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1.
Artigo em Chinês | WPRIM | ID: wpr-971523

RESUMO

OBJECTIVE@#To investigate the inhibitory effects of levofloxacin (LEV) combined with cellulase against bacille CalmetteGuerin (BCG) biofilms in vitro.@*METHODS@#The mature growth cycle of BCG biofilms was determined using the XTT method and crystal violet staining. BCG planktonic bacteria and BCG biofilms were treated with different concentrations of LEV and cellulose alone or jointly, and the changes in biofilm biomass were quantified with crystal violet staining. The mature BCG biofilm was then treated with cellulase alone for 24 h, and after staining with SYTO 9 and Calcofluor White Stain, the number of viable bacteria and the change in cellulose content in the biofilm were observed with confocal laser scanning microscopy. The structural changes of the treated biofilm were observed under scanning electron microscopy.@*RESULTS@#The MIC, MBC and MBEC values of LEV determined by broth microdilution method were 4 μg/mL, 8 μg/mL and 1024 μg/mL, respectively. The combined treatment with 1/4×MIC LEV and 2.56, 5.12 or 10.24 U/mL cellulase resulted in a significant reduction in biofilm biomass (P < 0.001). Cellulase treatments at the concentrations of 10.24, 5.12 and 2.56 U/mL all produced significant dispersion effects on mature BCG biofilms (P < 0.001).@*CONCLUSION@#LEV combined with cellulose can effectively eradicate BCG biofilm infections, suggesting the potential of glycoside hydrolase therapy for improving the efficacy of antibiotics against biofilmassociated infections caused by Mycobacterium tuberculosis.


Assuntos
Levofloxacino/farmacologia , Violeta Genciana/farmacologia , Vacina BCG/farmacologia , Antibacterianos/farmacologia , Biofilmes , Celulases/farmacologia , Testes de Sensibilidade Microbiana
2.
Artigo em Chinês | WPRIM | ID: wpr-1022407

RESUMO

Objective:To investigate the influencing factors for lymph node metastasis and prognosis in early gastric cancer.Methods:The retrospective cohort study was conducted. The clinicopathological data of 1 011 patients with early gastric cancer who were admitted to the Zhejiang Cancer Hospital from January 2010 to December 2019 were collected. There were 561 males and 450 females, aged (58±11)years. All patients underwent radical resection of gastric cancer and the lymph node metastasis of each group was identified according to the pathological examination on patients' surgical specimens. Observation indicators: (1) lymph node metastasis in early gastric cancer; (2) influencing factors for lymph node metastasis in early gastric cancer; (3) influencing factors for prognosis in early gastric cancer. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Comparison of ordinal data was analyzed using the non‐parameter rank sum test. Univariate analysis was conducted using the Log-Rank test and Logistic regression model, and multivariate analysis was conducted using the Logistic stepwise regression model and COX step-wise regression model. The Kaplan-Meier method was used to calculate survival rate and draw survival curve, and the Log-Rank test was used for survival analysis. Results:(1) Lymph node metastasis in early gastric cancer. The lymph node metastasis rate of 1 011 patients with early gastric cancer was 23.640%(239/1 011), in which the lymph node metastasis rate of patients with T1a stage gastric cancer was 11.883%(53/446), and the lymph node metastasis rate of patients with T1b stage gastric cancer was 32.920%(186/565). There were 239 patients with lymph node metastasis mainly concentrated in the first station, including 7 cases with No.1 lymph node metastasis, 11 cases with No.2 lymph node metastasis, 135 cases with No.3 lymph node metastasis, 59 cases with No.4 lymph node metastasis, 39 cases with No.5 lymph node metastasis, 91 cases with No.6 lymph node metastasis, 6 cases with No.7 lymph node metastasis, 8 cases with No.8 lymph node metastasis, 8 cases with No.9 lymph node metastasis and 6 cases with No.10 lymph node metastasis. Multiple lymph node metastases may exist in the same patient. For lymph node metastasis in different tumor sites, there were 4 cases, 2 cases and 1 case of lymph node metastasis in the No.2, 3 and 5 lymph node in patients with upper gastric cancer. There were 3 cases, 7 cases, 36 cases, 15 cases, 3 cases and 5 cases of lymph node metastasis in the No.1, 2, 3, 4, 5 and 6 lymph node in patients with middle gastric cancer. There were 4 cases, 97 cases, 44 cases, 35 cases and 86 cases of lymph node metastasis in the No.1, 3, 4, 5 and 6 lymph node in patients with lower gastric cancer. (2) Influencing factors for lymph node metastasis in early gastric cancer. Results of multivariate analysis showed that tumor diameter, tumor location, degree of tumor invasion, vascular thrombus, degree of tumor differentiation were independent factors influencing lymph node metastasis in early gastric cancer ( odds ratio=1.80, 1.49, 2.65, 5.76, 0.60, 95% confidence interval as 1.29-2.50, 1.11-2.00, 1.81-3.88, 3.87-8.59, 0.48-0.76, P<0.05). (3) Influencing factors for prognosis in early gastric cancer. All 1 011 patients were followed up for 43(range, 0-135)months, and the 3-year overall survival rate was 97.32%. Results of multivariate analysis showed that age >60 years and lymph node metastasis were independent risk factors influencing prognosis in early gastric cancer ( hazard ratio=9.50, 2.20, 95% confidence interval as 3.31-27.29, 1.00-4.87, P<0.05). Results of further analysis showed that the 3-year overall survival rate was 99.37% and 94.66% in patient with age >60 years and ≤60 years, respectively, showing a significant difference between them ( χ2=25.33, P<0.05). The 3-year overall survival rate was 95.42% and 97.92% in patients with and without lymph node metastasis, respectively, showing a significant difference between them ( χ2=5.69, P<0.05). Conclusions:The lymph node metastasis rate of early gastric cancer can reach 23.640%. Tumor diameter, tumor location, degree of tumor invasion, vascular thrombus, degree of tumor differentia-tion are independent factors influencing lymph node metastasis in early gastric cancer, age >60 years and lymph node metastasis are independent risk factors influencing prognosis.

3.
Chinese Journal of Urology ; (12): 181-186, 2022.
Artigo em Chinês | WPRIM | ID: wpr-933189

RESUMO

Objective:To explore independent risk factors and risk stratification for diagnosis of clinically significant prostate cancer (CsPCa) in biopsy-naive patients with nonsuspicious multiparametric magnetic resonance imaging (mpMRI).Methods:The data of 549 patients who underwent initial systematic biopsy (SB) in the First Affiliated Hospital of Soochow University, the Second Affiliated Hospital of Soochow University and Traditional Chinese Medicine Hospital of Kunshan between October 2015 and January 2021 were retrospectively reviewed. Nonsuspicious mpMRI was defined as Prostate Imaging-Reporting and Data System (PI-RADS)≤2. All patients received systematic 12 core prostate biopsy, 278 of them by transperineal and 271 by transrectal biopsies. The median age of the patients was 67 (62, 73) years, the median prostate specific antigen (PSA) was 9.01 (6.15, 13.64) ng/ml, the median prostate volume was 48.41 (35.85, 64.28) ml, and 54 patients were positive in digital rectal examination (DRE). Taking CsPCa as the outcome index, receiver operating characteristic (ROC) analysis was performed on age, tPSA, f/tPSA and PSA density (PSAD) to obtain the optimal cut-off value, and logistics regression was used to explore the independent risk factor of CsPCa in mpMRI negative patients. The optimal cut-off value when the negative predictive value (NPV) of mpMRI diagnosis of CsPCa was 100%, was taken as the protective factor, and the risk stratification model was finally proposed.Results:Of all 549 cases, 44 were CsPCa, 35 were clinically insignificant prostate cancer and 470 were non-prostate cancer. There were significant differences in age (71 vs. 67 years old), tPSA (11.95 vs. 8.75 ng/ml), PSAD [0.31 vs. 0.18 ng/(ml·cm 3)], f/tPSA (0.12 vs. 0.16) and DRE positive rate (38.6% vs. 7.3%) between CsPCa group and non-CsPCa group ( P<0.01). Cut-off values were taken in ROC analysis when the Youden index was at its maximum. The optimal cut-off values of each continuous variable were: age=65 years, tPSA=10ng/ml, f/tPSA=0.2 and PSAD=0.15 ng/(ml·cm 3). Multivariate analysis showed that ages over 65 years ( OR=3.43, 95% CI 1.55-7.58, P=0.002), f/t PSA ratio<0.2 ( OR=3.84, 95% CI 1.28-11.56, P=0.016), PSAD>0.15 ng/(ml·cm 3) ( OR=3.60, 95% CI 1.13-11.51, P=0.03) and positive DRE ( OR=5.20, 95% CI 2.39-11.32, P<0.001) were independent risk factors of CsPCa. When NPV was 100%, the cut-off values were taken as the protective factors: age≤55 years, f/tPSA≥0.3, PSAD≤0.1 ng/(ml·cm 3). Combined with independent risk factors, preliminary risk stratification was conducted: those with ≥2 high risk factors were considered as high risk group, those with ≥2 protective factors were considered as low risk group, and the middle region was considered as medium risk group. Conclusions:Patients with age>65 years, f/tPSA<0.2, PSAD > 0.15 ng/(ml·cm 3) and DRE positive are independent risk factors of CsPCa in mpMRI negative patients. Patients in the high-risk group were recommended to undergo prostate biopsy, while patients in the low-risk group could be considered to avoid biopsy.

4.
Artigo em Chinês | WPRIM | ID: wpr-936101

RESUMO

Objective: To investigate the functional outcomes and postoperative complications of Cheng's GIRAFFE reconstruction after proximal gastrectomy. Methods: A descriptive case series study was conducted. Clinical data of 100 patients with adenocarcinoma of the esophagogastric junction who underwent Cheng's GIRAFFE reconstruction after proximal gastrectomy in Cancer Hospital of University of Chinese Academy of Sciences (64 cases), Zhejiang Provincial Hospital of Chinese Medicine (24 cases), Lishui Central Hospital (10 cases), Huzhou Central Hospital (1 case) and Ningbo Lihuili Hospital (1 case) from September 2017 to June 2021 were retrospectively analyzed. Of 100 patients, 64 were males and 36 were females; the mean age was (61.3 ± 11.1) years and the BMI was (22.7±11.1) kg/m(2). For TNM stage, 68 patients were stage IA, 24 were stage IIA and 8 were stage IIB. Postoperative functional results and postoperative complications of radical gastrectomy with Giraffe reconstruction were analyzed and summarized. Gastroesophageal reflux disease questionnaire (RDQ) score and postoperative endoscopy were used to evaluate the occurrence of reflux esophagitis and its grade (grade N, grade A, grade B, grade C, and grade D from mild to severe reflux). The continuous data conforming to normal distribution were expressed as (mean ± standard deviation), and those with skewed distribution were presented as median (Q1, Q3). Results: All the 100 patients successfully completed R0 resection, including 77 patients undergoing laparoscopic surgery and 23 patients undergoing laparotomy. The Giraffe anastomosis time was (38.6±14.0) min; the blood loss was (73.0±18.4) ml; the postoperative hospital stay was 9.5 (8.2, 13.0) d; the hospitalization cost was (6.0±0.3) ten thousand yuan. Fourteen cases developed perioperative complications (14.0%), including 7 cases of pleural effusion or pneumonia, 3 cases of anastomotic leakage, 2 cases of gastric emptying disorder, 1 case of gastrointestinal hemorrhage and 1 case of anastomotic stenosis, who were all improved and discharged after symptomatic management. Patients were followed up for (33.3±1.6) months. Eight patients were found to have reflux symptoms by RDQ scale six months after surgery, and 11 patients (11/100,11.0%) were found to have reflux esophagitis by gastroscopy, including 6 in grade A, 3 in grade B, and 2 in grade C. All the patients could control their reflux symptoms with behavioral guidance or oral PPIs. Conclusion: Cheng's GIRAFFE reconstruction has good anti-reflux efficacy and gastric emptying function; it can be one of the choices of reconstruction methods after proximal gastrectomy.


Assuntos
Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adenocarcinoma/cirurgia , Neoplasias Esofágicas/cirurgia , Esofagite Péptica/etiologia , Junção Esofagogástrica/cirurgia , Gastrectomia/métodos , Refluxo Gastroesofágico/etiologia , Laparoscopia , Procedimentos de Cirurgia Plástica/métodos , Recuperação de Função Fisiológica , Estudos Retrospectivos , Neoplasias Gástricas/cirurgia
5.
Artigo em Chinês | WPRIM | ID: wpr-799568

RESUMO

Objective@#To investigate the safety and feasibility of proximal partial gastrectomy with Cheng's Giraffe esophagogastric reconstruction for the treatment of early Siewert II adenocarcinoma of esophagogastric junction (AEG).@*Methods@#Indication of Cheng's Giraffe esophagogastric reconstruction: (1) Siewert II AEG or Siewert III AEG with diameter < 4 cm; (2) preoperative staging as cT1-2N0M0. A descriptive case series study was carried out. Clinical data of 34 patients with Siewert II AEG undergoing proximal partial gastrectomy and Cheng's Giraffe esophagogastric reconstruction at Department of Abdominal Surgery of Zhejiang Cancer Hospital and Department of Gastrointestinal Surgery, The First Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine from February to July 2018 were retrospectively collected and analyzed, including 14 cases in IA stage, 11 cases in IIA stage and 8 cases in IIB stage. Brief procedure of Cheng's Giraffe esophagogastric reconstruction was as follows: Firstly, 12 cm long tubular stomach was formed by longitudinal incision 4 cm away from the great curvature of the stomach. Secondly, the gastric fundus and His angle were formed. Finally, the distance from His angle to esophagal-tubular gastric anastomosis should be more than 5 cm. The reflux disease questionare (RDQ) scores, radionuclide gastric emptying scintigraphy, and 24-hour multichannel intraluminal (MII)-pH monitoring technology were used to evaluate postoperative gastric emptying and gastroesophageal reflux.@*Result@#All 34 patients successfully completed proximal partial gastrectomy with Cheng's Giraffe esophagogastric reconstruction, including 13 cases by open surgery and 21 cases by laparoscopic surgery. The operation time was (144.6±39.8) minutes, the blood loss during operation was (35.4±17.2) ml. No laparoscopic case was converted to open surgery and no postoperative complication was observed. The postoperative hospital stay was (8.4±2.5) days. The postoperative RDQ score was 4.4±3.1 one month after operation, and 3.3±2.5 six months after operation. Gastric-half emptying time was (67.0±21.5) minutes, and the residual ratio was (52.2±7.7)% in 1 hour, (36.4±3.1)% in 2 hours and (28.8±3.6)% in 3 hours at postoperative 1-month. The 24-hour MII-pH monitoring at postoperative 2-month revealed the frequency of acid reflux was (12.6±7.9) times, frequency of non-acid reflux was (19.6±9.7) times, DeMeester score was 5.8±2.9.@*Conclusion@#Cheng's Giraffe esophagogastric reconstruction is safe and feasible in the treatment of Siewert type II AEG, and has good dynamic and anti-reflux effects.

6.
Artigo em Inglês | WPRIM | ID: wpr-880585

RESUMO

OBJECTIVES@#Contrast-induced acute kidney injury (CI-AKI) is the third cause of hospital-acquired AKI, and existing clinical prevention and treatment measures such as hydration therapy and/or administration of antioxidants N-acetylcysteine treatment and other treatments still show little effect on the prevention and treatment of CI-AKI. This study aims to explore the effect of Danhong injection on prevention of CI-AKI.@*METHODS@#A total of 12 867 patients, who received coronary angiography, percutaneous coronary intervention, enhanced CT or vascular intervention in a tertiary hospital, were enrolled for this study. Among them, 423 in the treatment group received intravenous drip of Danhong injection, and 12 444 in the control group received routine medicine. Propensity score matching was conducted to balance confounding factors between the 2 groups and then the prevention effect of Danhong injection on CI-AKI was compared between them.@*RESULTS@#A total of 423 pairs of patients were matched successfully. The incidence of CI-AKI in the non-Danhong control group was higher than that in the Danhong treatment group (5.7% vs 2.4%). The difference between the 2 groups was statistically significant (@*CONCLUSIONS@#The results of this study support the use of Danhong injection in the prevention of the Stage 1 of CI-AKI.


Assuntos
Humanos , Injúria Renal Aguda/prevenção & controle , Meios de Contraste/efeitos adversos , Medicamentos de Ervas Chinesas , Injeções , Intervenção Coronária Percutânea , Pontuação de Propensão , Fatores de Risco , Resultado do Tratamento
7.
Acta Pharmaceutica Sinica B ; (6): 690-701, 2019.
Artigo em Inglês | WPRIM | ID: wpr-774951

RESUMO

Oxidative stress and cardiomyocyte apoptosis are involved in the pathogenesis of doxorubicin (DOX)-induced cardiotoxicity. Matrine is well-known for its powerful anti-oxidant and anti-apoptotic capacities. Our present study aimed to investigate the effect of matrine on DOX-induced cardiotoxicity and try to unearth the underlying mechanisms. Mice were exposed with DOX to generate DOX-induced cardiotoxicity or normal saline as control. H9C2 cells were used to verify the effect of matrine . DOX injection triggered increased generation of reactive oxygen species (ROS) and excessive cardiomyocyte apoptosis, which were significantly mitigated by matrine. Mechanistically, we found that matrine ameliorated DOX-induced uncoupling protein 2 (UCP2) downregulation, and UCP2 inhibition by genipin could blunt the protective effect of matrine on DOX-induced oxidative stress and cardiomyocyte apoptosis. Besides, 5'-AMP-activated protein kinase 2 () deficiency inhibited matrine-mediated UCP2 preservation and abolished the beneficial effect of matrine in mice. Besides, we observed that matrine incubation alleviated DOX-induced H9C2 cells apoptosis and oxidative stress level activating AMPK/UCP2, which were blunted by either AMPK or UCP2 inhibition with genetic or pharmacological methods. Matrine attenuated oxidative stress and cardiomyocyte apoptosis in DOX-induced cardiotoxicity maintaining AMPK/UCP2 pathway, and it might be a promising therapeutic agent for the treatment of DOX-induced cardiotoxicity.

8.
Artigo em Chinês | WPRIM | ID: wpr-775058

RESUMO

OBJECTIVE@#To study the changes and significance of apoptosis signal-regulating kinase 1 (ASK1) in left ventricular remodeling in FVB/N mice.@*METHODS@#A total of 54 FVB/N mice were randomly divided into 4 groups: 0 d group with 8 mice, 7 d group with 10 mice, 14 d group with 16 mice, and 21 d group with 20 mice. A model of cardiac remodeling was established by intraperitoneal injection of isoproterenol (ISO) at a daily dose of 30 mg/kg, and the 7 d, 14 d, and 21 d groups were injected for 7, 14, and 21 consecutive days respectively. The 0 d group was given intraperitoneal injection of an equal volume of normal saline. Echocardiography was used to measure left ventricular posterior wall thickness at end diastole (dLVPW) and the ratio of heart weight to tibia length (HW/TL) was measured. Hematoxylin-eosin staining was used to measure left ventricular myocardial fiber diameter. Picric-Sirius red staining was used to measure myocardial collagen deposition area in the left ventricle. Quantitative real-time PCR was used to measure the mRNA expression of ASK1, type I collagen (collagen I), and B-type natriuretic peptide (BNP). The mortality rate was observed for each group.@*RESULTS@#There were gradual increases in HW/TL, myocardial fiber diameter, and dLVPW after 0, 7, and 14 days of ISO injection (P0.05), while there was a significant reduction in myocardial fiber diameter (P0.05). There were significant increases in myocardial collagen deposition area and the mRNA expression of collagen I, ASK1, and BNP after 0, 7, 14, and 21 days of ISO injection, which reached the peaks on day 21 (P<0.01). The mRNA expression of ASK1 was positively correlated with myocardial collagen deposition area and the mRNA expression of collagen I and BNP and had a weak correlation with HW/TL, myocardial fiber diameter, and dLVPW. There was a significant increase in the mortality rate of the mice over the time of ISO injection.@*CONCLUSIONS@#The expression of ASK1 in the myocardium is closely associated with left ventricular remodeling. The increase of ASK1 expression may lead to the aggravation of left ventricular remodeling, and the mechanism of which needs further study.


Assuntos
Animais , Camundongos , Isoproterenol , MAP Quinase Quinase Quinase 5 , Miocárdio , Miócitos Cardíacos , Remodelação Ventricular
9.
Chinese Pharmacological Bulletin ; (12): 844-849, 2019.
Artigo em Chinês | WPRIM | ID: wpr-857237

RESUMO

Aim: To explore the mechanism of the effects of sanguinarine on the proliferation, invasion and cell cycle of gastric cancer cells. Methods: MTT assay was used to detect the inhibitory effect of sangui-narine(3, 6, 9 μmol · L-1) on the cell viability of gastric cancer SGC-7901 cells. Wound healing and flow cytometry were applied to detect cell migration and cell cycle respectively. Western blot was employed to determine the protein expression levels of migration and cell cycle related proteins MMP-2, MMP-9, cyclin Dl, CDK4, and pi 6. Results: Compared with control group, the cell viability decreased significantly with the increase of drug concentration after 24 h treatment (P < 0. 05). Flow cytometry results showed that the proportion of cells in Gl phase after treatment with 3, 6, 9 μmol · L-1 of sanguinarine significantly increased (P < 0. 05). Wound healing results indicated that the migration ability of gastric cancer cells was significantly inhibited after sanguinarine treatment. After sanguinarine treatment, the protein expression levels of MMP-2, MMP-9, cyclin Dl and CDK4 were significantly down-regulated (P < 0. 05), whereas the protein expression level of pi 6 was significandy up-regulated (P < 0. 05). Conclusions: Sanguinarine significantly inhibits the proliferation, reduces the migration ability of cells, induces cell arrest in Gl phase, and its anticancer effect is related to its regulation of MMP-2, MMP-9, cyclin Dl, CDK4 and p16.

10.
Artigo em Inglês | WPRIM | ID: wpr-773572

RESUMO

Astragalus membranaceus (Radix Astragali, RA) and Atractylodes macrocephala (Rhizoma Atractylodis Macrocephalae, RAM) are often used to treat gastrointestinal diseases. In the present study, we determined the effects of polysaccharides extracts from these two herbs on IEC-6 cell migration and explored the potential underlying mechanisms. A migration model with IEC-6 cells was induced using a single-edged razor blade along the diameter of cell layers in six-well polystyrene plates. The cells were grown in control media or media containing spermidine (5 μmol·L, SPD), alpha-difluoromethylornithine (2.5 mmol·L, DFMO), 4-Aminopyridine (40 μmol·L, 4-AP), the polysaccharide extracts of RA or RAM (50, 100, or 200 mg·L), DFMO plus SPD, or DFMO plus polysaccharide extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca ([Ca]) was measured using laser confocal microscopy, and cellular polyamine content was quantified with HPLC. Kv1.1 mRNA expression was assessed using RT-qPCR and Kv1.1 and RhoA protein expressions were measured with Western blotting analysis. A cell migration assay was carried out using Image-Pro Plus software. In addition, GC-MS was introduced to analyze the monosaccharide composition of both polysaccharide extracts. The resutls showed that treatment with polysaccharide extracts of RA or RAM significantly increased cellular polyamine content, elevated [Ca] and accelerated migration of IEC-6 cells, compared with the controls (P < 0.01). Polysaccharide extracts not only reversed the inhibitory effects of DFMO on cellular polyamine content and [Ca], but also restored IEC-6 cell migration to control level (P < 0.01 or < 0.05). Kv1.1 mRNA and protein expressions were increased (P < 0.05) after polysaccharide extract treatment in polyamine-deficient IEC-6 cells and RhoA protein expression was increased. Molar ratios of D-ribose, D-arabinose, L-rhamnose, D-mannose, D-glucose, and D-galactose was 1.0 : 14.1 : 0.3 : 19.9 : 181.3 : 6.3 in RA and 1.0 : 4.3 : 0.1 : 5.7 : 2.8 : 2.2 in RAM. In conclusion, treatment with RA and RAM polysaccharide extracts stimulated migration of intestinal epithelial cells via a polyamine-Kv1.1 channel activated signaling pathway, which facilitated intestinal injury healing.


Assuntos
Animais , Ratos , Astragalus propinquus , Química , Atractylodes , Química , Linhagem Celular , Movimento Celular , Medicamentos de Ervas Chinesas , Química , Farmacologia , Células Epiteliais , Biologia Celular , Metabolismo , Intestinos , Biologia Celular , Genética , Metabolismo , Poliaminas , Metabolismo , Polissacarídeos , Química , Farmacologia , Rizoma , Química , Transdução de Sinais , Proteína rhoA de Ligação ao GTP , Metabolismo
11.
Artigo em Inglês | WPRIM | ID: wpr-812361

RESUMO

Astragalus membranaceus (Radix Astragali, RA) and Atractylodes macrocephala (Rhizoma Atractylodis Macrocephalae, RAM) are often used to treat gastrointestinal diseases. In the present study, we determined the effects of polysaccharides extracts from these two herbs on IEC-6 cell migration and explored the potential underlying mechanisms. A migration model with IEC-6 cells was induced using a single-edged razor blade along the diameter of cell layers in six-well polystyrene plates. The cells were grown in control media or media containing spermidine (5 μmol·L, SPD), alpha-difluoromethylornithine (2.5 mmol·L, DFMO), 4-Aminopyridine (40 μmol·L, 4-AP), the polysaccharide extracts of RA or RAM (50, 100, or 200 mg·L), DFMO plus SPD, or DFMO plus polysaccharide extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca ([Ca]) was measured using laser confocal microscopy, and cellular polyamine content was quantified with HPLC. Kv1.1 mRNA expression was assessed using RT-qPCR and Kv1.1 and RhoA protein expressions were measured with Western blotting analysis. A cell migration assay was carried out using Image-Pro Plus software. In addition, GC-MS was introduced to analyze the monosaccharide composition of both polysaccharide extracts. The resutls showed that treatment with polysaccharide extracts of RA or RAM significantly increased cellular polyamine content, elevated [Ca] and accelerated migration of IEC-6 cells, compared with the controls (P < 0.01). Polysaccharide extracts not only reversed the inhibitory effects of DFMO on cellular polyamine content and [Ca], but also restored IEC-6 cell migration to control level (P < 0.01 or < 0.05). Kv1.1 mRNA and protein expressions were increased (P < 0.05) after polysaccharide extract treatment in polyamine-deficient IEC-6 cells and RhoA protein expression was increased. Molar ratios of D-ribose, D-arabinose, L-rhamnose, D-mannose, D-glucose, and D-galactose was 1.0 : 14.1 : 0.3 : 19.9 : 181.3 : 6.3 in RA and 1.0 : 4.3 : 0.1 : 5.7 : 2.8 : 2.2 in RAM. In conclusion, treatment with RA and RAM polysaccharide extracts stimulated migration of intestinal epithelial cells via a polyamine-Kv1.1 channel activated signaling pathway, which facilitated intestinal injury healing.


Assuntos
Animais , Ratos , Astragalus propinquus , Química , Atractylodes , Química , Linhagem Celular , Movimento Celular , Medicamentos de Ervas Chinesas , Química , Farmacologia , Células Epiteliais , Biologia Celular , Metabolismo , Intestinos , Biologia Celular , Genética , Metabolismo , Poliaminas , Metabolismo , Polissacarídeos , Química , Farmacologia , Rizoma , Química , Transdução de Sinais , Proteína rhoA de Ligação ao GTP , Metabolismo
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