1.Effect of chondroitin sulfate on the proliferation and differentiation of HL60 cells
Wenhua XU ; Hao YU ; Yinting ZHANG ; Yinlin GE ; Fangsong GENG
Chinese Journal of Tissue Engineering Research 2006;10(37):166-168,封三
BACKGROUND: Chondroitin sulfate is the important component of cell matrix, it can accelerate the proliferation of tumor cells and restrain its ransfer.OBJECTIVE: To observe the effects of chondroitin sulfate on the proliferation and differentiation of HL60 cells under the action of adriamycin.DESIGN: An open experiment.SETTING: Department of Biochemistry and Molecular Biology, Medical College of Qingdao University.MATERIALS: The experiments were carried out in the Research Room of Biochemistry and Molecular Biology, Medical College of Qingdao University of September 2003 to December 2004. Experimental materials and reagents: HL60 cell strains, which were the cells from promylocytic leukemia, were purchased from Shanghai Cell Bank, Chinese Academy of Medical Sciences; Bovine cartilage chondroitin sulfate (Sigma) was also used.METHODS: ① After the passage and culture, the cells at the logarithmic proliferative phase were dispensed into cell suspension of 1×108 L-1 with RPMI1640 culture medium containing inactivated fetal bovine serum of 0.1in volume fraction, and then filled into the culture bottles with 4 mL in each bottle for a total of 45 bottles. ② Chondroitin sulfate was added to 15 bottles filled with cell suspension according to the concentrations of 0, 5,25, 50 and 75 mg/L respectively, and 3 bottles for each concentration, and 0.01 mol/L phosphate buffer (pH7.2) was added in the blank control group. Then the density of HL60 cells was determined by cell counting after treatment of chondroitin sulfate. ③ Thirty bottles filled with cell suspension were divided into chondroitin sulfate+adriamycin group and chondroitin sulfate group, 15 bottles in each group. Chondroitin sulfate of 0, 5,25, 50 and 75 mg/L was added to the two groups, and 3 bottles for each concentration, and 0.01 mol/L phosphate buffer (pH7.2) wass adokd in the blank control group. Then the survival rate of chondroitin sulfate treated HL60 was detected after adding adriamycin. ④ Chondroitin sulfate was added to 15 bottles filled with cell suspension according to the concentrations of 0, 5, 25, 50 and 75 mg/L respectively, and 3 bottles for each concentration, 0.01 mol/L phosphate buffer (pH7.2) was added in the blank control group. The activity of acid phosphatase was detected with enzymelinked immunosorbant assay (ELISA) in each group, and the effect of the cell differentiation was observed.MAIN OUTCOME MEASURES: ① Effects of chondroitin sulfate on the proliferation of HL60 cells; ② Effects of chondroitin sulfate plus adriamycin on the survival rate of HL60 cells; ③ Effect of chondroitin sulfate on the activity of acid phosphatase of HL60 cells.RESULTS: Totally 45 bottles of cell suspension were prepared, and all were involved in the analysis of results. ① As compared with the blank control group, the densities of HL60 cells at 24 hours after treated with chondroitin sulfate of different concentrations were all significantly increased (P < 0.01), which were increased more obviously in the 50 and 75 mg/L chondroitin sulfate treated groups, and there was no significant difference between the two groups (P > 0.05), which indicated that chondroitin sulfate within the range of concentration did not accelerate the growth of cells greatly. ② As compared with the blank control group, the survival rates of HL60 cells in the chondroitin sulfate+adriamycin groups were decreased to different extents after chondroitin sulfate of different concentrations were added, and it decreased obviously when the concentration of chondroitin sulfate was higher than 25 mg/L (P < 0.01). ③ As compared with the blank control group, the A values of acid phosphatase of the HHL60 cells were all obviously increased in the 5, 25 and 50 mL chondroitin sulfate treated groups (1.268±0.038, 1.305±0.101, 1.321±0.021,1.354±0.013, P < 0.01 or 0.05), especially that it reached 1.406±0.113 in the 75 mL chondroitin sulfate treated group, which was extremely and significantly different from that in the blank control group (P < 0.001).CONCLUSION: Chondroitin sulfate with a proper concentration can accelerate the proliferation of HL60 cells, and it can increase the sensibility of HL60 cells to adriamycin, and promote the differentiation of HL-60 cells.
2.Velocity vector imaging evaluation of abdominal aorta atherosclerosis wall motion in rats
Danjun WANG ; Feng WANG ; Yun CHEN ; Shukun LU ; Yinting LIANG ; Yue WANG ; Yu SHI ; Zhengming HU
Chinese Journal of Medical Imaging Technology 2010;26(1):40-43
Objective To evaluate the characteristics of abdominal aorta wall motion in different stages of rats atherosclerosis with velocity vector imaging (VVI) technique. Methods Twenty-four healthy SD rats were on high-fat feeding after one week ordinary diet. Abdominal aortic intima-media thickness (IMT), end-systolic blood vessel diameter (Ds), peak systolic velocity (Vs), resistance index (RI), pulsatility index (PI) were measured before and at the end of 8th and 12nd week. Artery wall peak velocity (V_(max)), maximum tangential strain (S_(max)) and the maximum tangential strain rate (SR_(max)) were caculated with VVI. Results Abdominal aortic intima was rough and a small amount of foam cells were found under the light microscope at the end of 8 weeks of high-fat feeding. The values of Smax and SRmax measured at the end of 8th week of high-fat feeding decreased significantly than those of before high-fat feeding (P<0.05). At the end of 12nd week, abdominal aortic intimal was thicker and atherosclerotic plaque appeared somewhere. There were significant differences in artery IMT, Ds, Vs, RI, PI between before and the end of 2nd week of high-fat feeding (P<0.05);the values of V_(max), S_(max), SR_(max) decreased significantly than those of before and at the end of 8th week of high-fat feeding (P<0.05). Conclusion VVI can quantitatively evaluate the vessel wall elasticity in different stage of arteriosclerosis rats.
3.Effect and Mechanism of Steamed Allii Sativi Bulbus Polysaccharide Against Dysbiosis-induced Diarrhea in Young Rats
Yarong LI ; Mengyu GUO ; Yinting YU ; Nong ZHOU ; Weidong LI
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(13):77-84
ObjectiveTo verify the efficacy of steamed Allii Sativi Bulbus polysaccharide in regulating the intestinal flora of young dysbiosis-induced diarrhea rats based on 16S rRNA gene sequencing. MethodThe young SD rats were randomly divided into blank group,model group,positive drug group (bifid triple viable capsules),and high-dose and low-dose steamed Allii Sativi Bulbus polysaccharide groups,six in each group. The dysbiosis-induced diarrhea rat model was established,and the blank group and the model group were given normal saline by gavage (each 10 mL·kg-1),and the high-dose and low-dose steamed Allii Sativi Bulbus polysaccharide groups were administered with corresponding drugs (500 mg·kg-1 and 250 mg·kg-1, respectively) ,once a day for seven consecutive days. The loose stool rate,loose stool grade,diarrhea index,small intestine propulsion rate and hematoxylin-eosin (HE) staining were used as indexes to investigate the effect of steamed Allii Sativi Bulbus polysaccharide on improving diarrhea symptoms in young rats. The feces of rats were collected for 16S rRNA gene high-throughput sequencing. ResultCompared with the model group, the positive drug group and the high-dose and low-dose steamed Allii Sativi Bulbus polysaccharide groups had alleviated symptoms, down-regulated loose stool rate and diarrhea index (P<0.01) and decreased small intestine advancement rate (P<0.05). HE staining showed that after the treatment with steamed Allii Sativi Bulbus polysaccharide,the inflammatory cell infiltration of the colon tissue was improved and the intestinal gland recovered to the normal condition,which indicated that steamed Allii Sativi Bulbus polysaccharide could significantly ameliorate the diarrhea in young rats. The sequencing results revealed that steamed Allii Sativi Bulbus polysaccharide had a moderating effect on the abundance of the intestinal flora of young dysbiosis-induced diarrhea rats,elevating the flora richness and diversity indexes. Specifically, the abundance of Bacteroidota was increased while that of Firmicutes and Proteobacteria was decreased. ConclusionSteamed Allii Sativi Bulbus polysaccharide can be used to treat dysbiosis-induced diarrhea in young rats by regulating the abundance of intestinal microbiota.