茶多酚对饮茶型氟中毒大鼠关节软骨氧化损伤的保护作用
Protective role of tea polyphenols in oxidative stress damage of the rat articular cartilage tissue caused by brick-tea fluorosis
摘要目的 探讨茶多酚对饮茶型氟中毒大鼠关节软骨的氧化应激损伤的保护作用.方法 120只雄性Wistar大鼠按体质量随机分为6组:对照组、加氟组、氟+茶多酚组、氟+铝组、氟+铝+茶多酚组和砖茶组.加氟组每日饮用含氟(F-)100.00 mg/L的氟化钠(NaF)水溶液;氟+茶多酚组每日饮用含F-100 mg/L、茶多酚10.0 g/L的水溶液;氟+铝组每日饮用含F-100.00 mg/L、铝(Al3+)200.00 mg/L的水溶液;氟+铝+茶多酚组同时饮用含有上述3种物质的水溶液:砖茶组饮用砖茶沏制而成的砖茶水(F- 100.00 mg/L、Al3+215.00mg/L、9.2 g/L);对照组饮用自来水(F- 0.33 mg/L).连续饲养3个月,处死动物,检测血清中超氧化物歧化酶(SOD)、总抗氧化能力(T-AOC)、戊二醛(MDA)、一氧化氮(NO)和细胞因子白介素1β(IL-1β)、白介素6(IL-6)的水平;RT-PCR和免疫组化法检测关节软骨中诱生型一氧化氮合酶(iNOS)mRNA及蛋白表达.结果 氟 +铝+茶多酚组SOD水平[(664.009±29.589)kU/L]与加氟组、氟+销组[(625.328±27.199)、(652.282±13.926)kU/L]比较有升高趋势,但是差异无统计学意义(P均>0.05);氟+茶多酚组、氟+铝+茶多酚组、砖茶组T-AOC水平[(10.874±0.721)、(11.871±0.941)、(10.380±2.747)kU/L]与加氟组、氟+铝组[(8.849±1.887)(8.210±1.740)kU/L]比较,差异有统计学意义(P均<0.05);氟+铝+茶多酚组血清中MDA水平[(3.235±0.446)μmol/L]与加氟组、氟+铝组[(3.889±0.387)、(4.580±0.474)μmol/L]比较,差异有统计学意义(P均<0.05);氟+茶多酚组、氟+铝+茶多酚组、砖茶组血清中NO水平[(23.278±2.386),(20.643±2.623)、(24.367±6.072)μmol/L]与加氟组、氟+铝组[(32μ962±8.268)、(34.909±6.288)μmol/L]比较,差异有统计学意义(P均<0.05):加氟组、氟+销组、氟+茶多酚组、氟+铝+茶多酚组、砖茶组血清IL-1β水平分别为(4.728±0.297)、(4.412±0.229)、(4.432±0.285)、(4.516±0.351)、(4.614±0.2270)ng/L,组间比较,差异无统计学意义(F=2.314,P>0.05);氟+铝+茶多酚组、砖茶组IL-6水平[(7.231±0.596)、(7.325±0.290)ng/L]与氟+铝组[(8.256±0.635)ng/L]比较,差异有统计学意义(P均<0.05).氟+茶多酚组、氟+铝+茶多酚组、砖茶组iNOS mRNA相对表达量(0.482±0.021、0.447±0.021、0.491±0.022)与加氟组、氟+铝组(0.562±0.025、0.591±0.020)比较,差异有统计学意义(P均<0.05);对照组iNOS蛋白表达阳性细胞主要分布在关节表层,各实验组iNOS阳性细胞在关节面表层、中层均有分布.结论 茶多酚能通过清除氧自由基、提高机体总抗氧化能力、减少脂质过氧化产物等抗氧化作用减轻饮茶型氟中毒引起的大鼠氧化应激损伤,对饮茶型氟中毒有一定的保护作用.
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abstractsObjective To explore the protective mechanism of tea polyphenols (TPs) ion oxidative stress damage of the rat articular cartilage tissue caused by brick-tea fluorosis. Methods One hundred and twenty wistar male rats were randomly divided into 6 groups according to body mass: fluoride group with drinking water containing 100.00 mg/L F-, fluoride plus TPs group treated with 100.00 mg/L F- and 10.0 g/L TPs, fluoride plus aluminum group fed with 100.00 mg/L F- and 200.00 mg/L Al3+, fluoride plus aluminium and TPs group treated with 100.00 mg/L F-,200.O0 mg/L Al3+ and 10.0 g/L TPs;brick-tea group treated with drinking water containing 100.00 mg/L F-,215.00 mg/L Al3+ and 9.2 g/L TPs, which was steeped by the brick-tea;control group treated with tap water. The animals were bred for three months and then sacrificed. The level of SOD,T-AOC and MDA in blood serum were detected,also the level of NO and cytokine IL-1β and IL-6, the expression of iNOS mRNA and protein in articular cartilage were respectively analyzed by RT-PCR and immunohistochemistry. Results Blood serum SOD level in the fluoride plus aluminum and TPs group[(664.009 ± 29.589)kU/L] was higher compared with that in the fluoride group[(625.328 ± 27.199)kU/L], fluoride plus aluminum group[(652.282±13.926)kU/L], although no statistically significant differences was found(P > 0.05) ;blood serum T-AOC level of the fluoride plus TPs, fluoride plus aluminum and TPs group, brick tea group[(10.874 ± 0.721), (11.871 ± 0.941), (10.380 ± 2.747)kU/L] was higher compared with fluoride group, fluoride plus aluminum group [(8.849 ± 1.887), (8.210 ± 1.740)kU/L], the differences all being statistically significant(P < 0.05) ;blood serum MDA level in the fluoride plus aluminum and TPs group[(3.235 ± 0.446)μmol/L] had significances compared with fluoride group, fluoride plus aluminum group [(3.889 ± 0.387), (4.580 ± 0.474)μmol/L, all P < 0.05)];blood serum NO level in fluoride plus Tps group, fluoride plus aluminum and TPs group, brick-tea group[(23.278 ± 2.386), (20.643 ± 2.623), (24.367 ± 6.072) μmol/L] had tatistical differences compared with fluoride group, fluoride plus aluminum group[(32.962 ± 8.268), (34.909 ± 6.288)μmol/L, all P < 0.05];blood serum IL-1β level of fluoride group, fluoride plus aluminum, fluoride plus Tps, fluoride plus aluminum and TPs group and brick-tea group [(4.728 ± 0.297), (4.412 ± 0.229), (4.432 ± 0.285), (4.516 ± 0.351), (4.614 ±0.2270)n/L] did not have inter-group differences (F = 2.314,P > 0.05);the blood serum IL-6 level of fluoride plus aluminum and TPs group, brick-tea group[(7.231 ± 0.596), (7.325 ± 0.290)ng/L] had statistical differences compared with fluoride plus aluminum[(8.256 ± 0.635)ng/L, P < 0.05]. The iNOS mRNA correspondent expression content of fluoride plus Tps group, fluoride plus aluminum and TPs group, brick-tea group(0.482 ± 0.021,0.447±0.021,0.491 ± 0.022) had statistical differences compared with fluoride group, fluoride plus aluminum group (0.562 ± 0.025,0.591 ± 0.020, all P < 0.05). Cells with positive iNOS protein expression of control group were mainly distributed at the surface layer of joint, while the cells of experiment groups were distributed both at the surface layer and the intermediate layer. Conclusions Tea polyphenols could alleviate oxidative stress damage on the articular cartilage, exerting protection against brick-tea fluorosis on rats through cleaning up free radicals, elevating total anti-oxidation capability, diminishing the generation of lipid peroxide.
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