凋亡細胞清除相關基因多型性與台灣地區全身性紅斑性狼瘡及類風濕性關節炎發生之探討

Investigating of Genetic Polymorphisms on Apoptotic Cells Clearance-Related Genes in Systemic Lupus Erythematosus and Rheumatoid Arthritis in Taiwanese Population

卓映君(Ying-Chun Cho)

指導教授 ： 胡忠怡

國立臺灣大學/醫學院/醫學檢驗暨生物技術學研究所/碩士(2010年)

https://doi.org/10.6342/NTU.2010.00360

摘要

人體中，每天都約有109 以上的細胞會進行細胞凋亡更替以維持身體的恆定性。當細胞發生細胞凋亡以後，會釋放出化學性趨化物質誘使吞噬細胞往凋亡細胞移動。吞噬細胞藉由辨識凋亡細胞上暴露出的“eat-me＂訊息分子後，能夠將凋亡細胞吞噬(efferocytosis)，並且釋放出抗發炎的細胞激素。若凋亡的細胞無法被即時的清除，就會發生細胞壞死(secondary necrosis)、造成組織處在慢性發炎的狀態，或導致自體免疫疾病的產生。在多個小鼠實驗模型皆証實凋亡細胞清除有缺陷足以造成類似人類狼瘡疾病。為了驗證凋亡細胞清除遲緩與人類自體免疫疾病發生是否相關，本研究探討近年來發現參與凋亡細胞清除相關分子: 包含Stabilin-2、Tim4 (T-cell immunoglobulin and mucin domains-containing protein 4)、Tim3、Tim1、Bai1 (Brain-Specific Angiogenesis Inhibitor1)、HMGB-1 (High mobility group box 1)基因變異性與台灣地區全身性紅斑性狼瘡或類風濕性關節炎罹病之關聯性。
本研究共收集台灣地區136 位全身性紅斑性狼瘡病患與143 位健康對照個案，以及141 位類風濕性關節炎病患和160 位健康對照個案。國人的Tim3、Tim4、HMGB-1 在造成胺基酸改變的基因多型性位點上並未發現變異，以序列特異性寡核苷酸探針雜交或限制酶片段長度多型性方式檢視比較受試檢體之STAB2、BAI1核苷酸多型性之基因型。結果顯示，STAB2 +1529 C/A 位點之A 對偶基因與全身性紅斑性狼瘡罹病為正相關(OR=2.2, 95 % CI=1.1~4.2, p=0.016)。STAB2 +5207 A/C 位點基因型分佈在全身性紅斑性狼瘡病患與對照組雖未達顯著差異(p=0.057)，但對偶基因C 攜帶率與全身性紅斑性狼瘡患者發生中樞神經系統病變有顯著正相關(OR=5.0, 95% CI=1.62~15.5, p=0.007)。STAB2 +1529 C/A、STAB2 +5207 A/C 位點與類風性關節成病並不相關。STAB2 [phone] C/G-G/T 單倍體出現率之分佈與全身性紅斑性狼瘡和類風濕性關節炎皆無關。BAI1 基因三端非轉譯區之+5010 G/C位點上C 對偶基因出現率在全身性紅斑性狼瘡及類風濕性關節炎病患呈顯著正相關(SLE: p=0.007, RA: p=0.019)。本研究發現若干參與清除凋亡細胞分子的多型性與國人罹患自體免疫疾病相關聯，暗示凋亡細胞清除遲緩，可能為自體免疫疾病成病原因之一。未來應再進一步檢視STAB2 +1529 C/A 、BAI1 +5010 G/C 位點變異代表的生物分子意義，以確定二者是否有潛力成為國人全身性紅斑性狼瘡或類風濕性關節炎基因標誌。

關鍵字

凋亡細胞清除 ； 全身性紅斑性狼瘡 ； 類風濕性關節炎 ； 核苷酸多型性 ； Stabilin-2 ； Bai1

並列摘要

More than 109 cells are undergoing apoptosis in human body daily in the process related to homeostasis maintenance. Apoptotic cells release chemoattractants to attract phagocytes. The recruited phagocytes recognize and engulf apoptotic cells through “eat me”signals presented on apoptotic cells. The so called “efferocytosis” process enhances release of anti-inflammatory cytokines. In case of deficiency in efferocytosis, apoptotic cells undergo secondary necrosis, which result in chronic inflammation in local tissues or auto-immunity. Evidences in several marine models supported that lacking apoptotic cell clearance leads to lupus-like disease. To clarify weather delayed apoptotic cell clearance could contribute to human autoimmune diseases, we intend to investigate the genetic polymorphisms of several efferocytosis related molecules reported in recent years, including Stabilin-2, Tim4 (T-cell immunoglobulin and mucin domains-containing protein 4), Tim1, Tim3, Bai1 (Brain-Specific Angiogenesis Inhibitor 1),and HMGB-1 (High mobility group box 1) and their correlation to systemic lupus erythematosus (SLE) or rheumatoid arthritis (RA) in Taiwan.
In this study, we studied 136 SLE patients with 143 age, gender-matched non-lupus , and 141 RA patients with 160 age, gender-matched control. There are rare non-synonymous genetic polymorphisms in Tim3, Tim4, and HMGB-1 found in Taiwanese population. The genetic polymorphisms of STAB2 and BAI1 in the studied subjects were examined by PCR/SSOPH (Sequence-specific oilgonucleotide probe hybridization) or PCR/RFLP (Restriction fragment length polymorphisms). We found that carriage of A allele in the STAB2 +1529 C/A locus was significantly associated with SLE (OR=2.2, 95 % CI=1.1~4.2, p=0.016). The genotypic distribution on STAB2 +5207 A/C in SLE and controls did not show significant difference (p=0.057); however, carriage of STAB2 +5207 C allele was positively associated with CNS disorder in SLE patients (OR=5.0, 95 % CI=1.6~15.5, p=0.007). Genotypic variations on STAB2 +1529 and +5207 loci didn’t show significance difference between RA patients and controls. In addition, the haplotype frequency of the STAB2 [phone] C/G-G/T did not show significant difference between SLE or RA patients and their control groups. Moreover, allelic frequency of BAI1 +5010C on BAI1 3’UTR region was positively correlated to both SLE (p=0.007) and RA patients (p=0.019). In the present study, we found several genetic polymorphisms in apoptotic cell clearance related genes were positively associated with autoimmune diseases in Taiwanese population, suggesting apoptotic cell clearance deficiency may be one of the predisposition factor for these diseases. Further study of the altered biological function in STAB2 and BAI1 and the respective genetic polymorphisms would serve to understand and establish them as potential genetic markers for SLE or RA in Taiwanese population.

並列關鍵字

Efferocytosis ； Systemic Lupus Erythematosus (SLE) ； Rheumatoid Arthritis (RA) ； Genetic polymorphisms ； Stabilin-2 ； Bai1

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