ABSTRACT
Water-soluble porphyrins are considered promising drug candidates for photodynamic therapy (PDT). This study investigated the PDT activity of a new water-soluble, anionic porphyrin (1-Zn), which possesses four negative charges. The photodynamic anticancer activity of 1-Zn was investigated by the MTT assay, with mTHPC as a positive control. The cellular distribution was determined by fluorescence microscopy. Holographic and phase contrast images were recorded after 1-Zn treatment with a HoloMonitor™ M3 instrument. The inhibition of A549 cell growth achieved by inducing apoptosis was investigated by flow cytometry and fluorescence microscopy. DNA damage was investigated by the comet assay. The expression of apoptosis-related proteins was also measured by western blot assays. 1-Zn had better phototoxicity against A549 cells than HeLa and HepG2 cancer cells. Interestingly, 1-Zn was clearly located almost entirely in the cell cytoplasmic region/organelles. The late apoptotic population was less than 1.0% at baseline in the untreated and only light-treated cells and increased to 40.5% after 1-Zn treatment and irradiation (PĆ¢ĀĀÆ<Ć¢ĀĀÆ0.05). 1-Zn triggered significant ROS generation after irradiation, causing ΔΨm disruption (PĆ¢ĀĀÆ<Ć¢ĀĀÆ0.01) and DNA damage. 1-Zn induced A549 cell apoptosis via the mitochondrial apoptosis pathway. In addition, 1-Zn bound in the groove of DNA via an outside binding mode by pi-pi stacking and hydrogen bonding. 1-Zn exhibits good photonuclease activity and might serve as a potential photosensitizer (PS) for lung cancer cells.
Subject(s)
Photosensitizing Agents/chemical synthesis , Porphyrins/chemistry , Apoptosis/drug effects , Cell Cycle Checkpoints/drug effects , Cell Line, Tumor , DNA Damage/drug effects , Humans , Hydrogen Bonding , Light , Membrane Potential, Mitochondrial/drug effects , Neoplasms/drug therapy , Neoplasms/pathology , Photochemotherapy , Photosensitizing Agents/pharmacology , Photosensitizing Agents/therapeutic use , Porphyrins/pharmacology , Porphyrins/therapeutic use , Reactive Oxygen Species/metabolismABSTRACT
Corrole is a kind of new and promising photosensitizer (PS) in cancer photodynamic therapy (PDT). However, the protein molecular mechanism of PDT activity for corrole under light irradiation is still not clear. In this paper, water-soluble cationic sulfonated corrole (1) and its metal complexes (1-Fe, 1-Mn, and 1-Cu) were prepared, and the photodynamic anti-cancer activity against various tumor cells was investigated by MTT assay. The potential molecular mechanism of PDT activity was elucidated by fluorescence microscope, flow cytometry, molecular docking, and western blotting analysis. Besides, the potential PDT anti-tumor effect of 1 in vivo was assessed in human tumor xenografts in mice. Quantitative analysis revealed that 1's phototoxicity triggered a significant generation of reactive oxygen species, causing disruption of mitochondrial membrane potential. The results of western blotting (WB) assay shown in 1's phototoxicity could induce cell apoptosis via ROS-mediated mitochondrial caspase apoptosis pathway, in which SIRT1 protein degradation played a key role. PTD activity in vivo shown in 1 could significantly reduce the growth of A549 xenografted tumor, without obvious loss of mice body weight. We clearly found that cationic sulfonated corrole is a potential candidate of PS in vitro and in vivo. The phototoxicity of 1 could induce A549 cell apoptosis by inducing ROS production increase, further to activate the mitochondrial apoptosis pathway. We concluded that SIRT1 protein is a more appropriate target in this progress.
Subject(s)
Lung Neoplasms/drug therapy , Lung Neoplasms/pathology , Photochemotherapy , Porphyrins/therapeutic use , Sulfonic Acids/chemistry , Animals , Antineoplastic Agents/pharmacology , Antineoplastic Agents/therapeutic use , Apoptosis/drug effects , Apoptosis/radiation effects , Caspases/metabolism , Cations , Cell Cycle/drug effects , Cell Cycle/radiation effects , Cell Line, Tumor , Humans , Membrane Potential, Mitochondrial/drug effects , Membrane Potential, Mitochondrial/radiation effects , Mice, Inbred BALB C , Mice, Nude , Mitochondria/metabolism , Molecular Docking Simulation , Photosensitizing Agents/pharmacology , Porphyrins/chemistry , Porphyrins/pharmacology , Reactive Oxygen Species/metabolism , Signal Transduction/drug effects , Sirtuin 1/metabolism , Xenograft Model Antitumor AssaysABSTRACT
Guaiane-type sesquiterpenoids are most prevalent in the genus Cinnamomum. Hence this study investigates the structures, anti-nociceptive and IL-6 targeted anti-inflammatory potential of three novels C-14 guaiane-type sesquiterpenoids and two new monoterpenoids, isolated from Cinnamomum migao. The structures were precisely confirmed and characterized through the modern chromatographic and spectroscopic techniques of HRESIMS, 1D NMR, 2D NMR, experimental circular dichroism (ECD), and calculated (ECD). Novel sesquiterpenoids 1 and 2 exhibited significant anti-inflammatory activities against the NO production and pro-inflammatory cytokines. Their IC50 values were determined as 9.52 and 13.42 ĀµΜ against IL-6 mRNA, respectively. Similarly, subcutaneous injection of n-BuT and EA extracts showed a dose-dependent suppression of formalin-induced tonic biting/licking responses during the tonic antinociceptive phase. Furthermore, absorption, distribution, metabolism, excretion, and toxicity (ADMET) analysis of guaiane-type sesquiterpenoids 1 and 2 displayed that both compounds have a high level of GIT absorption, with a high zone of safety for cardiac and hepatotoxicity and no inhibition of cytochromes. In addition, molecular docking and simulation studies strengthen the anti-inflammatory potential of sesquiterpene 2 which showed a good binding affinity with IL-6 protein. Overall the inclusive results showed that the extracts and newly isolated guaiane-type sesquiterpenoids from C. migao will provide new evidence for the traditional use of this species to treat inflammation and nociception.
Subject(s)
Interleukin-6 , Sesquiterpenes , Molecular Docking Simulation , Molecular Structure , Anti-Inflammatory Agents/pharmacology , Sesquiterpenes, Guaiane/pharmacology , Plant Extracts , Sesquiterpenes/chemistryABSTRACT
Osteomyelitis is an acute or chronic inflammatory bone disease with a high disability rate. As an anti-inflammatory factor, peroxisome proliferator activated receptor-ĆĀ³ (PPAR-ĆĀ³) is not only implicated in a variety of inflammatory responsesĀ but also regulates osteoblast differentiation and bone mass. However, the role of PPAR-ĆĀ³ in osteomyelitis is not fully understood. In the present study, we demonstrated that PPAR-ĆĀ³ showed a lower expression level in infected bone tissue of osteomyelitis patients as compared with uninfected bone tissue from nonosteomyelitis patients with fracture of the hip. We applied lipopolysaccharides (LPSs) in MC3T3-E1 osteoblast precursor cell line as an in vitro model for osteomyelitis. LPS treatment increased osteomyelitis-associated inflammatory cytokines such as interleukin-6Ā (IL-6) and tumor necrosis factor-α (TNF-α), whereas PPAR-ĆĀ³ levels and cell viability in MC3T3-E1 cells were suppressed. PPAR-ĆĀ³ antagonist GW9662 further enhanced IL-6 and TNF-α levels, and decreased cell viability in the presence of LPS treatment. In contrast, PPAR-ĆĀ³ agonist pioglitazone antagonized the effect of LPS treatment in MC3T3-E1 cells. These findings suggest that PPAR-ĆĀ³ downregulation is associated with the inflammation and progression of osteomyelitis, and PPAR-ĆĀ³ agonist could serve as a therapeutic strategy to attenuate inflammatory responses. This study provides novel insights into the physiopathogenesis of osteomyelitisĀ and future study is required to validate the findings in animal model and uncover the molecular mechanism of PPAR-ĆĀ³-dependent anti-inflammation in osteoblasts.
Subject(s)
Osteomyelitis , Thiazolidinediones , Animals , Anti-Inflammatory Agents/pharmacology , Cytokines , Hypoglycemic Agents , Interleukin-6/metabolism , Lipopolysaccharides/pharmacology , Osteoblasts/metabolism , Osteomyelitis/drug therapy , PPAR gamma/metabolism , Pioglitazone/pharmacology , Pioglitazone/therapeutic use , Thiazolidinediones/pharmacology , Thiazolidinediones/therapeutic use , Tumor Necrosis Factor-alpha/metabolismABSTRACT
OBJECTIVE: To compare clinical effects of artificial bone and autogenous bone in internal fixation of complex calcaneal fracture with profitated plate. METHODS: From April 2015 to April 2019, 60 patients with complex calcaneal fractures were treated with open reduction and heteromorphic plate internal fixation, and were divided into experiment group and control group by implant bone substitutes, and 30 patients in each group. In experiment group, there were 21 males and 9 females aged from 18 to 71 years old with an average of (36.85Ā±7.42) years old;19 patients were classified to type Ć¢Ā Ā¢ and 11 patients were type Ć¢Ā Ā£ according to Sanders classification;implanted with artificial bone. While in control group, there were 23 males and 7 females aged from 20 to69 years old with an average of (37.26Ā±7.38) years old;18 patients were classified to type Ć¢Ā Ā¢ and 12 patients were type Ć¢Ā Ā£ according to Sanders classification; implanted with autogenous bone. Operation time, intraoperative blood loss, drying time of incision, fracture healing time and complications were compared between two groups, changes of preoperative and postoperative Bƶhler angle and Gissane angle were also compared, and Maryland scoring was applied to evaluate recovery of affected foot. RESULTS: Both of two groups were followed up from 3 to 15 months with an average of (10.15Ā±2.67) months. Operation time and intraoperative blood loss in experiment group were (89.32Ā±12.43) min, (101.64Ā±5.13) ml, respectively;while in control group were (112.45Ā±13.82) min, (119.01Ā±5.26) ml, respectively;and there were statistical difference between two groups (P<0.05). There were no differences in drying time of incision, fracture healing time and complications between two groups(P>0.05). Preoperative and postoperative Bƶhler angle at 3 months in experiment group were (14.19Ā± 2.47)Ā°, (34.52Ā±4.41)Ā°;while in control group were (14.08Ā±2.35)Ā°, (33.67Ā±4.36)Ā°;preoperative and postoperative Gissane angel at 3 months in experiment group were (90.16Ā±6.43)Ā°, (131.45Ā±9.83)Ā°;while in control group were (90.11Ā±6.37)Ā°, (130.87Ā±9.24)Ā°;there were statistical significance in Bƶhler angle and Gissane angel compared with 3 months after operation and before operation between two groups (P<0.05);while no difference at 3 months after operation between two groups (P> 0.05). There were no statistical meaning in Maryland items, total score and good rate between two groups at the final following up (P>0.05). CONCLUSION: Artificial bone and autogenous bone in internal fixation of complex calcaneal fracture with irregular plate have similar function in promoting fracture healing, drying time of incision, fracture healing time and complications, while artificial bone has better effects in reducing intraoperative blood loss, shorten operation time.
Subject(s)
Calcaneus , Fractures, Bone , Adolescent , Adult , Aged , Bone Plates , Calcaneus/surgery , Case-Control Studies , Female , Fracture Fixation, Internal , Fractures, Bone/surgery , Humans , Male , Middle Aged , Treatment Outcome , Young AdultABSTRACT
The development of novel, efficient and nontoxic photosensitizers (PSs) is a challenging task for photodynamic therapy (PDT). In our previous study, corrole had been demonstrated to be a promising PS in PDT for cancer cells. In this paper, a novel electron-deficient flat phosphorus tris(ethoxycarbonyl) corrole (1-P) was synthesized and characterized. In vitro photodynamic activities and toxicity of 1-P in HepG2 xenograft tumours was evaluated by standard assay. The results shown 1-P displayed a potential efficient and low-toxic PS, which suggesting this kind of corrole is a powerful and promising antitumor PS for PDT. In addition, the potential anti-tumour mechanism study of 1-P was also investigated by the apoptosis antibody array, immunohistochemical and western blotting assay (WB) experiments, we found that the PDT activity of 1-P can degrade SIRT1 (an important deacetylase), and activate the Fas signal pathway to inhibit the growth of liver cancer cells.
Subject(s)
Photochemotherapy/methods , Photosensitizing Agents/pharmacology , Animals , Apoptosis/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Hep G2 Cells , Humans , Liver Neoplasms/drug therapy , Liver Neoplasms/pathology , Mice , Phosphorus/pharmacology , Photosensitizing Agents/chemistry , Photosensitizing Agents/toxicity , Porphyrins/pharmacology , Sirtuin 1/drug effects , Xenograft Model Antitumor Assays , fas Receptor/metabolismABSTRACT
The present study investigated the effect of microRNAĀ (miR)Ā15aĀ3p on the proliferation, migration and apoptosis of lens epithelial cells and its potential mechanism, in order to further elucidate the pathogenesis of ageĀrelated cataractsĀ (ARCs). The HLEĀB3 human lens epithelial cell line was transfected with miRĀ15aĀ3p mimic. Expression of the miRĀ15aĀ3p mimic was measured by fluorescenceĀbased reverse transcriptionĀquantitative polymerase chain reaction analysis. Cell proliferation, apoptosis, invasion and migration were investigated using MTT and plate clone formation assays, terminal deoxynucleotidyl transferase dUTP nick end labeling and flow cytometry, and a wound healing assay and Transwell assay, respectively. The protein expression levels of BĀcell lymphomaĀ 2Ā (BCL2) and myeloid cell leukemia sequenceĀ 1Ā (MCL1) were also compared between transfected and wildĀtype HLEĀB3 cells by western blot analysis. The results showed that transfection with the miRĀ15aĀ3p mimic significantly suppressed the proliferation of HLEĀB3 cells, induced cell apoptosis and increased the proportion of early apoptotic cells. The migration of HLEĀB3 cells was significantly inhibited following transfection with miRĀ15aĀ3p mimic (P<0.01), whereas cell invasion was unaffected (P>0.05). In addition, reduced protein levels of BCL2 and MCL1 were observed in the miRĀ15aĀ3p mimicĀtransfected HLEĀB3 cells (P<0.01). In conclusion, miRĀ15aĀ3p may suppress cell proliferation and migration, and induce cell apoptosis in lens epithelial cells through inhibiting the expression of BCL2 and MCL1, which contributes to the onset of ARCs.
Subject(s)
Apoptosis/genetics , Gene Expression Regulation, Neoplastic , Lens, Crystalline/metabolism , MicroRNAs/genetics , Myeloid Cell Leukemia Sequence 1 Protein/genetics , Proto-Oncogene Proteins c-bcl-2/genetics , Aged , Antagomirs/genetics , Antagomirs/metabolism , Cataract/genetics , Cataract/metabolism , Cataract/pathology , Cell Line, Transformed , Cell Movement , Cell Proliferation , Epithelial Cells/metabolism , Epithelial Cells/pathology , Humans , Lens, Crystalline/pathology , MicroRNAs/agonists , MicroRNAs/antagonists & inhibitors , MicroRNAs/metabolism , Models, Biological , Myeloid Cell Leukemia Sequence 1 Protein/metabolism , Oligoribonucleotides/genetics , Oligoribonucleotides/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Signal Transduction , TransfectionABSTRACT
OBJECTIVE: To evaluate the middle-long-term clinical effects of unilateral and bilateral percutaneous kyphoplasty(PKP) for vertebral fragility fracture in the elderly. METHODS: The clinical data in elderly patients with vertebral fragility fracture treated by unilateral and bilateral PKP between January 2008 and January 2010 was retrospective analyzed. According to a unified criteria to divided into two groups for 104 patients(44 males and 60 females), of them, 50 cases were divided in unilateral group using unilateral pedicle surgical approach for PKP and 54 cases were in bilateral group using bilateral pedicle approach. VAS score, Cobb angle, and the height of anterior and posterior vertebral body were respectively analyzed peroperatively and at 3 d, 3 months, 1 year, 3 years postoperatively and final follow-up. Clinical effects and safety were assessed in two groups. RESULTS: All the operations were successful. Operative time and bone cement injection volume in unilateral group were less than those of bilateral group(P<0.05). Postoperative VAS scores, Cobb angle, and the height of anterior and posterior vertebral body were obviously improved in two groups(P<0.05), and there was no significant difference between two groups(P>0.05). Bone cement leakage occurred in 12 cases(11.5%), recurrent fracture of vertebral body occurred in 5 cases(4.8%), cerebrospinal leak occurred in 2 cases(1.9%), and nerve root irritation occurred in 3 cases(2.9%). The above complications were transient and released after symptomatic treament. CONCLUSIONS: Middle-long-term clinical effects of unilateral and bilateral percutaneous kyphoplasty for vertebral fragility fracture are safe and satisfactory, and may be extended as a minimally invasive method. Unilateral approach for PKP has advantages of short operation time, relative less trauma, thus is a more ideal method.
Subject(s)
Bone Cements/therapeutic use , Kyphoplasty/methods , Osteoporotic Fractures/surgery , Spinal Fractures/surgery , Aged , Bone Cements/adverse effects , Female , Humans , Male , Operative Time , Pain Measurement/methods , Retrospective Studies , Treatment OutcomeABSTRACT
OBJECTIVE: To introduce and report the preliminary results of 7 old patients with fractures and dislocations of the elbow. METHODS: From July 2011 to August 2015, 7 old patients suffered from fractures and dislocations of the elbow(5 of which were terrible triad). One patient had type Iradial head fracture, 3 type IIand 1 type III according to the Mason classification, and 1 type I, 5 type IIand 1 type III according to the Regan-Morrey classification. All the 7 patients received operation and then were treated with external fixation. Fractures of the radial head were fixed with Herbert screws or locking plates and screws. Fractures of ulnar coronoid were reduced and fixed with lag screws or K-wires or PDS sutures or locking screws according to the types. The lateral and medial collateral ligaments were also repaired. Plaster external fixation was applied for 3 weeks after operation, in the position with elbow flexion in 90 degrees and forearm rotation in neutral. External fixation braces were used for each patient after the plasters were removed, and at the same time rehabilitation programs were carried out. RESULTS: All the 7 patients were followed up, and the during ranged from 13 to 48 months(averaged, 20 months), with healed fractures, stable elbow and no pain movement. The functional outcome was excellent in 3 patients, good in 3 and fair in 1 according to the Mayo Elbow Performance Score(MEPS). CONCLUSIONS: It is not easy to get stable fixation for fractures and dislocations of the elbow in old patients with osteoporosis and low density of bone, but the operation can achieve satisfied clinical outcomes after external fixation.
Subject(s)
Elbow Injuries , Joint Dislocations/surgery , Osteoporotic Fractures/surgery , Radius Fractures/surgery , Radius/injuries , Aged , Humans , Range of Motion, Articular , Treatment OutcomeABSTRACT
Corroles have been previously demonstrated to be a new and promising photosensitizer (PS) in photodynamic therapy (PDT) for cancer cells. This research reported the preparation of an electron-deficient flat gallium(iii) tris(ethoxycarbonyl)corrole (1-Ga). Its in vitro PDT activities towards different cancer cell lines were examined, and the best PDT activity was observed in A549 cell lines. 1-Ga could penetrate the cell membrane rapidly and exhibit remarkable photo-cytotoxicity. 1-Ga can induce A549 cell apoptosis via ROS-mediated mitochondrial pathways, in which p38 and sirt-1 proteins play a key role. A PTD test in vivo showed that 1-Ga can significantly reduce the growth of A549 xenografted tumor cells without an obvious loss of mice weight upon PDT treatment.
ABSTRACT
PURPOSE: To investigate the effect of topical administration of peroxiredoxin-6 (PRDX6) on ultraviolet-induced corneal injury. METHODS: Corneal transparency and neovascularization were observed with a slit-lamp microscope and hematoxylin and eosin staining. The oxidative damage was determined with a commercial malondialdehyde (MDA) kit. The expressions of PRDX6, polymorphonuclear neutrophil (PMN), vascular endothelial growth factor (VEGF), and pigment epithelium-derived factor (PEDF) were determined by immunohistochemistry and Western blot. The expressions of genes related with antioxidant defense systems and cell apoptosis were detected by RT-PCR. RESULTS: The irradiated corneas appeared opaque and had high levels of MDA. Peripheral neovascularization and neutrophils appeared in the control and buffer-treated groups (with no treatment or PRDX6 diluent, respectively), whereas they were significantly suppressed in the PRDX6-treated group. The MDA content of the corneas in the PRDX6-treated group was significantly lower than that of the control and buffer-treated groups (P < 0.05). In the PRDX6-treated group the immunoreactivity of VEGF was lower, and that of PEDF was higher, than that in the control and buffer-treated groups. In addition, there were expression correlations between PRDX6 and PMN, VEGF, PEDF. The expressions of genes related with antioxidant defense systems and cell apoptosis were significant different between buffer- and PRDX6-treated groups (P < 0.05). CONCLUSIONS: The topically administered PRDX6 maintained the homeostasis of corneal cells, reduced inflammation, and suppressed neovascularization and apoptosis under ultraviolet irradiation.