1.Molecular mechanism of etodolac-induced apoptosis in SMMC7721 cell line
Xile ZHOU ; Jianjiang LIN ; Xiao XU ; Haiyang XIE ; Shusen ZHENG
Chinese Journal of Pathophysiology 1999;0(09):-
AIM: To investigate the possible role of nuclear transcription factor kappa B (NF-?B), Bcl-2, Bax and caspase-3 in etodolac-induced apoptosis of liver tumor SMMC7721 cell line. METHODS: Cell apoptosis was determined by flow cytometry analysis with PI staining and DNA laddering. Expression of Bcl-2 and Bax protein was measured by Western blotting. Caspase-3 activity was evaluated by active caspase-3 apoptosis kit with flow cytometry. NF-?B activation was detected by ELISA-based TransAM~(TM) NF-?B p65/p50 kit. RESULTS: Etodolac, a selective COX-2 inhibitor, stimulated apoptosis in liver tumor SMMC7721 cell line significantly. Flow cytometry showed that the apoptotic rate was 16.3%?3.1%, 19.9%?3.6%, 22.9%?3.2%, 31.2%?3.3% with different concentrations of etodolac (0.25, 0.50, 1.0 or 2.0 mmol/L), while the apoptotic peak did not appear in the control group (0 mmol/L) (P
2.The expression of BMP-2 mRNA of rat osteoblast cell sheets cultured with ascorbic acid
Chengju ZHOU ; Jianjiang ZHOU ; Hong YANG ; Jiufeng MAO ; Xiao ZENG ; Yuanyuan JIA ; Yuting WANG ; Qiang DONG
Journal of Practical Stomatology 2015;(3):305-308
Objective:To study the effects of ascorbic acid on BMP-2 mRNA expression of osteoblast cell sheets.Methods:Rat os-teoblasts were primaryly cultured and identified;osteoblast cell sheets were built by physical scraping method in vitro;the osteoblast cell sheets were cultured with 1 5,50 and 85 mg/L ascorbic acid for 1 and 2 weeks respectively,and the expression of BMP-2 mRNA of the cell sheets was detected by RT-qPCR.Results:The obtained cells were conformed to be osteoblasts.The osteoblast cell sheets could be rolled into tube in vitro.The expression of BMP-2 mRNA of osteoblast cell sheets in experiment group,whether in week one or week two was higher than that in control group,50 mg/L group showed the highest expression(first week P <0.05;second week P>0.05);the expression of any group in week two was higher than that in week one(P <0.05).Conclusion:Ascorbic acid may pro-mote the expression of BMP-2 mRNA in osteoblast cell sheets.
3.Resection of tracheal tumor under cardiopulmonary bypass: a case report.
Hai-hong CHEN ; Wei PAN ; Xiao-xia AN
Chinese Medical Journal 2005;118(12):1047-1049
4.The effects of Curcumin on the cellular apoptosis induced by high glucose in rat retinal vascular endothelial cells
Jiang HUANG ; Yi LI ; Jianjiang XIAO ; Guoxu XU ; Shuyang BU ; Weifeng LUO
Chinese Journal of Ocular Fundus Diseases 2017;33(5):513-517
Objective To observe the effects of Curcumin on the cellular apoptosis of rat retinal vascular endothelial cells (RRVEC) induced by high glucose. Methods Generation 4 cultured RRVEC were used in this experiment, and identified with anti-vWF factor antibody by immunochemistry and immunofluorescence. The RRVEC were divided into control group (5.5 mmol/L glucose), high glucose group (30 mmol/L glucose), and treatment group (30 mmol/L glucose+30 μmol/L Curcumin), respectively. Flow cytometry was used to measure the cellular reactive oxygen species (ROS) level and apoptosis. The expression intensity and location of nuclear factor (NF)-κB p65 in the cells of the three groups were detected by immunochemistory. The expression of Bcl-2 and Bax protein was detected by Western blot test. Results Immunostaining showed that RRVEC were positive for vWF factor. The flow cytometry showed that the cellular ROS level in treatment group was higher than that in the control group (t=8.677, P=0.000), but less than that in the high glucose group (t=40.957, P=0.000). Compared with the high glucose group, the cellular ROS level in the treatment group was decreased significantly (t=6.568, P=0.000). The cellular apoptosis were significantly different among the three groups (F=325.137, P=0.000). Compared with the high glucose group, the cellular apoptosis in the treatment group was decreased significantly (t=12.818, P=0.000). Immunochemistry showed that NF-κB p65 was expressed strongly in the cellular nuclei and cytoplasm in the high glucose group than that in the control group and the treatment group with the significant differences (t=8.322, P=0.000). Western blot results demonstrated that compared with the control group, the expression of Bcl-2 of RRVEC and Bcl-2/Bax ratio decreased (t=4.362, 6.449; P=0.005, 0.001) and Bax increased (t=3.813, P=0.009)in the high glucose group, with statistically significant differences. Compared with the high glucose group, the expression of NF-κB and Bax decreased (t=2.577, 3.059; P=0.042, 0.022) and Bcl-2/Bax ratio increased significantly (t=3.831, P=0.009) in the treatment group. Conclusion Curcumin could suppress the cellular apoptosis of RRVEC induced by high glucose. The mechanism of Curcumin protecting RRVEC may be via regulating NF-κB signal pathway.