- 學位論文
探討維生素C、礦物元素硒及硫化軟骨素對緩解骨關節炎之效用與機制
Study on the Effects and Mechanisms of Vitamin C, Selenium and Chondroitin Sulfate on Osteoarthritis
摘要
骨關節炎（Osteoarthritis）為一與老化相關之疾病。關節在經年累月使用之下，關節軟骨之細胞外基質會逐漸減少，使軟骨細胞失去基質包覆，而軟骨細胞因缺乏外在保護及活性物質供給等諸多因素，導致軟骨細胞死亡，最終使軟骨分解。維生素C及微量礦物元素硒為維持人體健康所必需之營養物質，但其對保護軟骨細胞是否具效用仍不明。已知，碘醋酸鈉（mono - iodoacetate，MIA）可誘發骨關節炎。本研究透過細胞實驗（in vitro）探討維生素C及硒對人類軟骨瘤細胞SW-1353受到MIA傷害時之保護機制。此外，亦透過動物實驗（in vivo）來探討市售之骨關節保健食品硫化軟骨素（chondroitin sulfate sodium salt，CS）其緩解以MIA誘發大鼠骨關節炎之能力。 骨關節炎中軟骨細胞死亡常因受到活性氧物質（reactive oxygen species，ROS）的攻擊而致使軟骨分解的重要原因。其中Nrf2 （Nuclear factor-erythroid 2-related factor 2）為受氧化壓力調控並與細胞內多種抗氧化酵素表現相關的轉錄因子。研究發現，經維生素C及硒處理之細胞，可有效防止MIA抑制Nrf2的轉錄作用，以提供抗氧化能力進而防止MIA之傷害。而在細胞凋亡方面，在維生素C及硒的處理之下，可有效降低因MIA作用而使細胞停留在sub-G1，且可透過活化轉錄因子NF-κB及p53進而抑制細胞凋亡相關蛋白如Bax及cytochrome c的表現。同時也抑制了pro-caspase 9及pro-caspase 3的活化，避免軟骨細胞走向細胞凋亡。此外，當DNA受到損傷時，體內為了維持基因的穩定性，而發展出可隨時監控損害發生並即時活化DNA修復的訊息傳遞路徑。研究發現，以維生素C及硒處理之細胞，其DNA修復相關蛋白：Ataxia telangiectasia mutated（ATM）及Nibrin（NBS1）表現量回升，使細胞凋亡情形被抑制。而在MIA誘導之大鼠骨關節炎方面，於病理組織染色結果顯示，硫化軟骨素能有效維持軟骨細胞之生理活性及降低軟骨基質磨損，以延緩骨關節炎病程的進行。 綜合上述，維生素C及微量礦物元素硒在細胞實驗被證實具有保護軟骨細胞傷害之功效。在大鼠動物骨關節炎模式上，亦發現硫化軟骨素可減弱骨關節炎之惡化程度。
並列摘要
Osteoarthritis（OA） is an age-related degenerative joint disease which characterizes by the loss of cartilage extracellular matrix and results in chondrocytes’ death and cartilage damage. Vitamin C and selenium (Se), a trace mineral element, are essential nutrients for people to maintain health, however, it is still not known whether they are protective against OA or not. The mono-iodoacetate（MIA）has been known to induce OA in rats. In this study, we designed a series of experiments to figure out the effects and mechanisms of vitamin C and Se on the protection against the MIA-damaged SW-1353 cells. In addition, we also explore chondroitin sulfate sodium salt (CS), a health food, on improving abrasion of articular cartilage. Over-production of reactive oxygen species (ROS) is one of the major causes of OA; thus, increase in anti-oxidative capacity to remove ROS could lessen the severity of this disease. Transcription factor of nuclear factor-erythroid 2-related factor 2（Nrf2） is responsible for regulating a variety of antioxidant enzymes’ expressions. Our results showed vitamin C and Se could increase Nrf2 expression which was decreased in the MIA-treated SW-1353 cells. Accordingly, we proposed that the anti-oxidative capacity of MIA-treated SW-1353 cells might be enhanced after vitamin C and Se additions. Moreover, vitamin C and Se additions could decrease the MIA-induced SW-1353 cell’s apoptosis by activating NF-κB and p53. The expressions of apoptotic proteins, such as Bax and cytochrome c, and the activations of pro-caspase 3 and pro-caspase 9 were reduced. In order to maintain genomic stability, there are several DNA repair systems are activated as DNA is damaged. Interestingly, we also found the expressions of DNA repair-related proteins including Ataxia telangiectasia mutated (ATM) and Nibrin (NBS1) were also increased after vitamin C and Se treatment. In a MIA-induced OA model of rat, the pathological results showed CS would lessen the stress of chondrocytes and improve abrasion of articular cartilage. In Conclusion, This study demonstrates that either vitamin C, Se or CS is protective against OA progression. It seems worthwhile to further evaluate their clinical efficacy.
參考文獻
延伸閱讀
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