臺灣博碩士論文加值系統

本論文研究之主要目的在於驗證內源性（Endogenous）之半乳糖凝集
素-3(Galectin-3; Gal-3)在狼瘡腎炎(Lupus nephritis)中所扮演
腎臟的保護角色及相關之分子機制。我們近期相關研究之成果顯示，
NLRP3 發炎體與自嗜作用在狼瘡腎炎之發生與惡化皆扮演一關鍵性角
色。本論文之研究結果顯示：[1]狼瘡腎炎病人之腎臟切片具有Gal-
3 蛋白質之表現；[2] Gal-3 基因剔除小鼠(Gal-3 knockout mice)誘
發類狼瘡腎炎模式具嚴重之腎臟病理變化 ；[3]經由Gal-3 基因剔除
小鼠與自發型狼瘡腎炎KO1.Yaa 小鼠之配種，成功建立一全新自發型
之惡化狼瘡腎炎小鼠（Gal-3 KO-KO1.Yaa mice）模式，較背景鼠具
嚴重腎臟病 理與免疫之變化。據此，推測內源性Gal-3 對狼瘡腎炎
具腎臟保護之功能，值得進 一步探究相關之作用機制。因此進一步
驗證內源性Gal-3 可經由抑制NLRP3 發炎體活化及調控自嗜作用之
裂解NLRP3 發炎體機制，進而抑制巨噬細胞活化與調節T 細胞功能，
抑阻狼瘡腎炎之發生與惡化。研究結果顯示:[1]內源性Gal-3 對狼瘡
腎炎具保護角色；[2]內源性Gal-3 調控對狼瘡腎炎之NLRP3 發炎體
活化及自嗜作。本論文研究驗證與確認了內源性Gal-3 與相關路徑
所調節/ 緩解狼瘡腎炎發生與疾病惡化之分子機制。

Lupus nephritis (LN) is a major complication of systemic lupus
erythematosus. Current therapeutic regimens to control the disease
progression of the acute onset of severe LN include high doses of
corticosteroids, cytotoxic agents and disease-modifying antirheumatic
drugs. However, the therapeutic effects are still insufficient, and there is
still a major concern about the potential adverse events of these drugs. Thus,
the development of new therapeutic regimens with much fewer and more
tolerable side effects is clinically warranted. Galectin-3 (Gal-3) is a β-
galactoside-binding protein and implicated in diverse biological processes
in macrophages, dendritic cells, activated lymphocytes, and epithelial cells.
However, the role of Gal-3 and exact mechanisms involved in the
development and progression/deterioration of LN has yet to be determined,
although it has been shown to express in the renal of patients with LN. The
results show that [1] the distribution and levels of Gal-3 protein were
increased in renal biopsy specimen of patients with LN, compared to
normal control subjects, although there was not significant difference in
Gal-3 protein level in the glomerulus, tubular epithelial cells or periglomerular
infiltrating leukocytes among the several classes of the renal
condition – IIIc, IVa, IVb, IV c and V; [2] successful crossing of Gal-3
knockout (KO) mice with KO1.Yaa mice, a spontaneous mouse model of
LN, to generate Gal-3 KO-KO1.Yaa mice, a new mouse model of
accelerated and severe LN; and [3] deficiency of endogenous Gal-3
resulted in markedly increased severity in clinical and pathological
alterations in both the LPS-induced LN in Gal-3 KO mice and spontaneous
VIII
LN in Gal-3 KOKO1.Yaa mice, compared to their respective wild type
mice that equally induced or spontaneously developed the renal condition.
With the results to be obtained, we are committed to develop a novel
therapeutic approach using Gal-3 plasmids via an ultrasound-mediated
microbubble gene transfer system as our future work, and such a novel
nucleic acid drug candidate may be considered for the therapeutic
intervention for LN in the foreseeable near future.

正文目錄 ..................................................................................................... I
表目錄 ...................................................................................................... IV
圖目錄 ........................................................................................................ V
中文摘要 .................................................................................................. VI
英文摘要 ................................................................................................. VII
第一章 緒言............................................................................................... 1
第一節 紅斑性狼瘡（Systemic Lupus Erythematosus, SLE） ..... 1
第二節 狼瘡腎炎( Lupus nephritis, LN) .......................................... 2
第三節 半乳醣凝集素-3（Galectin-3 , Gal-3） .............................. 7
第四節 狼瘡腎炎中的半乳醣凝集素-3 ............................................. 8
第五節 研究目的 ................................................................................ 9
第二章 材料與方法 ................................................................................ 10
第一節 腎臟病理組織型態學 .......................................................... 10
第二節 用腎臟組織蠟塊染C3, IgG 螢光染色 ............................... 11
第三節 特殊染色Periodic Acid Methenamine Silver (PAM) ...... 12
第四節 免疫組織化學染色 .............................................................. 14
第五節 誘發狼瘡腎炎動物模式 ...................................................... 15

第六節 血清中自體抗體及補體之偵測 .......................................... 15
第七節 統計分析 .............................................................................. 16
第三章 結果............................................................................................. 17
第一節 狼瘡腎炎病人之腎臟切片具有半乳糖凝集素-3 蛋白質之
表現 ...................................................................................... 17
第二節 半乳糖凝集素-3 基因剔除小鼠誘發類狼瘡腎炎模式具嚴
重之腎臟病理變化 .............................................................. 17
第三節 自發型半乳糖凝集素-3 基因剔除之自發型狼瘡腎炎小鼠
模式建立 .............................................................................. 19
第四章 討論............................................................................................. 21
第一節 全身免疫調節 ...................................................................... 21
第二節 於半乳糖凝集素-3 與類狼瘡腎炎之發炎反應的相關性 . 21
第三節 半乳糖凝集素-3 於自發型狼瘡腎炎是否具保護作用 ...... 22
第五章 結論............................................................................................. 24
第六章、參考文獻 .................................................................................. 25
表 ............................................................................................................... 33
圖 ............................................................................................................... 35

表目錄
頁次
表一、世界衛生組織World Health Organization (WHO)狼瘡腎炎的形
態學分類(西元1982 修訂) .......................................................... 33
表二、LN 的急性與慢性評估(NIH: National Institutes of Health) ...... 34

圖目錄
頁次
圖一、偵測半乳醣凝集素-3 在狼瘡腎炎病患之腎臟組織病理學與免疫
組織化學染色表現 .................................................................................. 35
圖二、利用免疫組織化學染色偵測半乳醣凝集素-3 在各類別狼瘡腎炎
患者腎臟組織之表現 .............................................................................. 36
圖三、偵側血液中半乳醣凝集素-3 和補體3、4 和dsDNA 相關性分析
................................................................................................................... 37
圖四、類狼瘡腎炎小鼠之蛋白尿和腎功能的評估 .............................. 38
圖五、類狼瘡腎炎小鼠之腎臟組織病理學分析 .................................. 39
圖六、以免疫組織化學偵測類狼瘡腎炎小鼠之腎臟組織pan-T 細胞浸
潤 ............................................................................................................... 40
圖七、以免疫組織化學染色偵測類狼瘡腎炎小鼠中的腎臟巨噬細浸潤
胞 ............................................................................................................... 41
圖八、以免疫螢光染色確認自發型狼瘡腎炎小鼠之腎組織中免疫複合
物IgG 及C3 之沉積 ................................................................................ 42
圖九、培育半乳醣凝集素-3 基因剔除之自發型狼瘡腎炎小鼠模式 .. 43
圖十、偵測自發型狼瘡腎炎小鼠中蛋白尿及腎功能的曲線圖 .......... 44
圖十一、偵測自發型狼瘡腎炎小鼠之腎臟組織病理染色 .................. 45

圖十二、偵測自發行狼瘡腎炎小鼠之腎臟組織病理學半定量分析 .. 46
圖十三、以Periodic Acid Methenarnine(PAM)染色分析自發型狼瘡腎
炎小鼠腎臟組織纖維化之情形 .............................................................. 47
圖十四、分析自發型狼瘡腎炎小鼠腎臟組織之CD3 T 細胞浸潤情形
................................................................................................................... 48
圖十五、CD3 T 細胞之半定量分析 ..................................................... 49
圖十六、以免疫化學染色偵測自發型狼瘡腎炎小鼠腎臟組織之巨噬細
胞（F4/80）浸潤情形 ............................................................................. 50
圖十七、巨噬細胞之半定量分析 ........................................................ 51

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