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1.
International Journal of Pediatrics ; (6): 239-243, 2016.
Article in Chinese | WPRIM | ID: wpr-485347

ABSTRACT

Objective To establish and evaluate intestinal epithelial barrier model using Caco-2 cell so as to play a foundation for next study of barrier permeability.Methods Caco-2 cells were cultured in vitro then seeded into Transwell cell culture inserts.The permeability of the intestinal epithelial barrier was detected by transepithelial electrical resistance(TEER)and lucifer yellow flux,and verified by transmission electron micro-scope.Different concentrations of PAF(0,50,100,and 200 nmol /L)were exposed for 24 hours to Caco-2 mono-layer when cultured 21 days.The tight junction was observed under transmission electron microscope.Assess-ment of ZO-1 protein localization and expression were detected by immunofluorescence and Western blot analy-sis.Results Cultured Caco-2 cell confluencd as monolayer with time passed.From 5th day,TEER increased, then reached 600Ω?cm2 at 15th day and lasted to 21 st day,there was little flux of lucifer yellow,transmission e-lectron microscopy also found cells differentiated better,had well-arranged villi and polarity alined as monolayer, forming completed tight junction which was the marker of intestinal epithelial barrier model in vitro.TEER de-creased and lucifer yellow flux increased in cells exposed to PAF.The permeability reached the peak when ex-posed to 100 nmol /L PAF(P <0.01 ),tight junction disrupted,ZO-1 protein expression downregulated,abnor-mal localization and distribution was assessed by immunofluorescence staining.Conclusion Cultured Caco-2 cells for 2-3w can be used to study intestinal epithelial barrier as a model in vitro.PAF increased intestinal epi-thelial permeability,which would correlate to the decreased protein expression and abnormal distribution of ZO-1.

2.
Chinese Traditional Patent Medicine ; (12): 374-378, 2010.
Article in Chinese | WPRIM | ID: wpr-433338

ABSTRACT

AIM:To check the antiproliferative and apoptosis-inducing effect of Compound Changtai Granule(Radix et Rhizoma ginseng,Semen coicis,Rhizoma curcumae,etc)(CCG)on human colon carcinoma cell line(SW480).METHODS:Methy thiazolyl tetrazolium chromatomerry was used to observe the influence of several concentration of CCG on SW480 cell proliferation cultured in vitro at the various times,and flow cytometry was applied to examining the change in SW480 cell proliferation cycle and apoptosis rate.RESULTS:CCG could successfully prevent SW480 cell from entering G_0/G_1 phase and G_2/M phase,decrease cell numbers in S phase at the same time.CONCLUSION:CCG has an obvious dose-response relationship in the range of 3.51 to 7.81 mg/mL.

3.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 399-402, 2007.
Article in Chinese | WPRIM | ID: wpr-238739

ABSTRACT

To investigate the effect of meloxicam, a selected NSAIDs, on cell growth, expression of VEGF and angiopointin-2 (Ang-2) protein in HT-29 cell line, cultured HT-29 cells were treated with meloxicam of various concentrations for various lengths of time. The proliferation of HT-29 was detected by cell counting kit-8 (CCK-8), the cell cycle was determined by flow cytometer and the levels of VEGF and Ang-2 protein in supernatants were examined by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of VEGF and Ang-2 in cultured HT-29 were determined by real-time quantitative reverse-transcription polymerase chain reaction. Our results showed that treatment of meloxicam of different concentrations and for various lengths of time had a cytotoxicic effect on the cell proliferation of HT-29 cells in a concentration-dependant and time-dependant manner. Cell cycle analysis showed that the cells were mainly blocked in G0/G1 phase. The VEGF and Ang-2 protein levels in supematants of the culture medium were decreased gradually in a concentration-dependent or time-dependent fashion. The mRNA expression of cox-2, VEGF and Ang-2 showed a gradual and concentration-dependent reduction. It is concluded that meloxicam can reduce the expression of VEGF and Ang-2 at the protein and mRNA level in colon carcinoma cell line.

4.
Chinese Journal of Current Advances in General Surgery ; (4)1999.
Article in Chinese | WPRIM | ID: wpr-545263

ABSTRACT

Objective: To study the affection of Diallyl trisulfide on cultured human colon carcinoma cell line HT-29 and its mechanism. Methods: The viability of cells after various treatments were determined using the method of MTT. Flow cytometry were used to analyze the change of cell cycle and Bcl-2, Bax levels. Results: The proliferation of human colon carcinoma HT-29 cells were inhibited by Diallyl trisulfide. The inhibition was associated with dose and time dependence. Flow cytometry results showed that: 1) When cells were treated with Diallyl trisulfide at the concentration of 10 ?g/mL for 12 h and 24 h, the percentage of G0/G1 phase cells was decreased and that of G2/M phase cells was significantly increased compared with those in the control group.2) Diallyl trisulfide down regulated Bcl expression, while up-regulated bax expression. Conclusions: It was suggested that Diallyl trisulfide inhibited HT29 cell proliferation. And these were associated with arrest of cell cycles and down regulation of Bcl2/Bax ratio.

5.
Chinese Traditional Patent Medicine ; (12)1992.
Article in Chinese | WPRIM | ID: wpr-580710

ABSTRACT

AIM:To check the antiproliferative and apoptosis-inducing effect of Compound Changtai Granule(Radix et Rhizoma ginseng,Semen coicis,Rhizoma curcumae,etc)(CCG) on human colon carcinoma cell line(SW480).METHODS:Methy thiazolyl tetrazolium chromatomerry was used to observe the influence of several concentration of CCG on SW480 cell proliferation cultured in vitro at the various times,and flow cytometry was applied to examining the change in SW480 cell proliferation cycle and apoptosis rate.RESULTS:CCG could successfully prevent SW480 cell from entering G0/G1 phase and G2/M phase,decrease cell numbers in S phase at the same time.CONCLUSION:CCG has an obvious dose-response relationship in the range of 3.51 to 7.81 mg/mL.

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