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...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^(-1), SPD), alpha-difluoromethylornithine(2.5 mmol·L^(-1), DFMO), 4-Aminopyridine-(40 μ-mol·L^(-1), 4-AP), the polysaccharide extracts of RA or RAM(50, 100, or 200 mg·L^(-1)), DFMO plus SPD, or DFMO plus polysaccharide extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca^(2+)([Ca^(2+)]cyt) 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^(2+)]cyt 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^(2+)]cyt, 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 展开更多
[Objectives] To investigate the infrared identification of ZhuQin Formula and Atractylodes polysaccharides and its effect on immune function in immunocompromised mice. [Methods] In this experiment,the structure of pol...[Objectives] To investigate the infrared identification of ZhuQin Formula and Atractylodes polysaccharides and its effect on immune function in immunocompromised mice. [Methods] In this experiment,the structure of polysaccharides was analyzed by Fourier transform infrared spectroscopy. The experiment was divided into the polysaccharide groups of high,medium and low dose of ZhuQin,the extraction group of ZhuQin,the polysaccharide group of Atractylodes,the positive drug group,the blank group and the model group. The model of immunosuppressed mice was established by intraperitoneal injection of cyclophosphamide. The carbon clearance index,immune organ index,serum hemolysin,spleen T and B lymphocyte transformation rate were determined. [Results]The results show that the polysaccharides have obvious absorption peaks of polysaccharides; Formula extracts and polysaccharides can enhance the immune function of immunosuppressive mice at a certain dose level,and the effect of formula polysaccharide is better than that of extract and single traditional Chinese medicine polysaccharides. [Conclusions] The results suggest that the polysaccharides of ZhuQin Formula and Atractylodes may be acidic polysaccharides containing pyran ring and furan ring. ZhuQin Formula polysaccharide may play an immunomodulatory role mainly by improving the specific immunity of the organism.展开更多
基金supported by the National Natural Science Foundation of China(Nos.30772753,81173254,and 81673940)the Science and Technology Program of Guangzhou(No.20160701335)the First-class discipline construction major project of Guangzhou University of Chinese Medicine(Guangzhou University of Chinese Medicine Planning,2018-No.6)
文摘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^(-1), SPD), alpha-difluoromethylornithine(2.5 mmol·L^(-1), DFMO), 4-Aminopyridine-(40 μ-mol·L^(-1), 4-AP), the polysaccharide extracts of RA or RAM(50, 100, or 200 mg·L^(-1)), DFMO plus SPD, or DFMO plus polysaccharide extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca^(2+)([Ca^(2+)]cyt) 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^(2+)]cyt 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^(2+)]cyt, 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
基金Supported by Public Welfare and Industry Special Fund Project of the Ministry of Agriculture(201303040-05)Natural Science Foundation Project of CQCSTC(2013FYF110600)
文摘[Objectives] To investigate the infrared identification of ZhuQin Formula and Atractylodes polysaccharides and its effect on immune function in immunocompromised mice. [Methods] In this experiment,the structure of polysaccharides was analyzed by Fourier transform infrared spectroscopy. The experiment was divided into the polysaccharide groups of high,medium and low dose of ZhuQin,the extraction group of ZhuQin,the polysaccharide group of Atractylodes,the positive drug group,the blank group and the model group. The model of immunosuppressed mice was established by intraperitoneal injection of cyclophosphamide. The carbon clearance index,immune organ index,serum hemolysin,spleen T and B lymphocyte transformation rate were determined. [Results]The results show that the polysaccharides have obvious absorption peaks of polysaccharides; Formula extracts and polysaccharides can enhance the immune function of immunosuppressive mice at a certain dose level,and the effect of formula polysaccharide is better than that of extract and single traditional Chinese medicine polysaccharides. [Conclusions] The results suggest that the polysaccharides of ZhuQin Formula and Atractylodes may be acidic polysaccharides containing pyran ring and furan ring. ZhuQin Formula polysaccharide may play an immunomodulatory role mainly by improving the specific immunity of the organism.
