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1.
Article in Chinese | WPRIM | ID: wpr-1027493

ABSTRACT

Objective:To evaluate the effect of hyperthermia on radiation pneumonitis (RP) in elderly patients with esophageal cancer undergoing intensity-modulated radiotherapy (IMRT).Methods:Clinical data of 177 elderly esophageal cancer patients (aged ≥60 years) receiving IMRT in the First Affiliated Hospital of Soochow University and Yixing Cancer Hospital from August 1, 2017 to February 6, 2023 were retrospectively analyzed. Patients were divided into the hyperthermia and non-hyperthermia groups based on whether they received hyperthermia treatment. Patients in two groups received IMRT with 6 MV X-rays. Patients in the hyperthermia group underwent high-frequency hyperthermia within 1 h before radiation using the external thermotherapy device HG-2000Ⅲ (heating temperature: 41-43 ℃ for 40 min, twice a week). After adjusting for confounding factors between two groups using propensity score matching (PSM), the short-term effective rates between two groups were compared using Chi-square test. Univariate analysis and logistic multivariate analysis were employed to compare the incidence of RP between two groups. Results:After applying PSM, 42 pairs were successfully matched, and the baseline data and radiotherapy parameters showed no statistically significant differences between two groups (all P>0.05). The objective response rate (ORR) in the hyperthermia group was significantly higher than that in the non-hyperthermia group (83.3% vs. 64.3%, P=0.047). Univariate analysis revealed that the incidence of RP and symptomatic RP (≥ grade 2) in the hyperthermia group was significantly lower than that in the non-hyperthermia group (61.9% vs. 85.7%, P=0.013; 21.4% vs. 47.6%, P=0.012). Logistic multivariate analysis indicated that hyperthermia was an independent protective factor for symptomatic RP ( P=0.011). Conclusions:The incidence and severity of RP in elderly esophageal cancer patients receiving IMRT can be reduced by hyperthermia. Hyperthermia, as a clinically beneficial green treatment, improves efficacy and reduces toxicity for patients with esophageal cancer.

2.
China Pharmacy ; (12): 298-302, 2023.
Article in Chinese | WPRIM | ID: wpr-961662

ABSTRACT

OBJECTIVE To establish the fingerprint of Sophora flavescens, and to screen differential components and determine their contents. METHODS HPLC fingerprints of 12 batches of S. flavescens were established by using Similarity Evaluation System of Chromatographic Fingerprints of TCM (2012 edition); common peaks were identified and their similarities were evaluated. Chemical pattern recognition analysis [cluster analysis (CA),principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA)] were performed with SIMCA 14.1 and SPSS 23.0 software, and differential components which influenced the quality of S. flavescens were screen with variable importance in the projection(VIP)>1 as standard. Meanwhile, the contents of 4 kinds of differential components were determined by the same HPLC method. RESULTS There were 17 common peaks in the fingerprints of 12 batches of S. flavescens,and their similarities were all higher than 0.96. A total of 6 common peaks were identified, i.e. oxymatrine (peak 1), oxysophocarpine (peak 2), matrine (peak 10), trifolirhizin (peak 14), kurarinone (peak 16) and norkurarinone (peak 17). Results of CA, PCA and OPLS-DA showed that 12 batches of S. flavescens were divided into 3 categories according to producing area, i.e. S1-S7 (Shangzhou District of Shaanxi Province) were grouped into one category, S8-S10 (Yichuan County of Henan Province) into one category and S11-S12 (Chifeng City of Inner Mongolia) into one category. VIPs of matrine, norkurarinone, kurarinone and oxysophocarpine and the chemical components represented by peak 11 and 9 were all greater than 1. The contents of matrine, norkurarinone, kurarinone and oxysophocarpine in 12 batches of S. flavescens were 2.65-4.93, 1.54-3.44, 9.63-12.94 and 5.08-6.10 mg/g, respectively. CONCLUSIONS HPLC fingerprint of S. flavescens is established successfully in the study, and can be used to screen 6 differential components by combining with chemical pattern recognition analysis, which can provide reference for quality control of S. flavescens.

3.
Chinese Journal of Anesthesiology ; (12): 1380-1384, 2022.
Article in Chinese | WPRIM | ID: wpr-994122

ABSTRACT

Objective:To evaluate role of Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil protein kinase 2 (ROCK2) signaling pathway in trilobatin-induced reduction of cerebral ischemia-reperfusion (I/R) injury in rats.Methods:Eighty clean-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 230-280 g, were divided into 4 groups ( n=20 each) using a random number table method: sham operation group (group S), cerebral I/R group (group IR), cerebral I/R plus trilobatin group (group T) and cerebral I/R plus trilobatin plus RhoA/ROCK2 signaling pathway agonist AA group (group A). The model of focal cerebral I/R injury was developed by middle cerebral artery occlusion in anesthetized animals.Trilobatin 15 mg/kg was given by gavage twice a day for 3 consecutive days in T and A groups.RhoA/ROCK2 signaling pathway agonist AA 10 mg/kg was intraperitoneally injected before each administration by gavage in group A. Neurobehavioral score was assessed at 24 h of reperfusion, then the rats were sacrificed, and the hippocampal tissues were isolated for determination of the apoptosis rate of hippocampal neurons (by flow cytometry), cerebral infarction volume (by TTC staining), and expression of phosphorylated RhoA (p-RhoA), ROCK2 and cleaved caspase-3 (by Western blot) and for microscopic examination of ultrastructure of hippocampal neurons (with a transmission electron microscope). Results:Compared with group S, the neurobehavioral score, apoptosis rate of hippocampal neurons, and cerebral infarction volume were significantly increased, the expression of p-RhoA, ROCK2 and cleaved caspase-3 was up-regulated ( P<0.05), and the pathological damage to hippocampal neurons was aggravated in group IR.Compared with group IR, the neurobehavioral score, apoptosis rate of hippocampal neurons and cerebral infarction volume were significantly decreased, the expression of p-RhoA, ROCK2 and cleaved caspase-3 was down-regulated ( P<0.05), and the pathological damage to hippocampal neurons was alleviated in group T. Compared with group T, the neurobehavioral score, apoptosis rate of hippocampal neurons, and cerebral infarction volume were significantly increased, the expression of p-RhoA, ROCK2 and cleaved caspase-3 was up-regulated ( P<0.05), and the pathological damage to hippocampal neurons was aggravated in group A. Conclusions:RhoA/ROCK2 signaling pathway is involved in trilobatin-induced reduction of cerebral I/R injury, which may be related to inhibition of apoptosis in hippocampal neurons of rats.

4.
Article in Chinese | WPRIM | ID: wpr-957179

ABSTRACT

Objective:To construct and validate a nomogram model based on clinical factors and PET/CT metabolic parameters of 18F-FDG for predicting epidermal growth factor receptor (EGFR) mutations in lung adenocarcinoma. Methods:From January 2014 to January 2019, 114 patients (59 males, 55 females, age (60.0±10.8) years) with lung adenocarcinoma in the First Affiliated Hospital of Harbin Medical University were retrospectively enrolled. Clinical data (smoking status, tumor location, clinical stage and carcinoembryonic antigen (CEA) level), 18F-FDG PET/CT metabolic parameters (SUV max, metabolic tumor volume (MTV) and total lesion glycolysis (TLG)) and EGFR mutation status were analyzed. Patients were divided into training group (80 cases) and validation group (34 cases). In the training group, univariate analyses (independent-sample t test, Wilcoxon rank sum test, χ2 test or Fisher′s exact probability method) were used for categorical variables. Variables that showed significant differences between EGFR mutation group and wild type group were selected. Variance inflation factors (VIF) were calculated and the collinearity variables were deleted, and a nomogram model of optimal logistic model was constructed based on Akaike information criterion (AIC). The effect of the model was evaluated by the concordance index (C-index), sensitivity, specificity, accuracy, calibration and decision curve analysis (DCA) in the training group and the validation group. Results:Among 114 patients, 56 were with EGFR mutations and 58 were with EGFR wild type. In the training group, there were significant differences in gender (male/female: 14/26 vs 25/15; χ2=6.05, P=0.014), smoking status (with/without smoking history: 4/36 vs 22/18; χ2=18.46, P<0.001) and SUV max (5.72(3.90, 8.32) vs 8.09(4.56, 12.55); W=1 045.50, P=0.018) between EGFR mutation group and wild type group. However, there were no significant differences in other factors ( t=-0.54, χ2 values: 0.20 and 0.20, W values: 921.50 and 983.00, all P>0.05). The VIF of gender, smoking status and SUV max were all less than 10, and the nomogram model with three factors showed the minimum AIC (90.06). In the training group, C-index value of the model was 0.798 (95% CI: 0.699-0.897), with the sensitivity of 85.0%(34/40), the specificity of 70.0%(28/40) and the accuracy of 77.5%(62/80). In the validation group, C-index value was 0.854(95% CI: 0.725-0.984), with the sensitivity of 13/16, the specificity of 14/18, and the accuracy of 79.4%(27/34). The calibration curve and the goodness of fit test showed good calibration, and DCA showed that the model could benefit patients clinically within a large risk threshold range (training group: 0-0.59, validation group: 0-0.65). Conclusion:The nomogram model based on gender, smoking status and SUV max can be used to easily predict EGFR mutation status in lung adenocarcinoma.

5.
Article in Chinese | WPRIM | ID: wpr-1015078

ABSTRACT

AIM: To study the protective effect and mechanism of rosuvastatin on cerebral ischemia-reperfusion injury. METHODS: (1) Cerebral infarction and OGD/R cell models were established to detect the effects of different concentrations of rosuvastatin on cell proliferation and apoptosis; (2) Different concentrations of rosuvastatin were used to treat OGD/R cell models and to observe rosuvastatin effects on cell morphology and expression and localization of UCP2-SIRT3 in cells; (3) UCP2 silent cell line was constructed to observe cell mitochondrial morphology and expression and localization of TOMM20 and SIRT3 molecules in cells, and to study the channels and mechanisms that play a protective role of rosuvastatin in OGD/R cell model; (4) The mitochondrial membrane potential, mitochondrial gene PGC1, Drp1 and Opa1 expression were detected to study the protective effect of rosuvastatin on mitochondria. RESULTS: (1) Rosuvastatin of different concentrations could significantly reduce OGD/R cell apoptosis and increase cell survival rate; (2) Rosuvastatin exerted cell protection by affecting the expression of UCP2 and SIRT3 in cells, thereby protecting cells from OGD/R injury; (3) Rosuvastatin affected the expression of TOMM20 by regulating UCP2, increased mitochondrial transmembrane transport and energy metabolism, enhanced mitochondrial function, and improved cell state and reduced apoptosis. CONCLUSION: Rosuvastatin inhibits mitochondrial damage of OGD/R cells by regulating UCP2/SIRT pathway, thereby exerting neuron protection.

6.
Chinese Journal of School Health ; (12): 1022-1025, 2020.
Article in Chinese | WPRIM | ID: wpr-823169

ABSTRACT

Objective@#To explore the trajectory of mobile phone addiction score and to investigate the relationship between subgroups of trajectory and anxiety with depression in college students, and to provide evidence for risk factors of anxiety and depression and mobile phone addiction prevention college students.@*Methods@#A total of 1 562 college students were recruited from 2017 in Shandong University were followed longitudinally for five times by means of stratified cluster sampling. Mobile Phone Addiction Tendency Scale, Self-rating Anxiety Scale, and Self-rating Depression Scale were used, and latent class linear mixed models were used to identify the trajectory of mobile phone addiction score and Logistic regression models were used to explore the relationship between subgroups of trajectory with anxiety and depression.@*Results@#At the last survey, the mean mobile phone addiction score was (42.9±5.4) and the prevalence of anxiety and depression was 33.7% (n=526) and 40.2% (n=628), respectively. The trajectories of mobile phone addiction score were classified into five groups: stable, high level-decreasing group, low level-rapid increasing group, moderate level-increasing group, and high level-increasing group. The number and proportion of the five groups were 701(44.9%), 309(19.8%), 96(6.2%), 232(14.9%), 224(14.3%), respectively. Compared with students of stable group, students in the moderate level-increasing and high level-increasing groups had higher risk of anxiety (OR=3.19, 95%CI=2.32-4.40; OR=8.38, 95%CI=5.09-13.77) and depression (OR=3.29, 95%CI=2.40-4.52; OR=4.49, 95%CI=2.82-7.16).@*Conclusion@#Mental health education in universities should start with mobile phone intervention, especially those with severe increasing tendency of mobile phone addiction, which would subsequently decrease the prevalence of anxiety and depression in college students.

7.
Chinese Journal of Dermatology ; (12): 182-188, 2019.
Article in Chinese | WPRIM | ID: wpr-745761

ABSTRACT

Objective To assess the effect of fenretinide (4-HPR) on proliferation,apoptosis and migration of B16F10 and A375 melanoma cells,and to evaluate the effect of liposomes and RGD peptidemodified liposomes on its uptake and therapeutic effects.Methods A film-hydration method was used to prepare 4-HPR liposomes (4-HPRL),which were modified with RGD peptide to prepare RGD-4-HPRL,and the concentration,particle size,electric potential,drug loading capacity and encapsulation efficiency were measured for 4-HPRL and RGD-4-HPRL.In vitro cultured B16F10 and A375 cells were divided into several groups:4-HPR group,4-HPRL group and RGD-4-HPRL group treated with Dulbecco's minimum essential medium (DMEM) containing 4-HPR bulk drug,4-HPRL and RGD-4-HPRL respectively at the same concentration of 4-HPR,and control group treated with culture solution at the same volume.After different durations of treatment,cell counting kit-8 (CCK8) assay was performed to evaluate cellular proliferative activity,annexin V/propidium iodide staining to detect apoptosis,and scratch wound healing assay to evaluate the effect of drug treatment on cell migration ability.Then,4-HPR was replaced by coumarin 6 (C6) to prepare C6 liposomes (C6L) and RGD-C6L,and flow cytometry was conducted to evaluate C6 uptake by B16F10 cells.Statistical analysis was carried out with SPSS22.0 software by one-way analysis of variance (ANOVA) for the comparison among several groups and t test for the comparison between two groups.Results The concentration of 4-HPR in the prepared 4-HPRL solution was over 1 300 mg/L.The encapsulation efficiency and drug loading capacity of 4-HPRL were (95.51 ± 1.22)% and (7.27 ± 0.11)% respectively,and those of RGD-4-HPRL were (95.82 ± 0.81)% and (7.14 ± 0.13)% respectively.The particle size distribution of 4-HPRL and RGD-4-HPRL was uniform,and their average particle size was below 100 nm.CCK8 assay showed that 4-HPR could markedly inhibit the proliferative activities of B16F10 and A375 cells.The cell proliferation inhibition rate of 4-HPRL was higher than that of 4-HPR at the same concentration of 4-HPR (P < 0.01),and the inhibition rate of RGD-4-HPRL was higher than that of 4-HPRL (P < 0.01 or P < 0.05) and 4-HPR (P < 0.01).As annexin V/propidium iodide apoptosis assay showed,when the concentration of 4-HPR was 10 mg/L,the total apoptosis rates of B16F10 cells in the control group,4-HPR group,4-HPRL group and RGD-HPRL group were (4.44 ± 0.35)%,(28.33 ± 0.66)%,(46.43 ± 0.77)% and (51.33 ± 0.37)% respectively.When the concentration of 4-HPR was 20 mg/L,the total apoptosis rates of A375 cells in the above 4 groups were (4.97 ± 0.62)%,(16.68 ± 3.81)%,(32.62 ± 1.24)% and (44.85 ± 4.92)% respectively.The apoptosis rates of B16F10 and A375 cells were significantly higher in the 4-HPRL group than in the 4-HPR group (both P < 0.01),and higher in the RGD-4-HPRL group than in the 4-HPRL group (both P < 0.01) and 4-HPR group (both P <0.01).Scratch wound healing assay showed that 4-HPR could inhibit scratch healing and migration of B16F10 and A375 cells,and the inhibitory effects of 4-HPRL and RGD-4-HPRL were distinctly superior to those of 4-HPR bulk drug.C6 uptake assay revealed that the fluorescence intensity of C6 in B16F10 cells in the control group,C6 group,C6L group and RGD-C6L group were 2.15 ± 0.28,8.56 ± 0.36,20.48 ± 0.13 and 22.55 ± 0.07 respectively,and there were significant differences between the 4 groups (F =67 194.186,P < 0.01).Additionally,the fluorescence intensity of C6 was significantly higher in the C6L group and RGD-C6L group than in the C6 group (both P < 0.01),and higher in the RGD-C6L group than in the C6L Group (P < 0.01).Conclusions 4-HPR can inhibit the proliferation and migration of A375 and B16F10 cells,and induce their apoptosis.Liposomes and RGD-targeted liposomes can markedly enhance the effect of 4-HPR on melanoma cells.

8.
Chinese Journal of Burns ; (6): 683-689, 2018.
Article in Chinese | WPRIM | ID: wpr-807513

ABSTRACT

Objective@#To explore the effects of N-(4-hydroxyphenyl) retinamide (4HPR), 4HPR liposome (4HPR-L), and 4HPR lipid microbubble (4HPR-LM) combined with ultrasound on proliferation, apoptosis, and cell cycle of human keloid fibroblasts (Fbs).@*Methods@#(1) 4HPR-L and 4HPR-LM were prepared by hydration ultrasonic method. The appearance morphology, particle size distribution, Zeta potential, loading drug concentration, encapsulation efficiency, and drug loading rate of 4HPR-L were investigated by high performance liquid chromatography, dynamic light scattering, and transmission electron microscope. (2) Human keloid Fbs were cultured and divided into 13 groups by random number table (the same grouping method below), with 6 wells in each group. Cells in control group were given no treatment, while cells in 12 ultrasound groups including 0.5 W 30 s group, 0.5 W 60 s group, 0.5 W 120 s group, 0.7 W 30 s group, 0.7 W 60 s group, 0.7 W 120 s group, 1.0 W 30 s group, 1.0 W 60 s group, 1.0 W 120 s group, 1.5 W 30 s group, 1.5 W 60 s group, and 1.5 W 120 s group were treated by ultrasound with corresponding parameters. The cells viability was measured by a microplate reader after 24 hours of routine culture. Another batch of human keloid Fbs were divided into 5 groups, with 6 wells in each group. Cells in control group were given no treatment, while cells in 1, 10, 20, and 50 μg/mL blank lipid microbubble groups were treated with blank lipid microbubbles in corresponding mass concentration. The cells viability was measured as before after 24 hours of routine culture. Another batch of human keloid Fbs were divided into 6 groups, with 12 wells in each group. Cells in control group were given no treatment, while cells in 1, 10, 20, 50, and 100 μg/mL 4HPR-L groups were added with 4HPR-L carrying corresponding mass concentration of 4HPR. The cells viability in 6 wells of each group was detected after 24 and 48 hours of routine culture, respectively. Another batch of human keloid Fbs were divided into 4 groups, with 6 wells in each group. Cells in control group were given no treatment, while cells in 4HPR, 4HPR-L, and 4HPR-LM+ ultrasound groups were treated with 4HPR, 4HPR-L, and 4HPR-LM (all the mass concentration of 4HPR was 20 μg/mL), respectively, and cells in 4HPR-LM+ ultrasound group were given 0.5 W 60 s ultrasound treatment immediately after drug administration. The cells viability was measured as before after 24 hours of routine culture. (3) Another batch of human keloid Fbs were divided into control group, 4HPR group, 4HPR-L group and 4HPR-LM+ ultrasound group, with 3 wells in each group, and the cells in each group were treated as before. Apoptosis of the cells was detected by flow cytometer after 24 hours of routine culture. (4) Another batch of human keloid Fbs were grouped and treated as in (3), and then the cell cycle distribution was detected by flow cytometer after 24 hours of routine culture. Data were processed with one-way analysis of variance and t test.@*Results@#(1) 4HPR-L particles had a spherical or spheroidal structure and were uniform in size, with particle size of (100.1±1.3) nm and Zeta potential of (-34.3±2.3) mV. The mass concentration of 4HPR in 4HPR-L solution was about 1 400 μg/mL, with the encapsulation efficiency of (95.8±1.2)% and drug loading rate of (8.3±0.4)%. (2) The viability of cells in the 12 ultrasound groups was higher than 93.0%, and the viability of cells in 1, 10, 20, and 50 μg/mL blank lipid microbubble groups was higher than 95.0%. The viability of cells in 1 μg/mL 4HPR-L group at administration hour 24 was similar to that at 48 (t=0.393, P>0.05). The viability of cells in 10, 20, 50, and 100 μg/mL 4HPR-L groups at administration hour 24 was significantly higher than that at administration hour 48 (t=44.593, 22.961, 32.224, 35.337, P<0.01). The viability of cells in 4HPR group, 4HPR-L group, and 4HPR-LM+ ultrasound group was (47.3±0.7)%, (42.3±1.7)%, and (38.6±0.8)%, respectively. The viability of cells in 4HPR group was significantly higher than that in 4HPR-L group and 4HPR-LM+ ultrasound group (t=4.551, 15.895, P<0.05 or P<0.01). The viability of cells in 4HPR-L group was significantly higher than that in 4HPR-LM+ ultrasound group (t=-3.360, P<0.05). (3) The percentages of total apoptotic cells in 4HPR group, 4HPR-L group, and 4HPR-LM+ ultrasound group were (32.8±2.4)%, (42.5±2.4)%, and (58.5±6.3)%, respectively, which were significantly higher than the percentage of control group [(14.9±1.6)%, t=8.748, 13.637, 9.500, P<0.01]. The percentages of total apoptotic cells in 4HPR-L group and 4HPR-LM+ ultrasound group were significantly higher than the percentage in 4HPR group (t=4.049, 5.393, P<0.05 or P<0.01), and the percentage of total apoptotic cells in 4HPR-LM+ ultrasound group was significantly higher than that in 4HPR-L group (t=3.371, P<0.01). (4) The percentage of G2/M phase cells in 4HPR group was higher than that in control group, but there was no statistically significant difference (t=2.107, P>0.05). The percentage of G2/M phase cells in 4HPR-L group was significantly higher than that in 4HPR group or control group (t=18.169, 30.026, P<0.01). The percentage of G2/M phase cells in 4HPR-LM+ ultrasound group was significantly higher than that in 4HPR-L group, 4HPR group, and control group (t=4.932, 25.854, 66.231, P<0.01).@*Conclusions@#4HPR can inhibit proliferation, induce apoptosis, and arrest G2/M phase of human keloid Fbs, and the effects of 4HPR-LM combined with ultrasound are better than those of 4HPR-L and free 4HPR.

9.
Article in Chinese | WPRIM | ID: wpr-699685

ABSTRACT

Objective This study was to investigate the role of recombinant human platelet derived growth factor-BB(rhPDGF-BB) in the proliferation and migration of human retinal vascular endothelial cells (hRVECs).Methods hRVECs were cultured in DMEM with 10% fetal bovine serum.The rhPDGF-BB at the concentrations of 10,50 and 200 ng/ml were added into the medium of exponential phase-growth cells for 24 and 48 hours,respectively,and no rhPDGF-BB was added in the normal control group.The proliferation of the cells (absorbancy) was assayed by cell counting kit 8 (CCK8) method.Cell scratch test was employed to evaluate the relative migration area of cells (migrated acellular area/initial acellular area).The relative expression of rhPDGF-BB recepter (rhPDGF-BBR) mRNA in the cells was detected by reverse transcription PCR.The relative expression of VEGF mRNA and integrin mRNA in the cells was detected using real-time fluorescence quantitative PCR.Results hRVECs grew well and a expressing band according with rhPDGF-BBR prime was displayed.The absorbancy values of thecells were 1.01±0.05,1.09±0.04,1.10±0.02 and 1.13±0.05 in the normal control group and 10,50,200 ng/ml rhPDGF-BB groups at 24 hours after culture,and those in the 10,50 and 200 ng/ml rhPDGF-BB groups were significantly increased in comparison with the normal control group (t =2.504,3.430,3.483,all at P<0.05).The relative migrated areas of the cells in the normal control group and 10,50,200 ng/ml rhPDGF-BB groups were 0.42±0.10,0.38±0.09,0.55±0.06 and 0.61±0.05 at 24 hours after culture,and those at 48 hours were 0.75±0.06,0.81 ±0.02,0.87±0.02 and 0.98±0.02,showing significant differences among the groups (Fgroup =16.283,P =0.000;Ftime =209.129,P=0.000),and the relative migrated areas was depended upon the rhPDGF-BB dose and time.The relative expressions of integrin mRNA were 1.06 ± 0.02,1.30 ±0.10,1.20 ± 0.16 and 1.27 ± 0.08,and those of VEGF mRNA were 0.97±0.05,1.06±0.16,1.58 ±0.18 and 1.66 ±0.21 in the normal control group and 10,50 ng/ml,200 ng/ml rhPDGF-BB groups,respectively,and increased expressions of integrin mRNA and VEGF mRNA were found in the 50 and 200 ng/ml rhPDGF-BB groups compared with the normal control group (integrin mRNA:t =3.900,4.014,both at P < 0.05;VEGF mRNA:t =6.940,7.210,both at P < 0.05).Conclusions rhPDGF-BB/rhPDGF-BBR signal promotes the proliferation and migration of hRVECs probably by up-regulating the expressions of integrin and VEGF.

10.
Article in Chinese | WPRIM | ID: wpr-641116

ABSTRACT

Background The suppression of retinal angiogenesis is one of primary treatment targets for retinal vascular diseases,so seeking the intervention targets of retinal neovascularization is a hot research.Studies showed that insulin-like growth factor 1 (IGF-1) can promote the growth and restrain the apoptosis of vascular endothelial cells.However,whether IGF-1 is an intervention target for the treatment of retinal vascular diseases is unelucidated.Objective This study was to address the effects of IGF-1 on the migration,apoptosis and capillary tube formation of human retinal vascular endothelial cells (HRECs) and mechanism.Methods HRECs were cultured in vitro,and the cells in the exponential phase were prepared for subsequent experiments.The expression of IGF-1R mRNA in the cells was examined using reverse transcriptase PCR assay.Different concentrations of IGF-1 were added in the medium based on the difference of tests.The relative free-cell area difference (△S) after test was measured by Photoshop CS4 software and compared among 0,10 and 200 ng/ml IGF-1 groups 12 and 24 hours after cell scratching,respectively.The cell apoptotic rate was assayed by flow cytometry and compared between 0 ng/ml IGF-1 group and 1 000 ng/ml IGF-1 group,and the number of capillary tubes was examined by Matrigel test and assessed among 0,10,100 and 200 ng/ml IGF-1 groups 24 hours after addition of IGF-1.The expressions of platelet derived growth factor (PDGF)-BB mRNA and caspase-3 mRNA in the cells of the 0,500 and 1 000 ng/ml IGF-1 groups were detected by real-time fluorescence quantitative PCR after adding IGF-1 for 6 hours.Results Cultured cells grew well and attached 90% confluence 2-3 days after incubation,and IGF-1R mRNA was positively expressed in the cells.In 12 and 24 hours after scratching,the relative migrating area of the cells was gradually reduced with the increase of IGF-1 contents.The △S was (4.83 ± 0.61) × 105 μm2 in the 200 ng/ml IGF-1 group,which was significantly larger than (3.28±0.64) ×105 μm2 in the 0 ng/ml IGF-1 group 24 hours after stretching (t=-3.707,P=0.021).The apoptotic rate in the 0 ng/ml IGF-1 group and 1 000 ng/ml IGF-1 group was (18.77±2.37) % and (12.05 ±0.88) %,with a significant difference between them (t =2.869,P =0.046).The number of intact tubes was significantly increased in the 200 ng/ml IGF-1 group compared with the 0 ng/ml IGF-1 group ([20.33±2.83]/well vs.[17.94± 1.96]/well;t =-2.940,P =0.042).Compared with 0 ng/ml IGF-1 group,the relative expression level of PDGF-BB mRNA was elevated and that caspase-3 mRNA was evidently reduced in the 1 000 ng/ml IGF-1 group (t=-3.489,P =0.025;t =7.287,P =0.002).Conclusions IGF-1 can promote the migration and angiogenesis of HRECs and inhibit the apoptosis of HRECs.These effects of IGF-1 probably are associated with the up-regulation of PDGF-BB and down-regulation of caspase-3 in the cells.

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