ObjectiveAfter using hyaluronic acid (HA) to modify curcumin (CUR), the effects of calcium phosphate cement (CPC) combined with HA/CUR on the proliferation and osteogenesis of osteoblasts were investigated.MethodsFirst, HA and CUR were esterified and covalently combined to prepare HA/CUR, and the characteristics were observed and the infrared spectrum was tested. Then, HA, CUR, and HA/CUR were mixed with CPC according to 5% (W/W) to prepare HA-CPC, CUR-CPC, and HA/CUR-CPC, respectively. Setting time detection, scanning electron microscope observation, injectable performance test, and compression strength test were conducted; and the CPC was used as a control. Osteoblasts were isolated and cultured from the skull of newborn Sprague Dawley rats, and the 2nd generation cells were cultured with the 4 types of bone cement, respectively. The effects of HA/CUR-CPC on the proliferation and osteogenesis of osteoblasts were estimated by the scanning electron microscopy observation, live/dead cell fluorescence staining, cell counting, osteopontin (OPN) immunofluorescence staining, alkaline phosphatase (ALP) staining,and alizarin red staining.ResultsInfrared spectroscopy test showed that HA and CUR successfully covalently combined. The HA/CUR-CPC group had no significant difference in initial setting time, final setting time, injectable rate, and compressive strength when compared with the other 3 groups (P>0.05); scanning electron microscope observation showed that HA/CUR was scattered on CPC surface. After co-culture of bone cement and osteoblasts, scanning electron microscopy observation showed that the osteoblasts, which had normal morphology and the growth characteristics of osteoblasts, clustered and adhered to HA/CUR-CPC. There was no significant difference in cell survival rate between HA/CUR-CPC group and other groups (P>0.05), and the number of cells significantly increased (P<0.05); the degrees of OPN immunofluorescence staining, ALP staining, and alizarin red staining were stronger than other groups.ConclusionHA/CUR-CPC has good biocompatibility and mechanical properties, which can promote the proliferation and osteogenesis of osteoblasts.
Objective To observe the therapeutic effect of thermosensitive hydrogel containing curcumin-vitamin E (VE) complex (hereinafter referred to as “curcumin-VE hydrogel”) on radiation-induced oral mucositis in mice. Methods Curcumin-VE hydrogel was prepared using the synthesized curcumin-VE complex as the carrier and poloxam as the substrate. The structure of curcumin-VE complex was characterized by Fourier transform infrared spectrometer, the microstructure of curcumin-VE hydrogel was determined by scanning electron microscope, and the gelation temperature was determined by rheometer, gel swelling and degradation were tested and gel adhesion was determined using a universal testing machine. Thirty healthy male BALB/C mice with specific pathogen free grade were randomly divided into three groups, with ten mice in each group. The radiation group and radiation+hydrogel group were modeled by a single high dose of radiation (25 Gy), while the control group had anesthesia but no radiation. The control group and radiation group were given daily feed and water 7 days after radiation. In addition to daily feed and water, the radiation+hydrogel group was given curcumin-VE hydrogel twice a day. The mice were sacreficed on the 8th day after radiation. The weight changes of each group were recorded after radiation. The ulceration area of tongue was measured by toluidine blue. The tongue of mouse were pathologically observed. The activities of superoxide dismutase, catalase (CAT), and glutathione peroxidase and the level of malondialdehyde in tongue tissue were determined. The levels of tumor necrosis factor α (TNF-α), interleukin (IL)-1β and IL-6 in tongue tissue were determined by enzyme linked immunosorbent assay. The distribution and positive expression of phosphorylated histone H2AX (γ-H2AX) and nuclear factor-erythroid 2-related factor 2 were determined by immunohistochemistry. Results Curcumin-VE hydrogel had a porous network structure and the gelation temperature was 30℃, the swelling rate was close to 300%, the gel degradation rate was up to 95% after 48 h, and the adhesion strength was 12.748 kPa. Compared with the radiation group, the weight of mice in the radiation+hydrogel group increased (P<0.05), the ulcer area decreased (P<0.05); the activity of CAT increased (P<0.05); the levels of TNF-α, IL-1β and IL-6 decreased (P<0.05); the expression of γ-H2AX was down-regulated (P<0.05). Conclusion Curcumin-VE hydrogel can delay or weaken the process of radiation-induced oral mucositis by reducing the DNA damage caused by radiation, inhibiting the production of reactive oxygen species, and effectively reducing the level of inflammation in tongue tissue.
Objective To investigate the proliferation inhibitory effect and to explore the molecular mechanism of curcumin on pulmonary fibroblasts. Methods Fibroblasts derived from lung tissue of patients with idiopathic pulmonary fibrosis ( IPF) was cultured in vitro and incubated with curcumin at different concentrations for different time. Fibroblasts were randomized into 5 groups, ie. a control group and 4 curcumin groups ( intervened by 5, 10, 20, 40 μmol / L curcumin, respectively) . MTT assay was used to determine the inhibitory rate of curcumin on the proliferation of pulmonary fibroblasts. Apoptosis and the Caspase-3 expression of pulmonary fibroblasts were identified by flow cytometry ( FCM) . Variables were compared with One-Way ANOVA. The correlations between variables were analyzed using Pearson’scorrelation coefficient. Results Curcumin inhibited pulmonary fibroblasts proliferation in a dose-dependent and time-dependent manner( r =0. 886, r = 0. 832, respectively, all P lt; 0. 01) . Apoptosis rate of pulmonary fibroblasts in 4 curcumin groups was ( 29. 58 ±2. 13) % , ( 64. 36 ±3. 92) %, ( 72. 98 ±4. 42) % , ( 83. 14 ±2. 51) % , respectively, which was significantly higher than that in the control group[ ( 3. 84 ±1. 88) % , P lt;0. 01] . The positive expression rate of apoptosis-regulating protein caspase-3 was ( 26. 24 ±3. 64) % ,( 44. 87 ±5. 31) % , ( 57. 44 ±4. 23) % , ( 73. 65 ±5. 01) % , respectively, which was significantly higher than that of the control group[ ( 4. 02 ±0. 62) % , P lt; 0. 01] . Conclusions In vitro, curcumin can significantly inhibit proliferation and induce apoptosis of pulmonary fibroblasts of patients with IPF. The mechanism maybe associated with up-regulating expression of Caspase-3.
Objective To study the inhibitory effects of curcumin on bleomycin-induced pulmonary fibrosis in rats at the fibrosing stage and explore its possible mechanism.Methods 96 male SD rats were randomly divided into a normal control group，a fibrosis model group，a fibrosis model treated with prednisone group and a fibrosis model treated with curcumin group.Pulmonary fibrosis were induced by instilled bleomycin through tracheal.From day 15 after bleomycin administration，the curcumin group and prednisone group were given curcumin（300 mg/kg） or prednisone（5 mg/kg） per day by intragastric administration，respectively.The normal control group and fibrosis model group were given 1% sodium carboxymethyl cellulose（10 mL/kg） as control.Six rats of each group were randomly sacrificed on day 21，28，42 and 56 after bleomycin administration，respectively.The histological changes of the lung were evaluated by HE and Masson’s trichrome staining.Lung expressions of transforming growth factor-β1（TGF-β1） and hydroxyproline were assessed by immuno-histochemistry and digestion method，respectively.Results Pulmonary fibrosis and hydroxyproline level in the curcumin group were significantly reduced as compared with those in the model group on day 42 and 56.The expession of TGF-β1 in the curcumin group was significantly lower than that in the model group on day 28，42 and 56，and was not significantly different from the normal group on day 56.Conclusion Curcumin could alleviate bleomycin-induced pulmonary fibrosis in rats at the fibrosing stage by inhibiting the expressions of TGF-β1.
ObjectiveTo observe the effects of Curcumin on the cellular apoptosis of rat retinal vascular endothelial cells (RRVEC) induced by high glucose.MethodsGeneration 4 cultured RRVEC were used in this experiment, and identified with anti-vWF factor antibody by immunochemistry and immunofluorescence. The RRVEC were divided into control group (5.5 mmol/L glucose), high glucose group (30 mmol/L glucose), and treatment group (30 mmol/L glucose+30 μmol/L Curcumin), respectively. Flow cytometry was used to measure the cellular reactive oxygen species (ROS) level and apoptosis. The expression intensity and location of nuclear factor (NF)-κB p65 in the cells of the three groups were detected by immunochemistory. The expression of Bcl-2 and Bax protein was detected by Western blot test.ResultsImmunostaining showed that RRVEC were positive for vWF factor. The flow cytometry showed that the cellular ROS level in treatment group was higher than that in the control group (t=8.677, P=0.000), but less than that in the high glucose group (t=40.957, P=0.000). Compared with the high glucose group, the cellular ROS level in the treatment group was decreased significantly (t=6.568, P=0.000). The cellular apoptosis were significantly different among the three groups (F=325.137, P=0.000). Compared with the high glucose group, the cellular apoptosis in the treatment group was decreased significantly (t=12.818, P=0.000). Immunochemistry showed that NF-κB p65 was expressed strongly in the cellular nuclei and cytoplasm in the high glucose group than that in the control group and the treatment group with the significant differences (t=8.322, P=0.000). Western blot results demonstrated that compared with the control group, the expression of Bcl-2 of RRVEC and Bcl-2/Bax ratio decreased (t=4.362, 6.449; P=0.005, 0.001) and Bax increased (t=3.813, P=0.009）in the high glucose group, with statistically significant differences. Compared with the high glucose group, the expression of NF-κB and Bax decreased (t=2.577, 3.059; P=0.042, 0.022) and Bcl-2/Bax ratio increased significantly (t=3.831, P=0.009) in the treatment group.ConclusionCurcumin could suppress the cellular apoptosis of RRVEC induced by high glucose. The mechanism of Curcumin protecting RRVEC may be via regulating NF-κB signal pathway.
ObjectiveTo investigate the effects of curcumin on expression of iNOS mRNA, study the protective and therapeutic effects on rats with severe acute panceratitis associated renal injury, and explore the possible mechanisms. MethodsA total of 24 rats were randomly divided into control group (n=8), severe acute panceratitis with renal injury group (injury group, n=8), and curcumin-treated group (treated group, n=8).The rats of control group and injury group were given 1.5 mL saline through intragastric administration at 3 h before operation, while the rats of treated group were fed with same amount of curcumin diluent at 3 h before operation.The rats of injury group and treated group with pancreatic head clamping method to establish the model of severe acute pancreatitis.At 12 h after modeling, rats in control group and injury group were perfused with 1.5 mL saline, and rats in treated group were intragastrically administrated to the same volume of curcumin diluent.All the rats were sacrificed at 18 h after modeling.The levels of serum amylase, creatinine, and blood urea nitrogen were detected and pathological chang of pancreas and the left kidney were observed under light microscope.The renal cell apoptosis were analyzed using TUNEL staining in three groups.The serum levels of interleukin-1β(IL-1β), interleukin-6 (IL-6), and interleukin-10 (IL-10) in three groups were detected by enzyme-linked immunosorbert assay (ELISA).The expressions of inducible nitric oxide synthase (iNOS) mRNA in the right kidney were detected with realtime polymerase chain reaction (RT-PCR) in three groups. ResultsCompared with the control group, the levels of serum amylase, creatinine, blood urea nitrogen, IL-1β, IL-6, and IL-10, the cell apoptosis and the expressions of iNOS mRNA in injury group and treated group were significantly increased (P < 0.05).In the treated group, the level of IL-10 was higher than the injury group (P < 0.05), and the difference of other indexes were lower than the injury group (P < 0.05). ConclusionsCurcumin has a better protective effect on severe acute pancreatitis associated renal injury.It may be through inhibited the expressions of IL-1βand IL-6, stimulated the expression of IL-10, and down-regulation of iNOS mRNA expression to reduce the generation of oxygen free radicals and NO damage to cell and reducing the cell apoptosis and necrosis of kidney.
Objective To investigate the effect of curcumin on calcitionin gene related peptide (CGRP) expression after spinal cord injury (SCI) in rats. Methods A total of 200 rats, weighing 250-300 g, were randomly divided into 4 groups (n=50): sham-operation group, normal saline (NS) group, low-dose curcumin group (30 mg/kg), and high-dose curcumin group (100 mg/kg). In sham-operation group, only vertebral lamina excision was performed without SCI; the SCI model was established in the other 3 groups. At immediate after modeling, 30 mg/kg and 100 mg/kg curcumin were injected intraperitoneally in 2 curcumin groups, equivalent NS was given in NS group (30 mg/kg), but no treatment in sham-operation group. At 1, 3, 7, 14, and 21 days after operation, the motor neural function was evaluated by the inclined plane test and Basso-Beattie-Bresnahan (BBB) scores; immunohistochemical staining and Western blot assay were used to observe CGRP expression. Results BBB score and inclined plane test score of NS group, low-dose curcumin group, and high-dose curcumin group were significantly lower than those of sham-operation group at each time point (P lt; 0.05). BBB score of low-dose curcumin group and high-dose curcumin group was significantly higher than that of NS group at 3, 7, 14, and 21 days after SCI (P lt; 0.05), and the score of high-dose group was significantly higher than that of low-dose curcumin group at 7, 14, and 21 days after SCI (P lt; 0.05). Inclined plane test score of low-dose curcumin group and high-dose curcumin group was significantly higher than that of NS group at 7, 14, and 21 days after SCI (P lt; 0.05), and the score of high-dose curcumin group was significantly higher than that of low-dose curcumin group at 7, 14, and 21 days after SCI (P lt; 0.05). Immunohistochemical staining results showed that the CGRP positive cells of sham-operation group was significantly more than those of the other 3 groups, and the CGRP positive cells of high-dose curcumin group were significantly more than those of low-dose curcumin group at each time point (P lt; 0.05); the CGRP positive cells of low- and high-dose curcumin groups were significantly more than those of NS group at 3, 7, 14, and 21 days after SCI (P lt; 0.05). Western blot assay results showed that the CGRP protein expressed at each time point after SCI in sham-operation group; the CGRP protein expression gradually decrease with time passing in NS group; but the CGRP protein expression gradually increased with time passing in low- and high-dose curcumin groups, and reached the peak at 14 days, then maintained a high level. Conclusion After SCI in rats, 30 mg/kg curcumin can improve rats’ motor function, and 100 mg/kg curcumin effect is more obvious, especially in promoting the expression of CGRP. That may be the mechanism of protection of the nervous system.
Objective To investigate the effects of curcumin on oxidative stress in the co-culture system including human fetal lung fibroblasts and A549 cells, and discuss the potential and protective mechanism of the prophylactic effect of curcumin on pulmonary fibrosis. Methods The human fetal lung fibroblasts co-cultured with A549 cells were divided into five groups. The cells in the control group were cultured in DMEM without TGF-β1 or curcumin. The cells in the TGF-β1 group were cultured in DMEM containing 5 ng/mL TGF-β1 . In three TGF-β1 + cucurmin treatment groups, the cells were cultured in DMEM containing 5 ng/mL TGF-β1 and three different concentration of curcumin( 5, 10, 20 μmol /L, respectively) . ELISA was used to analyze the content of TNF-α. Serum level of MDA and SOD were tested by spectrophotometric analysis. Intracellular ROS production was detected by flow cytometry. NF-κB was measured by western blot. Results The serum MDA, intracellular ROS, the content of TNF-αand NF-κB protein expression in the TGF-β1 group were significantly increased while the activity of SOD was significantly decreased( P lt; 0. 01) , suggesting that the oxidative level of human fetal lung fibroblasts was obviously increased after TGF-β1 stimulation. After intervening by different concentration of curcumin, the serum MDA, intracellular ROS, content of TNF-αand NF-κB were significantly decreased while the activity of SOD was obviously increased( P lt;0.01) . Conclusion Low concentration of curcumin can reduce the oxidative level of human fetal lung fibroblasts co-cultured with A549 after TGF-β1 stimulation, and significantly increase the level of SOD, implying that curcumin may intervene pulmonary fibrosis by reduce oxidative level.
ObjectiveTo investigate the effect of Curcumin combined with Rhodiola on rats with severe acute pancreatitis (SAP) associated renal injury and explore the possible mechanisms. MethodsA total of 24 rats were randomly divided into SAP with renal injury group (SAP group, n=8), Curcumin group (n=8), Curcumin combined with Rhodiola group (n=8).The SAP group was given 1.5 mL saline through intragastric administration before operation while the Curcumin group was fed with same amount of Curcumin diluent.The Curcumin combined with Rhodiola group was given 1.5 mL Curcumin diluent through intragastric administration and 6 g/kg Rhodiola diluent through intraperitoneal injection before operation.The pancreas and pancreatic tail-segment was dissociated and the head of pancreas were occluded in rats to make the model, blood vessel forceps was loosed after three hours.All the rats were sacrificed at 18 h after modeling.The levels of serum amylase, creatinine, blood urea nitrogen were detected and pathological changes of pancreas and the left kidney were observed under the light microscope.The cell apoptosis was analyzed using TUNEL staining.The serum levels of interleukin (IL)-1β, IL-6, and IL-10 among the three groups were detected by enzyme-linked immunosorbent assay.The expression of inducible nitric oxide synthase (iNOS) mRNA in the right kidney was detected by real-time polymerase chain reaction.The superoxide dismutase (SOD) activity of the renal tissue was determined by hydroxylamine method. ResultsCompared with the SAP group, the levels of serum amylase, creatinine, blood urea nitrogen, IL-1β, IL-6, the cell apoptosis index, and the expression of iNOS mRNA were significantly decreased, the serum level of IL-10 and the activity of SOD were significantly increased (P < 0.05), the pancreas and the kidney damaged more slightly in the Curcumin group and Curcumin combined with Rhodiola group.Compared with the Curcumin group, the above situations were more better in the Curcumin combined with Rhodiola group. ConclusionsCurcumin combined with Rhodiola has a better protective effect on SAP associated renal injury.It might be through inhibiting the expressions of IL-1β, IL-6, stimulating the expression of IL-10, down-regulating the expression of iNOS mRNA, and improving the activity of SOD.It could reduce the cell apoptosis and necrosis of the kidney and improve the ability of the kidney to tolerate hypoxia.
Objective To investigate the effects and mechanism of curcumin on the retinal neovasularization in mice with oxygeninduced retinopathy (OIR). Methods A total of 72 C57BL/6J mice were divided into normal, OIR model, vehicle control ［dimethyl sulphoxide (DMSO)］, and curcumin group (100, 50, and 10 mg). The mice in normal group lived in normoxia condition; OIR model was set up according to standard methods in the literature. Five days after OIR establishment, the mice in curcumin group received an intraperitoneal (IP) injection of 0.1 ml curcumin (100, 50, and 10 mg), and the mice in DMSO group received an IP injection of 0.1 ml 1permil; DMSO. All of the mice were executed at the age of postnatal day 17 (P17) and the eyeballs were collected. Endothelial cell nuclei breaking through the internal limiting membrane were counted after stained with hematoxylin and eosin (HE). The expression of vascular endothelial growth factor-A (VEGF-A), vascular endothelial growth factor receptor-2 (VEGFR-2), endostatin (ES), and phosphorylated p38 mitogen-activated protein kinase (p-p38MAPK) in the retina in each group were measured by real-time polymerase chain reaction (RT-PCR) and Western blot methods.Results Compared with the normal group, retinal neovascularization was found in OIR model group (P＜0.05). The number of endothelial cell nuclei was 46.00plusmn;16.00 in OIR model group and 0.17plusmn;0.41 in normal group (P＜0.05). The expression of VEGF-A, ES, and p-p38MAPK in 100 mg curcumin group differed statistically from which in 50 and 10 mg curcumin group (P＜0.05). The expression of VEGFR-2 was same in the three curcumin groups (P＞0.05). Conclusion Curcumin can inhibit the formation of retinal neovascularization; the mechanism may be associated with inhibiting the expression of VEGFA and VEGFR-2, increasing the expression of ES, and inhibiting the p38MAPK signal transduction pathway.