ABSTRACT
Objective To understand the effect of hyaluronic acid (HA) on the proliferation and apoptosis of chondrocytes cultured in vitro with Kashin-Beck disease(KBD) to provide the experimental evidences for treating KBD diseases with HA. Methods The articular cartilage samples collected from KBD patients were selected according to Diagnosis for Kaschin-Beck Disease(GB 16003-1995). And the normal cartilage samples were collected from victims of incidence (control). Chandrocytes were separated and cultured in vitro. Then varying dosages of HA were administered to chondrocytes and individed into 0,100,500 mg/L group, according to HA doages. The effect of HA on the proliferation and apoptosis of chondrocytes cultured/n vitro both KBD and the controls were investigated by methyl thiazolyl tetrazolium(MTT), Annexin V/PI staining on 2~(nd), 4~(th), 6~(th) day. Results In the control group, 500 mg/L group(0.140 ± 0.049) promoted chondrocyte proliferation significantly than 0 mg/L group (0.116 ± 0.021 ) at the 4~(th) day(P < 0.05), similar phenomenon was observed in KBD group in the 6~(th) day between 500 and 0 mg/L group(0.179 ± 0.081,0.128 ± 0.017, P< 0.05). In the KBD group, compared with 0 mg/L (12.860 ± 2.159), both 100 and 500 mg/L( 10.458 ± 1.143,7.877 ± 1.346) inhibited chondrocyte apoptosis rate (P < 0.05). In control, apoptosis rate of 500 mg/L group(4.045 ± 1.204) descreased compared with 0 mg/L group (7.128 ± 1.244, P < 0.05). Conclusion HA can promote the proliferation and inhibit the apoptosis of KBD chondrocytes cultured in vitro, and 500 mg/L HA play more effective role than that of 100 mg/L in promoting proliferation and inhibiting poptosis.
ABSTRACT
Objective To investigate the effect of selenium on proliferation and apoptosis of chondrocytes of articular cartilage cultured in vitro in Kaschin-Beck disease(KBD) patients and normal person, to explore the role of selenium in control of KBD, and to provide evidence for selenium's effect on the growth of normal cartilage cells. Methods The articular cartilage samples of grade Ⅱ and Ⅲ KBD patients were selected according to the national "Clinical Diagnosis of KBD" (GB 16003-1995). Chondrocytes of 5 KBD and 5 non-endemic normal accidentswere separated and cultured in vitro. KBD group and control group were given different doses of selenium (0,0.0125,0.0250,0.0500,0.1000,0.2500,0.5000,1.0000 mg/L, respectively). Methyl thiazolyl tetrazolium (MTT),flow cytometric analysis, and immunocytochemical staining were used to observe the effect of selenium on cell growth and apoptosis in KBD and normal persons. Results MTT results showed that the cell proliferation rate in each dosage group of the control group at the 6th day(0.086 ± 0.025,0.077 ± 0.012,0.073 ± 0.027,0.071 ± 0.017,0.058 ± 0.028,0.052 ± 0.028 and 0.046 ± 0.037) was significantly lower than that of 0 mg/L group(0.138 ± 0.026,all P ＜ 0.05);the average cell proliferation rate was negative( - 0.001 ± 0.001, - 0.003 ± 0.000, - 0.003 ± 0.001and - 0.004 ± 0.001 ) in 0.1000 - 1.0000 mg/L dose group, which was significantly lower than that of the 0 mg/L group(0.025 ± 0.003, all P ＜ 0.05);compared with 0 mg/L group(0. 115 ± 0.011), the KBD 0.2500 mg/L dose group promoted cell proliferation(0.128 ± 0.037, P ＜ 0.05), the KBD 1.0000 mg/L dose group inhibited cell growth (0.071 ± 0.019, P ＜ 0.05). The apoptotic rate of 0.0500 - 1.0000 mg/L dose control group [ (18.88 ± 0.02)%,(17.58 ± 0.01)%, (17.09 ± 0.04)%, (56.00 ± 0.02)%, (57.85 ± 0.03)% ] were higher than that of the 0 mg/L group[(13.51 ± 0.01)%, all P ＜ 0.05];compared with 0 mg/L group[(25.84 ± 0.02)%], the apoptotic rate in KBD 0.0250 - 0.2500 mg/L dose group [ ( 13.69 ± 0.02) %, ( 15.96 ± 0.03 ) %, ( 16.68 ± 0.03 ) %, ( 16.67 ± 0.02) % ]were lower, and the apoptotic rate in 0.5000, 1.0000 mg/L dose group [ (59.58 ± 0.03)%, (73.48 ± 0.04)% ] were significantly higher(all P ＜ 0.05). The Fas expression in KBD 0.0500 - 0.2500 mg/L dose groups[ (41.2 ± 1.5)%,(40.3 ± 2.0)%, (50.2 ± 2.5)%] were lower than those of the same dose control group with selenium intervention [(52.4 ± 1.0)%, (67.2 ± 4.0)%, (75.1 ± 5.0)%, all P ＜ 0.05], the caspase-3 expression in KBD 0.0500,0.1000 mg/L dose groups[ (40.8 ± 1.1 )%, (45.1 ± 2.1 )%] were lower than those of the same dose control group with selenium intervention[ (68.0 ± 3.0)%, (70.6 ± 3.5)%, all P ＜ 0.05 ]. Conclusions Appropriate dose of selenium supplementation (0.1000 - 0.2500 mg/L) could promote the growth of KBD chondrocyte, decrease cell apoptosis,but have a damage when the dose of selenium ＞ 0.5000 mg/L;doses of selenium that could promote the growth of KBD chondrocyte does not mean to promote the growth of normal cartilage cells in vivo.