1.Effect of lipopolysaccharide on mRNA expression of Toll-like receptor 4 and CD14 in lens epithelial cells
Xiurong HUANG ; Mingxin QI ; Jun WANG
Chinese Journal of Pathophysiology 2010;26(4):797-801
AIM: To investigate the effect of lipopolysaccharide (LPS) on the mRNA expression of Toll-like receptor 4 (TLR4) and CD14 in lens epithelial cells (LECs).METHODS: The cultured LECs were incubated with defferent concentrations of LPS for defferent times.The mRNA expression of TLR4 and CD14 in LECs were detected by reverse transcription polymerase chain reaction (RT-PCR).RESULTS: The mRNA expression of TLR4 in the LECs treated with LPS at concentrations of 50 μg/L, 100 μg/L, 200 μg/L, 500 μg/L or 1 000 μg/L was higher than that in the LECs without LPS treatment (P<0.01), respectively.The mRNA expression of TLR4 in the LECs incubated with 100 μg/L LPS for 24 h, 48 h or 72 h was higher than that in LECs without LPS treatment at same time points (P<0.01).The mRNA expression of CD14 in the LECs incubated with LPS at concentration of 100 μg/L for 24 h was also higher than that in the LECs without LPS treatment (P<0.01).CONCLUSION: LPS enhances the mRNA expression of TLR4 and CD14 in LECs, which may be related to intraocular response, as well as to the formation and development of after-cataract.
2.Apoptosis of lens epithelial cells induced by elemene
Xiurong HUANG ; Mingxin QI ; Keren KANG
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To investigate the effect of natural medicinal monomer elemene (Ele) on apoptosis of lens epithelial cells (LEC) and its the cellular and molecular mechanism in vitro. METHODS: The bovine LEC were cultured with Ele and used to observe the ultrastructure changes under transmission electron microscope and to detect DNA content and mitochondrial transmenbrane potential (??m) changes by flow cytometry. RESULTS: The typical morphological changes of LEC apoptosis in Ele group were observed under transmission electron microscope, such as chromatin condensation and aggregation at the nuclear periphery, and nuclear fragmentation as well. The DNA content of LEC in Ele group decreased and showed time-dependent. It was significant lower than that in control group (P
3.Recent advances in molecular mechanisms of oncosis
Yi CHEN ; Xiurong HUANG ; Mingxin QI
Chinese Journal of Pathophysiology 2000;0(11):-
Oncosis is another form of cell death, which is different from apoptosis. The review will discuss the recent advances of oncosis on pathological morphology, nuclear biochemical changes and molecular mechanisms. [
4.Advances in apoptosis induced by natural drugs
Keren KANG ; Xiurong HUANG ; Mingxin QI
Chinese Journal of Pathophysiology 1989;0(06):-
Apoptosis,a form of cell death,has generated considerable interest in recent years.Much progress has been made on the apoptosis induced by natural drgus,including component or mixture of plant,animal,mineral or marine materials,This review discusses some of the natural drugs that induce apoptosis and the possible mechanisms.
5.Inhibition effects of compound leech eye drops on apoptosis of lens epithelial cells and expressions of Bcl-2 and Bax genes in rats
Xiurong HUANG ; Mingxin QI ; Zhaoyang WANG ; Yi CHEN
Journal of Integrative Medicine 2007;5(6):681-5
OBJECTIVE: To investigate the inhibition effects of compound leech eye drops (Co-SZ) on apoptosis of lens epithelial cells (LECs) induced by hydrogen peroxide (H2O2) and the expressions of apoptosis-related genes Bcl-2 and Bax in rats. METHODS: All fresh transparent LECs in SD rats were bathed in culture medium with H2O2 in vitro, meanwhile Co-SZ were added in the culture medium. All LECs were incubated for 24 hours. The apoptosis rate of LECs was determined by terminal deoxynucleotidyl transferase mediated biotin-dUTP nick end labeling method (TUNEL). The changes of LEC ultrastructure and the formation of apoptotic body were observed by transmission electron microscopy. The expressions of apoptosis-related genes Bcl-2 and Bax were detected by streptavdin-peroxidase-biotin method. RESULTS: The apoptosis rate of LECs in the Co-SZ-treated group was significantly lower than that in the H2O2-treated group. The changes of apoptotic LEC ultrastructure in the Co-SZ-treated group were less than those in the H2O2-treated group. The expression of Bcl-2 protein was up-regulated and the expression of Bax protein was down-regulated in the Co-SZ-treated group as compared with the H2O2-treated group. CONCLUSION: The LEC apoptosis induced by H2O2 can be inhibited by Co-SZ. The molecular mechanisms of Co-SZ in inhibiting LEC apoptosis may be related to regulating the expressions of apoptosis-related genes Bcl-2 and Bax.
7.Effect of sodium ferulate on expression of bcl-2 and bax in rat lens epithelial cells
Mingxin QI ; Xiurong HUANG ; Zhaoyang WANG ; Yon WANG
Chinese Journal of Pathophysiology 2000;0(11):-
AIM: To investigate the effect of sodium ferulate on expression of apoptosis-related genes, bcl-2 and bax , in rat lens epithelial cells (LEC) injured by oxidation.METHODS: Eyes of SD rats were divided randomly into four groups: control group, hydrogen peroxide group (H 2O 2), pirenoxine sodium group (PS) and sodium ferulate group (SF). Eyes were excised and lenses were separated under operating microscope and sterilized condition. Lenses were incubated in CO 2 incubator for 24 h with 300 ?mol?L -1 H 2O 2 and with or rithout 5 mmol?L -1 SF. The expression of Bcl-2 and Bax protein of LEC were measured and compared by tearing the LEC anterior capsule via immunohistochemical analysis.RESULTS: (1) There were Bcl-2 and Bax expression in normal lenses of SD rates, Bcl-2 expression was stronger than Bax. (2) Bcl-2 expression decreased and Bax expression increased markedly ( P
8.Effects of schisandrin B on apoptosis of lens epithelial cells treated with H_2O_2
Xiurong HUANG ; Mingxin QI ; Zhaoyang WANG ; Yon WANG
Chinese Journal of Pathophysiology 2000;0(12):-
AIM: To investigate the effects of Schisandrin B (Sch B) on apoptosis of lens epithelial cells (LEC) treated with H 2O 2. METHODS: Eyes in SD rats were excised and lenses were separated under operating microscope in sterilized condition. Lenses were divided randomly into four groups with different treatment: control group, hydrogen peroxide group (H 2O 2), pirenoxine sodium group (PS) and schisandrin B group (Sch B). Lenses were incubated in CO 2 incubator for 24 h with 300 ?mol?L -1 H 2O 2 and with or without 0 5 mmol?L -1 Sch B. LEC aoptosis and apoptosis rate were measured by TUNEL method. Ultrastructure changes and apoptosis bodies of LEC were observed via transmitted electron microscope. RESULTS: (1) Apoptosis rate in H 2O 2 group (92.0?2.6) was significantly higher than that in control group (3.5?1.8). Apoptosis rate in Sch B group (13.8?3.27) was remarkably lower than that in H 2O 2 group and PS group. (2) Ultrastructure observation indicated that apoptosis cells occurred in most LEC in H 2O 2 group and the changes were severe presenting different stages. While a few apoptosis cells were observed in Sch B group, the changes were slight and most of them were in early and middle stages. CONCLUSION: These data indicated that Sch B significantly inhibited apoptosis of LEC during experimental oxidative injury, the effects were stronger than PS.
9.Inhibitory effects of Co-SZ eye drops on apoptosis of lens epithelial cells and their signal transduction mechanisms
Xiurong HUANG ; Mingxin QI ; Yi CHEN ; Chaoyang WANG ; Kunpeng LI
Chinese Journal of Pathophysiology 2000;0(10):-
AIM: To investigate the inhibitory effect of Co-SZ eye drops on apoptosis of lens epithelial cells(LEC) induced by H_2O_2 and to study the cellular and molecular mechanisms.METHODS:(1) All lenses of Sprague-Dawley rats were incubated with H_2O_2 and Co-SZ eye drops.The apoptosis rates of LEC were determined by TUNEL method.The changes of LEC ultrastructure and the formation of apoptotic body were observed by electron microscopy.(2) Bovine LEC were incubated with H_2O_2 and Co-SZ eye drops.The inhibitory of LEC apoptosis was detected by MTT after incubation.The changes of fractional DNA content in LEC were detected by flow cytometry(FCM).[Ca~(2+)]i,cAMP and cGMP of LEC were determined by spectrofluoremeter and radioimmunoassay,respectively.RESULTS: The LEC apoptosis rates in Co-SZ eye drops group were decreased significantly compared with H_2O_2 group by TUNEL.The ultrastructure changes in LEC of Co-SZ eye drops group were lighter than that in H_2O_2 group.The LEC apoptosis rates of Co-SZ eye drops group were dose-dependently decreased significantly compared with H_2O_2 groups via MTT assay.LEC apoptosis induced by H_2O_2 was inhibited by Co-SZ eye drops,and showing dose-dependent.The DNA contents in LEC of Co-SZ group were increased.The [Ca~(2+)]i and cAMP in Co-SZ group were decreased obviously.The cGMP was increased.CONCLUSION: The LEC apoptosis induced by H_2O_2 was inhibited by Co-SZ eye drops.The mechanism of apoptosis inhibition by Co-SZ eye drops maybe contribute to the increase in DNA content.The signal transduction mechanisms are related to the decrease in [Ca~(2+)]i and cAMP and the increase in cGMP.
10.Study on the mechanism of proliferative inhibition of arsenic trioxide and rhizoma curcumae on lens epithelial cell by MTT assay
Zhengzheng WU ; Xiurong HUANG ; Mingxin QI ; Jing YAN ; Yonhong HU
Ophthalmology in China 1993;0(01):-
Objective To investigate the proliferative inhibition of lens epithelial cell (LEC) by arsenic trioxide (ATO) and rhizoma curcumae (RC), in order to provide scientific basis for pursuing safe and effective natural drugs to prevent and cure after cataract. Design Experimental study. Participants Cultured Bovine LEC in vitro. Methods Changes of cellular form were observed with microscope. Different concentration of ATO (2.5, 5, 10?mol/L) and RC(5, 10, 20mg/ml) were added to the proliferative LEC separately. The effects of proliferative inhibition by ATO and RC on the LEC were measured with methyl thia-zolyl tetrazolium (MTT) assay method after 72 hours incubation. Main Outcome Measures Cellular form, Absorbance. Results Under the microscope, good growth and quantity increase were found in proliferative control group. Slowing proliferation,poor growth, and few disintegration were observed in ATO group and RC group. MTT assay: Different concentration of ATO (2.5, 5, 10?mol/L)and RC(5, 10, 20mg/ml) can significantly inhibit the proliferation of LEC and these effects were dose dependent(P