smac were upregulated in SaOS-2 cells after treatment with thymoquinone. In addition to these in vitro results, we also found that thymoquinone inhibits tumor angiogenesis and tumor growth through suppressing NF-κB and its regulated molecules. Collectively, our results demonstrate that thymoquinone effectively inhibits tumor growth and angiogenesis both in vitro and in vivo. Moreover, inhibition of NF-κB and downstream effector molecules is a possible underlying mechanism of the antitumor and anti-angiogenic activity of thymoquinone in osteosarcoma.
Subject(s)
Antineoplastic Agents/therapeutic use , Benzoquinones/therapeutic use , Bone Neoplasms/blood supply , Bone Neoplasms/drug therapy , NF-kappa B/metabolism , Osteosarcoma/blood supply , Osteosarcoma/drug therapy , Animals , Antigens, CD34/metabolism , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Benzoquinones/pharmacology , Bone Neoplasms/pathology , Caspase 3/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Human Umbilical Vein Endothelial Cells/drug effects , Humans , Inhibitor of Apoptosis Proteins/metabolism , Ki-67 Antigen/metabolism , Male , Mice , Mice, Inbred BALB C , Neovascularization, Pathologic/drug therapy , Oligopeptides/metabolism , Osteosarcoma/pathology , Signal Transduction/drug effects , Survivin , X-Linked Inhibitor of Apoptosis Protein/metabolismABSTRACT
OBJECTIVE: To study the effectiveness of retinoic acid on induction of osteoporotic rats treated by either alendronate or qianggu capsules and co-administration. METHODS: Sixty-five female SD rats were treated with retinoic acid 80 mg x kg(-1) x d(-1) by gastric lavage for 15 days. Then 5 rats were confirmed cases of osteoporosis and the remaining 60 were randomly divided into 4 groups 15 each: (1) control group with NS 8 ml x kg(-1) x w(-1); (2) alendronate group with alendronate 40 mg x kg(-1) x w(-1); (3) qianggu group with qianggu capsules 90 mg x kg(-1) x d(-1); (4) co-medicated group with alendronate 40 mg x kg(-1) x w(-1) and qianggu capsules 90 mg x kg(-1) x d(-1). Five rats in each group were sacrificed at week 2, 4 and 6 respectively to carry out the biomechanic tests, histopathologic examination and bony callus volume calculation. RESULTS: Biomechanical properties of femur changed significantly after the treatment by alendronate or qianggu capsules and co-medication as compared with that of NS after 4 weeks (P < 0.05); the bony callus were larger when treated by alendronate (P < 0.05) and smaller by qianggu capsules (P > 0.05); the bone trabecula formed and rebuilding were slower by alendronate and quicker by qianggu capsules. CONCLUSION: Alendronate or qianggu capsules and co-medication can improve biomechanical properties of femur by retinoic acid on induction of osteoporotic rats. Qianggu capsules can improve bone union.