CAJ | 학술논문

目的研究氟对大鼠切牙细胞Fas表达及胱天蛋白酶(caspase)3和caspase-8活性和细胞凋亡的影响,以期进一步探讨氟对牙齿的作用以及氟斑牙的发生机制.方法将40只SD大鼠按随机数字表法随机分为4组,每组10只.低剂量染氟组(低氟组)、中剂量染氟组(中氟组)和高剂量染氟组(高氟组)分别饮用以蒸馏水配制的含10、50和100 mg/L NaF的高氟水;对照组饮用不添加NaF的蒸馏水;在60、90 d时各组分别处死5只动物.用免疫组化法检测Fas的表达,采用酶标仪检测caspase-3和caspase-8的活性,用流式细胞术检测大鼠切牙细胞凋亡.结果在染氟60 d时,对照组、低氟组、中氟组及高氟组的Fas表达结果分别为:0.1819±0.0025、0.2120±0.0084、0.2283±0.0183及0.2818±0.0233;在染氟90 d时,对照组、低氟组、中氟组及高氟组的Fas表达结果分别为:0.2077±0.0289、0.2216±0.0105、0.2377±0.0059及0.2775±0.0088.大鼠切牙细胞Fas表达和caspase-3活力随染氟剂量增高而增强,存在显著的剂量-效应关系,实验60及90 d时Fas的相关系数分别为0.9728(P<0.01)和0.9889(P<0.01);caspase-3的相关系数分别为0.9533(P<0.01)和0.9849(P<0.01).在染氟90 d时,大鼠切牙细胞凋亡率和caspase-8活力随染氟剂量增高而增强,凋亡率和caspase-8的相关系数分别为0.9733(P<0.01)和0.9928(P<0.01).在染氟60和90 d时,大鼠切牙细胞Fas表达与caspase-3活力之间存在明显相关关系,相关系数分别为0.9619(P<0.01)和0.9912(P<0.01);在染氟90 d时,大鼠切牙细胞Fas表达与凋亡率和caspase-8活力之间也有明显的相关关系,Fas与凋亡率和caspase-8的相关系数分别为0.9841(P<0.01)和0.9767(P<0.01).结论在10、50和100 mg/L NaF剂量条件下染氟60和90 d,大鼠切牙细胞Fas表达增强并介导caspase激活和细胞凋亡,详尽机制仍需深入探讨.
목적 연구불대대서절아세포Fas표체급광천단백매(caspase)3화caspase-8활성화세포조망적영향,이기진일보탐토불대아치적작용이급불반아적발생궤제.방법 장40지SD대서안수궤수자표법수궤분위4조,매조10지.저제량염불조(저불조)、중제량염불조(중불조)화고제량염불조(고불조)분별음용이증류수배제적함10、50화100 mg/L NaF적고불수;대조조음용불첨가NaF적증류수;재60、90 d시각조분별처사5지동물.용면역조화법검측Fas적표체,채용매표의검측caspase-3화caspase-8적활성,용류식세포술검측대서절아세포조망.결과 재염불60 d시,대조조、저불조、중불조급고불조적Fas표체결과분별위:0.1819±0.0025、0.2120±0.0084、0.2283±0.0183급0.2818±0.0233;재염불90 d시,대조조、저불조、중불조급고불조적Fas표체결과분별위:0.2077±0.0289、0.2216±0.0105、0.2377±0.0059급0.2775±0.0088.대서절아세포Fas표체화caspase-3활력수염불제량증고이증강,존재현저적제량-효응관계,실험60급90 d시Fas적상관계수분별위0.9728(P<0.01)화0.9889(P<0.01);caspase-3적상관계수분별위0.9533(P<0.01)화0.9849(P<0.01).재염불90 d시,대서절아세포조망솔화caspase-8활력수염불제량증고이증강,조망솔화caspase-8적상관계수분별위0.9733(P<0.01)화0.9928(P<0.01).재염불60화90 d시,대서절아세포Fas표체여caspase-3활력지간존재명현상관관계,상관계수분별위0.9619(P<0.01)화0.9912(P<0.01);재염불90 d시,대서절아세포Fas표체여조망솔화caspase-8활력지간야유명현적상관관계,Fas여조망솔화caspase-8적상관계수분별위0.9841(P<0.01)화0.9767(P<0.01).결론 재10、50화100 mg/L NaF제량조건하염불60화90 d,대서절아세포Fas표체증강병개도caspase격활화세포조망,상진궤제잉수심입탐토.
Objective To investigate the effects of fluoride on Fas expression, caspase-3 and caspase-8 activity and apoptosis in rat incisor cells. Methods Forty male SD rats were divided into 4 groups randomly and provided with distilled water containing NaF at the doses of 0, 10, 50 and 100 mg/L respectively. Each group had 10 animals. Five animals were sacrificed at 60 and 90 days respectively. Fas expression was measured with immunohistochemistry, and colorimetric assay was used to examine caspase-3 and caspase-8 activity with enzyme-labelled meter. The apoptosis was detected by flow cytometry in mandibular incisor cells. Results NaF at the doses of 10,50 and 100 mg/L for 60 d and 90 d caused Fas overexpression, promoted activity of caspase-3 and caspase-8, increased apoptosis rate in mandibular incisor cells. At 60 days,the value of Fas expression was 0.1819±0.0025 for control, 0.2120±0.0084 for 10 mg/L NaF group, 0.2283±0.0183 for 50 mg/L NaF group, 0.2818±0.0233 for 100 mg/L NaF group. At 90 days,the value of Fas expression was 0.2077±0.0289 for control, 0.2216±0.0105 for 10 mg/L NaF group, 0.2377±0.0059 for 50 mg/L NaF group, 0.2775±0.0088 for 100 mg/L NaF group. Statistics analysis yielded close relationship between the dose of NaF in water and the Fas expression, and also between the dose of NaF in water and caspase-3 activities, and the relative coefficient was 0.9728(60 d, P<0.01) and 0.9889(90 d, P<0.01)for Fas expression, 0.9533(60 d, P<0.01)and 0.9849(90 d, P<0.01)for caspase-3 activity respectively. Apoptosis rate and caspase-8 activity also had close relationship with the NaF doses, and the relative coefficient was 0.9733(90 d,P<0.01)for apoptosis, 0.9928(90 d, P<0.01)for caspase-8. At the doses of 10, 50 and 100 mg/L NaF for 60 d and 90 d, obvious relationship was found between Fas expression and caspase-3 activity, and the relative coefficient was 0.9619(60 d, P<0.01) and 0.9912(90 d, P<0.01). Obvious relationship between Fas expression and apoptosis, between Fas expression and caspase-8 activity was found in groups for 90 d, and the relative coefficient was 0.9841(P<0.01)for apoptosis, 0.9767(P<0.01)for caspase-8. Conclusions Fluoride could induce Fas overexpression and mediate caspase activation and apoptosis at the doses of 10,50 and 100 mg/L for 60 d and 90 d in rat mandibular incisor cells.