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OBJECTIVE To investigate the inhibitory effects of Ginkgo biloba extract (GBE) on renal inflammation in diabetic nephropathy (DN) model mice, and its potential mechanism. METHODS KK/Ay mice were fed with high fat and high sugar to induce DN model. They were divided into model group, positive control group [metformin 200 mg/(kg·d)], GBE low-dose and high-dose groups [100, 200 mg/(kg·d)], with 6 mice in each group. Six C57BL/6J mice were fed with a regular diet as the control group. Administration groups were given relevant liquid intragastrically, control group and model group were given constant volume of normal saline intragastrically, once a day, for 8 consecutive weeks. The body weight, fasting blood glucose, 24-hour food intake, 24-hour urine output, monocyte chemoattractant protein-1 (MCP-1), interleukin-12 (IL-12), IL-10, advanced glycation end products (AGEs), blood urea nitrogen (BUN) and serum creatinine (Scr) of mice were measured, and the ratio of bilateral kidneys to body weight was also calculated. The pathological injury and fibrotic changes of the renal cortex were observed, and the expressions of macrophage polarization marker proteins [type M1: inducible nitric oxide synthase (iNOS); type M2: arginase-1 (Arg-1)] and AGEs-the receptor of advanced glycation end products (RAGE)/Ras homolog gene pharm_chenjing@163.com family member A (RhoA)/Rho-associated coiled-coil forming protein kinase (ROCK) signaling pathway-related proteins were determined in renal cortex. RESULTS Compared with the model group, the symptoms such as renal cortical hyperplasia, vacuoles, infiltration of inflammatory cells, and renal cortical fibrosis had been improved in GBE low-dose and high-dose groups; body weight, serum level of IL-10, the expression of Arg-1 in the renal cortex were significantly higher than model group (P< 0.01); fasting blood glucose, 24-hour food intake, 24-hour urine output, serum levels of MCP-1, IL-12, BUN, Scr and AGEs, the ratio of bilateral kidneys to body weight, renal injury score, the proportion of renal interstitial fibrosis, the protein expressions of iNOS, RAGE, RhoA and ROCK1 (except for GBE low-dose group) in renal cortex were significantly lower than model group (P<0.01). CONCLUSIONS GBE could improve kidney damage and alleviate inflammatory response in DN model mice, the mechanism of which may be related to inhibiting the AGEs-RAGE/RhoA/ROCK signaling pathway and regulating macrophage polarization.
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OBJECTIVE To investigate the protective effect and potential mechanism of cornuside on diabetic nephropathy (DN) model mice. METHODS Male KK-Ay mice were fed with high-fat and high-sugar diet for two weeks to reproduce the DN model. The successfully modeled mice were randomly grouped into model group, aminoguanidine group (positive control,100 mg/kg) and cornuside group (100 mg/kg), and male C57BL/6J mice were included as normal group, with 6 mice in each group. Administration groups were given relevant medicine intragastrically, and normal group and model group were given a constant volume of normal saline intragastrically, once a day, for 8 consecutive weeks. The levels of fasting blood glucose (FBG), 24 h urinary protein, serum interleukin-12 (IL-12), IL-10, blood urea nitrogen (BUN) and serum creatinine (Scr) were detected; the pathological injury, fibrotic change and glomerular microstructure of renal tissue were observed; the expressions of the receptor of advanced glycation end products (RAGE), collagen type Ⅳ (COL-Ⅳ) and inducible nitric oxide synthase (iNOS) in renal cortex were detected in each group. RESULTS Compared with normal group, the renal cortex of mice in model group showed obvious inflammatory cell infiltration and fibrotic changes; the mesangial hyperplasia of glomerulus was serious and the basement membrane had a large number of irregular dark dense deposits; the levels of FBG and 24 h urinary protein, the serum levels of IL- 12, BUN and Scr, and the expression levels of RAGE, COL-Ⅳ and iNOS in the renal cortex were significantly increased, while the serum level of IL-10 was significantly decreased (P<0.01). Compared with the model group, the renal pathological injuries, fibrotic changes and glomerular microstructure of mice in administration groups were improved significantly, and the above quantitative indexes were generally improved (P<0.05 or P<0.01). CONCLUSIONS Cornuside has a certain protective effect on DN model mice. It can inhibit the inflammatory response, reduce urinary protein excretion, and alleviate renal fibrosis, which may be related to the inhibition of the advanced glycation end products/RAGE signaling pathway.
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Fibroblast growth factors (FGF) are a group of structurally related polypeptides which constitute an elaborate signaling system with their receptors. Evidence accumulated in the years suggests that the FGF family plays a key role in the repair of central nervous system injury. The main protective mechanisms include activating the expression of PI3K-Akt, peroxisome proliferator-activated receptor (PPARγ) and other signals; inhibiting NF-κB-mediated inflammatory response, oxidative stress and apoptosis; regulating neuronal differentiation and neuronal excitability as well as participating in protection of neurovascular units and nerve function repair. This paper comprehensively summarizes the latest research progress in FGF signaling related to diseases of the central nervous system such as cerebral infarction, cerebral hemorrhage, traumatic brain injury, Alzheimer's disease, Parkinson's disease, epilepsy and depression, aiming to provide scientific basis and reference for the development of innovative FGF drugs for the prevention and treatment of neurological diseases.
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Humans , Fibroblast Growth Factors , Phosphatidylinositol 3-Kinases/metabolism , Central Nervous System/metabolism , Signal Transduction/physiology , Alzheimer DiseaseABSTRACT
Objective:To investigate the effect of using two different input functions to reconstruct 18F-FDG PET/CT Patlak multi-parameter images on the quantitative parameters of lung cancer lesions. Methods:The original whole-body dynamic 18F-FDG PET/CT scan data of lung cancer patients in the Department of Nuclear Medicine, First Affiliated Hospital of Anhui Medical University were retrospectively analyzed. The total scan time was 75 min. Two input functions were used for Patlak multi-parameter reconstruction: ① Image-derived input function(IDIF)using the Time-activity curve(TAC)of descending aorta from 0 min to 75 min. ② Population-based input function (PBIF) developed by Yale University. Metabolic rate of FDG (MR FDG) and Distribution volume (DV) images were obtained by Patlak multi-parameter analysis software using the above input functions. The region of interest (ROI) method was used to delineate the lesions to obtain multi-parameter quantitative information, including the max, peak and mean value of MR FDG and DV. Paired t-test was used for statistical analysis. Results:The original data of 27 lung cancer patients who received whole-body dynamic 18F-FDG PET/CT imaging were reconstructed by Patlak with two different input functions. The max, peak and mean values of MR FDG-IDIF and MR FDG-PBIF in lung cancer lesions were as follows: (0.26 ± 0.15), (0.19 ± 0.12), (0.14 ± 0.08)μmol·min -1·ml -1 and (0.26 ± 0.15), ( 0.20 ± 0.13), (0.15 ± 0.09)μmol·min -1·ml -1, with no statistically significant difference between two functions( P > 0.05). The max, peak and mean values of DV IDIF and DV PBIF were (165.56 ± 99.89)%, (117.66 ± 72.24)%, (62.16 ± 33.65)% and(170.04 ± 103.93)%, (121.91 ± 73.71)%, (65.05 ± 37.17)%, with no statistically significant difference between two functions ( P > 0.05). Conclusions:The population-based input function may be an alternative for patients who could not lie supine for long time during whole-body dynamic 18F-FDG PET/CT Patlak multi-parameter imaging.
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Objective:To investigate the radiosensitizing effect of silencing breast cancer susceptibility gene 1 (shBRCA1) expression on MDA-MB-231 breast cancer bearing nude mice by 3′-deoxy-3′- 18F-fluorothymidine ( 18F-FLT) microPET/CT imaging. Methods:Twenty-four BALB/c nude mice were divided into 4 groups ( n=6 in each group) according to the random number table method, namely negative control (NC) group, NC+ radiotherapy group, shBRCA1 group and shBRCA1+ radiotherapy group. 18F-FLT microPET/CT imaging was performed before and 24 h after radiotherapy. The changes of SUV max before and after radiotherapy were compared among 4 groups, and the total proliferative volume (TPV) of tumors in each group after treatment was also analyzed. The expression of Ki-67 in tumor tissues was analyzed by immunohistochemistry. Data were analyzed by paired t test, one-way analysis of variance, least significant difference t test and Pearson correlation analysis. Results:Breast cancer cells targeting the BRCA1 were constructed. Before radiotherapy, the differences of SUV max among the NC group, NC+ radiotherapy group, shBRCA1 group and shBRCA1+ radiotherapy group were not statistically significant (1.034±0.137, 1.031±0.152, 1.028±0.169 and 1.026±0.156; F=0.00, P=0.999). Twenty-four hours after the end of the four times of radiotherapy, the differences of SUV max among the 4 groups were statistically significant (1.367±0.100, 0.781±0.079, 1.306±0.213 and 0.597±0.129; F=44.77, P<0.001), with lower SUV max in the shBRCA1+ radiotherapy group compared with the NC+ radiotherapy group ( t=2.98, P=0.014). The SUV max of the NC+ radiotherapy group and shBRCA1+ radiotherapy group were reduced compared with those before radiotherapy ( t values: 5.82, 5.44, P values: 0.002, 0.003), while SUV max of the NC group and shBRCA1 group increased compared with those before radiotherapy ( t values: -4.47, -3.98, P values: 0.007, 0.011). TPV was smaller in the shBRCA1+ radiotherapy group compared with that in the NC+ radiotherapy group (0.48±0.03 vs 0.61±0.07; F=25.36, t=3.82, P=0.003). Immunohistochemical assays showed that Ki-67 was less expressed in the shBRCA1+ radiotherapy group than that in the NC+ radiotherapy group (0.286±0.072 vs 0.476±0.093; F=15.73, t=3.61, P=0.007). Correlation analysis showed a positive correlation between Ki-67 expression and SUV max ( r=0.83, P<0.001). Conclusion:18F-FLT microPET/CT imaging can evaluate the radiosensitizing effect of shBRCA1 expression on MDA-MB-231 breast cancer bearing nude mice.
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Objective To investigate the effect and mechanism of Liuling Jiedu Pills on acute pharyngitis caused by Staphylococcus aureus in rats.Methods The rat model of acute pharyngitis was replicated using the method of injecting 1×109 CFU·mL-1 of Staphylococcus aureus solution into the pharynx of rats.SD rats were randomly divided into a blank group,a model group,a Lanqin Oral Solution group(5 mL·kg-1),and a low-,medium-,and high-dose group of Liuling Jiedu Pills(4.375,8.750,and 17.500 mg·kg-1),with 10 rats in each group.Rats in each group were administered the drug by gavage once a day for 7 days.The general conditions of the rats were observed and recorded every day during the modeling and drug administration periods,and the local inflammation in the pharynx was scored;histopathological changes in the pharynx of the rats were observed by hematoxylin-eosin(HE)staining;serum interleukin 1β(IL-1β),interleukin 6(IL-6),tumor necrosis factor α(TNF-α),and tumor necrosis factor-α(TNF-α)were detected by ELISA.Immunohistochemistry and Western Blot were used to detect the protein expression levels of IL-1β,IL-6 and TNF-α in rat pharyngeal tissue.Results Compared with the blank group,rats in the model group had significantly increased pharyngeal erythema,significantly higher inflammation scores(P<0.01),significantly lower body mass on days 5-7 after modeling(P<0.05,P<0.01),significantly higher pathological scores(P<0.01),significantly higher levels of the serum inflammatory factors IL-1β,IL-6,and TNF-α(P<0.01),and significantly higher pharyngeal tissues showed significantly higher levels of IL-1β,IL-6,and TNF-α proteins(P<0.01).Compared with the model group,the pharyngeal erythema was significantly reduced in the Lanqin Oral Solution group and the low-,medium-and high-dose groups of Liuling Jiedu Pills,and the inflammation scores were significantly reduced(P<0.01),and the serum levels of IL-1β,IL-6,and TNF-α were significantly reduced(P<0.01);the body mass of the rats in the Lanqin Oral Solution group,and in the medium-and high-dose groups of Liuling Jiedu Pills,were significantly increased on the seventh day of the modeling(P<0.01);the histopathological scores and the levels of IL-1β,IL-6 and TNF-α proteins in pharyngeal tissue were significantly decreased(P<0.05,P<0.01).Conclusion Liuling Jiedu Pills can significantly improve the symptoms and inflammatory pathological changes of pharyngeal tissues in rats with acute pharyngitis,and its mechanism may be related to the down-regulation of the expression levels of inflammatory factors such as IL-1β,IL-6,and TNF-α.
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OBJECTIVE To study the improvement effects and its mechanism of catalpol on testicular lesions in KK-Ay spontaneous diabetic mice on the basis of glycolysis process mediated by advanced glycation end products (AGEs) and their receptors (RAGE). METHODS KK-Ay spontaneous diabetic mice fed with high-fat diet were used as diabetic model, and then randomly divided into model group, catalpol group (100 mg/kg), aminoguanidine group (AGEs inhibitor, 100 mg/kg) and FPS- ZM1 group (RAGE inhibitor, 1 mg/kg), and C57BL/6J mice fed in the same period were set as normal group, with 6 mice in each group. The catalpol group and aminoguanidine group mice were given relevant medicine intragastrically, normal group and model group mice were given constant volume of normal saline intragastrically, and FPS-ZM1 group mice were given relevant medicine 1 mL/g intraperitoneally, for consecutive 8 weeks. After the last administration, the body mass, fasting blood glucose, 24-hour food intake, water consumption, urine volume, testicular organ coefficient, and sperm motility of the mice were measured; pathological morphology and ultrastructural structure of testicular tissue were observed; the levels of reduced glutathione (GSH), superoxide dismutase (SOD), lactate dehydrogenase (LDH) and sugar metabolites in testicular tissue of mice were detected; pathway enrichment analysis was performed; the level of AGEs in serum and testicular tissue, protein expressions of RAGE, B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax), and mRNA expressions of key rate-limiting enzymes [hexokinase (HK), phosphofructose kinase (PFK), pyruvate kinase (PK), LDH] in testicular tissue were alldetected. RESULT S Catalpol could significantly improve the general symptoms, testicular organ coefficients and motility ofsperm in KK-Ay spontaneous diabetic mice (P<0.05 or P<0.01). The morphology and ultrastructure of spermatogenic cells in each layer of the seminiferous tubules were all improved. The levels of GSH, SOD and LDH in testicular tissue,the levels of the metabolic product glucose fructose-1,6-diphosphate, 3-phosphate glycerate, 3-phosphate glyceraldehyde, lactic acid and pyruvate, the expressions of HK, PFK, PK and LDH mRNA were all significantly increased(P<0.05 or P<0.01); the levels of AGEs in serum and testicular tissue, the expression of RAGE protein and the ratio of Bax to Bcl-2 in testicular tissue were significantly decreased(P<0.05 or P<0.01). Aminoguanidine and FPS-ZM1 could significantly improve the levels of most of above indicators in mice(P<0.05 or P<0.01). CONCLUSIONS Catalpol shows significant improvement effects on testicular lesions of KK-Ay spontaneous diabetic mice, and its mechanism of action was associated with upregulation of AGEs/RAGE signaling pathway- mediated glycolysis.
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Objective:To investigate the value of 18F-FDG total-body PET/CT dynamic imaging in evaluating the efficacy of early radiotherapy for MDA-MB-231 breast cancer bearing nude mice. Methods:MDA-MB-231 breast cancer bearing nude mice were established and divided into control group and radiotherapy group based on the random number table method ( n=10 for each group). 18F-FDG total-body PET/CT dynamic imaging was performed before and after the radiotherapy. The SUV max and the maximum tracer uptake net influx constant ( Kimax) of tumors, and the target-to-background ratio (TBR) were calculated. The change of tumor volume was recorded. The value of 18F-FDG total-body PET/CT dynamic imaging in evaluating the efficacy of radiotherapy was accessed using pathological findings as the reference. Paired t test, independent-sample t test and Pearson correlation analysis were used for data analysis. Results:After radiotherapy, SUV max and Kimax(4.66±0.46 and 0.14±0.03) were reduced in the radiotherapy group compared with those before radiotherapy (5.30±0.52 and 0.19±0.03; t values: 4.61, 8.31, P values: 0.001, <0.001), while the SUV max (5.94±0.74 vs 5.24±0.50) and Kimax (0.23±0.03 vs 0.19±0.02) were increased compared with baseline in the control group ( t values: 4.77, 6.87, P values: 0.001, <0.001). TBR Ki was significantly higher than TBR suv based on all images of the 2 groups (14.11±5.58 vs 5.91±1.60; t=8.92, P<0.001). The tumor volume in the radiotherapy group decreased compared with that before radiotherapy, but the difference was not statistically significant ((0.74±0.12) vs (0.81±0.08) cm 3; t=2.24, P=0.052). The results of immunohistochemistry showed that glucose transport protein (Glut)1 was highly expressed in tumors, and the Glut1 positive cell percentage of the radiotherapy group was significantly lower than that of the control group ((38.30±6.18)% vs (69.78±5.37)%; t=12.17, P<0.001). The expression of Glut1 was significantly positively correlated with SUV max and Kimax(the control group: rsuv=0.75, P=0.012; rKi=0.77, P=0.010; the radiotherapy group: rsuv=0.67, P=0.035; rKi=0.77, P=0.010). The apoptosis index (AI) of tumor cells in the radiotherapy group was higher than that in the control group ((24.15±4.00)% vs (10.15±3.05)%; t=8.85, P<0.001). Conclusion:18F-FDG total-body PET/CT dynamic imaging can sensitively monitor the effect of early radiotherapy in MDA-MB-231 breast cancer bearing nude mice.
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Objective To investigate the feasibility of sodium glycididazole(CMNa)on the radio-sensitization of esophageal cancer(EC)by 18F-fluoromisonidazole(FMISO)PET/CT.Methods A total of 60 patients with EC from December 2016 to December 2017 were enrolled prospectively and were divided into control group(n = 30;21 males,9 females;age:(56.6±10.0)years)and experimental group(n=30;20 males,10 females;age:(59.3±9.0)years)using completely randomized grouping design.Patients in the control group received radiotherapy alone,and those in the experimental group were treated with conventional radiotherapy and CMNa.All patients underwent 18F-FMISO PET/CT imaging 1 week before and after radiotherapy.The imaging results were visually and semi-quantitatively analyzed.The maximum standardized uptake value(SUVmax),tumor/muscle ratio(T/M),and hypoxia volume(HV)were calculated.Paired t test,two-sample t test and/x2 test were used to analyze the data.Results T/M and HV in the control group before and after radiotherapy were 3.92±0.57 vs 1.66±0.35,(1.84±0.31)vs(1.04±0.15)mm3,respectively;T/M and HV in the experimental group before and after radiotherapy were 4.01±0.68 vs 1.27±0.11,(2.01±0.22)vs(0.90±0.09)mm3,respectively.The primary tumor T/M,HV after radiotherapy in 2 groups were significantly lower than those before radiotherapy(t values:12.15-24.43,all P<0.05)and the amplitudes of T/M and HV in the experimental group were significantly higher than those in the control group(?T/M:2.77±0.60vs 2.25±0.49,?HV:(1.12±0.18)vs(0.81±0.26)mm3;t values:3.00 and 1.80,both P<0.05).Meanwhile,the local control rate of EC after 3 months in the experimental group was higher than that in the control group(80.0%(24/30)vs 53.3%(16/30);x2=4.80,P<0.05).Conclusion CMNa has the radiosensitizing effect on EC and the 18F-FMISO PET/CT can evaluate the radiosensitization effect.
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Objective To study the radiosensitization of irisquinone on the Warburg effect of MDA-MB231 cells.Methods MDA-MB231 cells in the exponential growth phase were divided into 6 groups:control group,irisquinone group,radiation group,irisquinone plus radiation group(Irisquinone + RA),negative control group,and experimental group (siRNA).Colony formation assay was used to measure cell survival fraction of MDA-MB231 cells.Single-hit multi-target model was used to fit the survival curve and calculate the sensitive enhancement ratio (SER).Flow cytometry was used to measure cell apoptosis.The relative expression levels of HK Ⅱ mRNA and protein in each group were detected by qRT-PCR and Western blot respectively.Results The values of D0,Dq and SF2 in the irisquinone plus radiation group were obviously lower than those in the radiation group.The SER of irisquinone was 1.52.Compared with the other groups,the cell apoptosis rate was increased (t =13.29,12.09,5.90,3.83,P < 0.05),while the relative expression levels of HK Ⅱ mRNA (t =9.14,10.48,3.40,P<0.05) and protein (t=13.39,16.08,5.81,P < 0.05) were decreased in the irisquinone plus radiation group significantly.Conclusions Radiosensitization function of irisquinone inhibits the Warburg effect of MDA-MB231 cells by down-regulating the expression of HK Ⅱ.
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Objective To investigate the effect of irisquinone (IR) on the radiosensitization of MDA-MB-231 breast cancer in nude mice using 18F-fluoroerythronitroimidazole (18F-FETNIM) microPET/ CT.Methods Thirty-two nude mice bearing MDA-MB-231 breast cancer were divided into 4 groups by random number table method (n =8 for each group):group A was treated with radiotherapy alone,group B was treated with radiotherapy and IR,group C was treated with IR alone,and group D was fed with distilled water.18F-FETNIM microPET/CT images were obtained to monitor the maximum standardized uptake value (SUVmax) of tumor before radiotherapy and 24 h after radiotherapy.The mice were sacrificed and tumor tissues were removed for HE staining.The expression of hypoxia inducible factor-1α (HIF-1α) was detected by immunohistochemical (IHC) method.Data were analyzed by paired t test,one-way analysis of variance,the least significant different t test and Pearson correlation analysis.Results There was no significant difference in SUVmax among the 4 groups before radiotherapy (1.429±0.090,1.430±0.076,1.445±0.071,1.432± 0.074;F=0.072,P>0.05).At 24 h after radiotherapy,the SUVmax of group A and B decreased significantly (1.075±0.098,0.890±0.076;t values:12.888,33.217,both P<0.05),and there was significant difference between group B and group A (t =4.197,P<0.05).However,the SUVmax of group C and D increased significantly (1.617±0.090,1.644±0.063;t values:-11.009,-16.061,both P<0.05).Pathological results showed that tumor cells in group A and B were reduced.IHC results showed that the expression of HIF-1α was lower in group B than others ((26.75±7.19)%;F=46.745,t values:2.898-9.743,all P< 0.05).The expression of HIF-1α was positively correlated with SUVmax(r =0.89,P<0.05).Conclusion 18F-FETNIM microPET/CT could evaluate the radiosensitization effect of IR on MDA-MB-231 breast cancer in nude mice.
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Aim To investigate the effect that catalpol intervenes macrophage polarization mediated by mouse mesangial cells(MMCs) stimulated by advanced glycation end products(AGEs).Methods RAW264.7 macrophages and MMCs were co-cultured in vitro and divided into model group(100 mg·L-1 AGEs), control group(100 mg·L-1 BSA), catalpol(0.1, 1.0, 10.0 μmol·L-1) group, and aminoguanidine(1.0 μmol·L-1) group which was set as positive control.After being incubated with catalpol for 1 h, MMCs were stimulated by AGEs for 23 h.The proliferation-inhibition rate of MMCs was measured by MTT assay.MCP-1 in supernatant liquid of MMCs was detected by ELISA method.The expression of iNOS, CD16/32, TNF-α, COX-2, CD206 and Arg-1 was detected by Western blot.Simultaneously, the percentage of iNOS and CD206 was also measured by flow cytometry.Results AGEs could increase the level of MCP-1 secreted by MMCs.The expression of iNOS, TNF-α, CD16/32 and COX-2 protein of macrophage was up-regulated after MMCs stimulated by AGEs, while the expression of CD206 and Arg-1 was down-regulated.After being intervened by catalpol, these effects could be reversed.All the changes were concentration-related.Conclusions Catalpol can inhibit macrophages M1-type polarization process and promote M2-type polarization, which may be mediated through MCP-1 secreted by MMCs after AGEs stimulation.Catalpol can ameliorate inflammation and relieve diabetic kidney injury.
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Aim To explore the protective effect of lo-ganin ( an active component in Cornus officinalis ) on podocyte injury induced by advanced glycation end products ( AGEs) and its possible mechanism. Meth-ods Mouse podocytes were cultured in vitro and di-vided into Normal group, model group ( AGEs group) , loganin group and aminoguanidine group ( set as posi-tive control) . After being incubated with loganin( final concentrations are 0. 1, 1, 10 μmol · L-1 ) for 1 h, podocytes were stimulated by AGEs of 100 mg · L-1 for 24 h. Then, the cell viability was measured by u-sing MTT method. Podocytes apoptosis was evaluated by Hoechst33342/PI staining and flow cytometry. Re-ceptors of advanced glycation end products ( RAGE ) ,desmin and apoptosis-related protein like Bax, Bcl-2, cleaved caspase-3 in podocytes were detected by Western blot. Results Loganin ameliorated podocyte injury induced by AGEs, down-regulated the expression of desmin and RAGE. Loganin also reduced the apoptotic rate of podocytes and decreased the ratio of Bax/ Bcl-2 and the expression of pro-apoptotic protein cleaved caspase-3 in podocytes. Conclusion Loganin could ameliorate podocyte injury, and its mechanism may be related to the decrease of the expression of RAGE and inhibition of the apoptotic pathway.
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Aim To observe the protective mechanism of loganinand morroniside ( active components in Cornus officinalis) on HUVEC injury induced by advanced glycation end products ( AGEs ) .Methods HUVECs were cultured in vitro and divided into control group , model group ( AGEs group ) , loganin group , morroni-side group and aminoguanidine group ( set as positive control).After being incubated with loganin and mor-roniside( final concentrations were 100,10,1 μmol?L-1 ) for 1 h, HUVECs were stimulated by AGEs of 200 mg? L-1 for 24 h.Then, the cell viability was measured by using MTT method .The supernatant was extracted and the levels of NO ,ET-1,MCP-1,VCAM-1 were measured by the corresponding kits .Receptors of advanced glycation end products ( RAGE ) and NF-κB in HUVEC were detected by Western blot .Results Loganin and morroniside could inhibit HUVEC injury induced by AGEs .In model group ,the contents of ET-1,MCP-1,VCAM-1 increased(P<0.01),the content of NO decreased ( P <0.01 ) and the expression of RAGE and NF-κB increased(P<0.01); however,lo-ganin and morronside could reduce the ET-1,MCP-1, VCAM-1contents,increase the NO content and down-regulate the expression of RAGE and NF-κB to differ-ent extents .Conclusion Loganin and morroniside could ameliorate HUVEC injury , and its mechanism may be related to inhibit inflammation , the improve-ment of endothelial cell function , and the decrease of the expression of RAGE .
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Objective To evaluate the feasibility of 18F-FDG PET/CT in assessing the radiosensitizing effect of oleanolic acid (OA) in C6 rat glioma model,and to explore the possible mechanism of radiosensitizing effect of OA.Methods Thirty-two C6 glioma-bearing SD rats were divided into 4 groups by random number table:group A without OA and radiotherapy,group B with OA alone,group C with radiotherapy alone,group D with OA and radiotherapy.18F-FDG PET/CT was performed before radiotherapy,at 24 h and 7 d after radiotherapy.The tumor SUVmax was measured.All rats were sacrificed and tumor tissues were excised for HE and immunohistochemistry staining.Paired t test,one-way analysis of variance,LSD-t test and Spearman correlation analysis were performed to analyze the data using SPSS 13.0.Results There was no statistically significant difference in SUVmax among the 4 groups (SUVmax of groups A,B,C,D:5.252± 0.536,5.261±0.544,5.273±0.520,5.232±0.507) before treatment (F=0.008,P>0.05).At 24 h postradiotherapy,SUVmax of groups C and D (4.766±0.511 and 4.403 ±0.486) decreased significantly (t =14..788,13.366,both P<0.05);while groups A and B (5.680±0.635 and 5.763±0.689) increased significantly (t =-11.578,-8.651,both P<0.05;F=10.550,P<0.05).However,there was no statistically significant difference between groups C and D (t =1.453,P>0.05).At 7 d post-radiotherapy,SUVmax of groups C and D decreased significantly (t =9.750,10.530,both P<0.05);while SUVmax of groups A and B (t=-35.353,-6.884,both P<0.05) increased significantly (F=97.691,P>0.05).SUVmax of Group D decreased significantly compared with that of group C (t =5.329,P>0.05).Less tumor cells and more areas of necrosis were observed in group D compared with those in other groups by HE staining.The expression of HIF-1α was lower in group D than that in other groups by immunohistochemistry staining.HIF-1α expression was positively correlated with SUVmax(r=0.853,P<0.05).Conclusion 18F-FDG PET/CT has potential to evaluate the radiosensitizing effect of OA in vivo,and the radiosensitization of OA might be related to the down regulation of HIF-1α.
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Aim To evaluate the reversal effect of 2-deoxy-D-glucose ( 2-DG ) on multidrug resistance ( MDR) by observing the uptake change of 99m Tc-MIBI in HNE-1/DDP cells, and to explore its mechanism. Methods The uptake of 99m Tc-MIBI in HNE-1/DDP cells under different concentrations of 2-DG was detec-ted by γ-counter, and the clearance rates of 99m Tc-MI-BI in HNE-1 cells and HNE-1/DDP cells after treated with 2-DG (10 mmol·L-1 ) were compared. The con-tent of ATP in HNE-1/DDP cells was detected after treated with 2-DG. P-glycoprotein ( P-gp ) and multi-drug resistance-associated proteins ( MRP ) expression were measured by Western blot. Apoptotic HNE-1/DDP cells treated with DDP alone or combined with 2-DG (10 mmol·L-1 ) were detected by propidium io-dide ( PI ) staining. Results The clearance rate of 99m Tc-MIBI in HNE-1/DDP cells was 54. 8%, which was significantly higher than that ( - 41. 3%) in HNE-1 cells (P<0. 01). The clearance rate of 99mTc-MIBI in HNE-1/DDP cells was -203. 7% after treat-ment with 2-DG ( 10 mmol · L-1 ) , which could be significantly reduced compared with the control group ( P<0. 01 ) . The level of ATP was 55 . 69% compared with the negative control group and the expression of P-gp and MRP protein decreased dramatically in HNE-1/DDP. With the combination of 2-DG and DDP, the ap-optotic rate of HNE-1/DDP cells reached 49 . 4%which was significantly higher than DDP treated group (22. 5%) . Conclusion Multidrug resistance and the reversal effect of 2-DG on multidrug resistance could be evaluated effectively by detecting the uptake change of 99m Tc-MIBI in HNE-1/DDP cells. The mechanism may be related with the inhibition of ATP level and the re-duced expression of P-gp and MRP protein in cancer cells.
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[ ABSTRACT] AIM:To study the effect of idazoxan ( IDA) on the permeability of blood-brain barrier ( BBB) and the expression of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) in mouse ex-perimental autoimmune encephalomyelitis (EAE).METHODS: Female C57BL/6 mice (n=36) were randomly divided into control group, EAE group and IDA group, with 12 mice in each group.EAE was induced by myelin oligodendrocyte glycoprotein 35-55 ( MOG35-55 ) .IDA (2 mg/kg, ip, bid) was administered for 15 d after immunization.The neurological defects of the mice were observed daily and scored.The pathological changes were observed under microscope with HE stai-ning and LFB myelin staining.The BBB permeability was detected by Evans blue extravasation.The expression of MMP-9 and TIMP-1 in the brain of EAE mice was determined by Western blotting.RESULTS: Compared with EAE group, the score of neurological defects in IDA group was decreased, the inflammation was relieved, the BBB permeability was re-duced, and the expression MMP-9 and the ratio of MMP-9/TIMP-1 were decreased ( P<0.05 ) .CONCLUSION: The neuroprotective effect of IDA on mouse EAE might be related to the down-regulation of MMP-9 and the ratio of MMP-9/TIMP-1, thus reducing the degradation of BBB and the permeability of BBB, and ameliorating the pathologic process of EAE.
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The protective effects of Da Chai Hu Granules (DCHKL) on islet cells which were incubated with 4 mmol x L(-1) alloxan (AXN) were studied. The viability of islet cells were measured with MTT. Insulin released into medium and in islets was detected by radioimmunoassay. Cell apoptosis rate was determined by flow cytometry. The expression of anti-apoptotic gene Bcl-2 and pro-apoptotic gene Bax in islet cells were measured with RT-PCR (reverse transcription polymerase chain reaction). Serum containing DCHKL can promote the activity of islet cells significantly (P < 0.01). Basal insulin secretion and high glucose-stimulated insulin secretion increased significantly (P < 0.01). Serum containing DCHKL can inhibit apoptosis of islet cells, the ratio of apoptosis was decreased. Serum containing DCHKL increased expression of Bcl-2 mRNA and decreased expression of Bax mRNA. DCHKL can significantly promote proliferation of islet cells and increase the amount of basal secretion of pancreatic islet cells and high glucose-stimulated insulin secretion. The expression of Bcl-2 increased significantly. The expression of Bax decreased significantly. DCHKL have a protective effect on the islet cells.
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Objective To evaluate the value of 18F-FDG PET/CT in assessing radiosensitivity enhancement by irisquinone (IR) on rabbit xenografted VX2 lung tumor models.Methods Twenty-four tumor-beating rabbits were randomly divided into 3 groups (8 rabbits/group):group A with radiotherapy alone,group B with combined radiotherapy and IR,and group C without radiotherapy (the control group).18F-FDG PET/CT imaging was performed before radiotherapy and 24 h and one week after radiotherapy.The tumor SUVmax on delayed imaging was calculated in all rabbits.Two rabbits in each group were sacrificed after PET/CT imaging.HE staining was used to assess the differences in cancer cells among groups.Paired t test,one-way analysis of variance and Kaplan-Meier analysis were performed to analyze the data using SPSS 13.0.Results Before radiotherapy,the tumor SUVmax of all the 24 rabbits on standard and delayed imaging were 2.200 ± 0.761 and 3.162 ± 0.833 (t =-5.582,P < 0.01).At 24 h post-radiotherapy,the delayed SUVmax of groups A,B and C were 2.614 ± 0.654,2.349 ± 0.869 and 5.663 ± 1.144,respectively.The differences between pre-radiotherapy and 24 h post-radiotherapy were statistically significant in all three groups (t =2.527,3.620,11.011,all P <0.05).One week after radiotherapy,the delayed SUVmax of groups A,B and C were 3.625 ± 1.064,3.058 ±0.850 and 7.424 ± 1.751,respectively.The differences among groups A,B and C were statistically significant (tA∶ B =2.652,tA∶C =3.799,tB∶C =4.366,all P <0.05).The cancer cells of group B were fewer than those of groups A and C by pathological findings,which was consistent with 18F-FDG PET/CT results.The survival times of groups A,B and C were (62.375 ±4.534),(69.000 ±4.660) and (54.125 ±5.276) d,respectively.Kaplan-Meier survival curves revealed better survival of group B as compared to groups A and C,respectively (Log-rank,x2 =7.355,16.943,both P < 0.01).Conclusion 18 F-FDG PET/CT is able to evaluate the effect of irisquinone on tumor radiosensitivity enhancement.
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Objective To investigate the event-based prospective memory (EBPM) and time-based prospective memory (TBPM) in temporal lobe epilepsy patients,and test the hypothcsis that temporal lobe is involved in the prospective memory network. Methods Sixty-two patients with temporal lobe epilepsy (TLE) and 30 age- and education level-matched healthy adults were examed with the neuropsychological battery of tests including EBPM and TRPM tasks. Results In comparison with the controls,the TLE patients were impaired on MMSE,DS and VFT.Compared with the healthy adults ( EBPM:6.83± 1.34,TBPM:5.00 ± 1.70),the patients with temporal lobe epilepsy had a significant loss in EBPM (3.95 ±2.77,t =6.72,P <0.01 ) and TBPM (3.08 ±2.42,t =4.39,P <0.01).There was no difference in the performance of EBPM (3.82 ± 2.70,4.10 ± 2.90 ; t =- 0.40,P > 0.05 ) and TBPM (2.55 ± 2.20,3.69± 2.55; t =-1.90,P >0.05 ) between the patients with taking antiepileptic drugs and those without antiepileptic drugs. Conclusions These results suggest that patients with temporal lobe epilepsy may experience general difficulties with prospective memory.It is possible that temporal lobe is involved in the prospective memory network.The passages of TBPM need more self-initiated processes.