介白素-19( Interleukine-19；IL-19)及IL-20與許多發炎反應疾病息息相關，其中IL-19已知與局部發炎反應疾病，例如乾癬以及氣喘相關。而實驗室先前的研究指出，以內毒素(Endotoxin；Lipopolysaccride)誘發小鼠敗血性休克(LPS-induced septic shock)的動物實驗中，IL-19也參與組織傷害的過程；而IL-20已知在乾癬症 (psoriasis)、類風濕性關節炎(Rheumatoid Arthritis)以及動脈粥狀硬化 (Atherosclerosis)等疾病中，參與其中的發炎反應及組織傷害。IL-19以及IL-20皆隸屬於IL-10 family，IL-19與IL-20共用一組接受體複合體傳遞細胞訊息：IL-20R1/IL-20R2，而IL-20還可以透過另一組接受體複合體作用：IL-22R1/ IL-20R2。可溶性受體(soluble receptor)，即細胞外接受體的部份，sIL-20R1以及sIL-20R2 ，仍可與IL-19及IL-20結合，而中和其作用。可表達可溶性受體的質體(soluble IL-20R1 plasmid DNA)以電擊(Electroporation)方式送入小鼠體內，在小鼠敗血性休克以及大鼠類風濕性關節炎的實驗動物中，具有中和IL-19以及IL-20的作用，而達到疾病保護的效果。因此我們想藉由建立soluble receptor基因轉殖鼠，在老鼠可以持續過量表現可溶性受體的情況之下，而不以電擊方式送入老鼠體內產生暫時性可溶性受體蛋白，觀察轉殖鼠對於與IL-19以及IL-20相關發炎疾病中可溶性受體在活體老鼠所扮演的角色。未來在治療此類發炎反應疾病是否能有新的醫藥拓展的可能。將smIL-20R2序列接至pcDNA3.1的載體表現系統，以原核胚顯微注射方式製作基因轉殖鼠，抽取尾巴基因進行聚合酶鏈鎖反應(Polymerase Chain Reaction; PCR )，確定小鼠基因確實嵌入smIL-20R2，基因型(Genotype)確定後，再利用ELISA方式偵測smIL-20R2蛋白質於轉殖鼠血清中的表現，用以確定表現型(Phenotype)。當基因轉殖小鼠確定建立完成後再進一步分析smIL-20R2在in vivo中對老鼠敗血性休克病理機制的影響。在本實驗中，將可溶性受體基因轉殖小鼠(smIL-20R2)應用在內毒素誘發敗血性休克小鼠動物模式之中，在LPS誘發後二十四小時，smIL-20R2基因轉殖小鼠於肺臟、肝臟以及腎臟組織的傷害( 嗜中性白血球浸潤強度 )比起對照組Wild
3
type ( WT )鼠還要來的低；且在小鼠血清經過生化數據的檢測後，發現肝指數GOT及腎功能指標：BUN、Creatinine的數值，在敗血性休克的情況之下，smIL-20R2基因轉殖小鼠比起對照組WT小鼠也相對的低。由此可知，可溶性受體(smIL-20R2)的蛋白，在敗血性休克的過程當中可能藉由中和IL-19，而使得敗血症病程進展過程降低或是減緩LPS對肺臟、肝臟或是腎臟的傷害，結果顯示smIL-20R2對於敗血性休克確實具有保護作用。

IL-19 and IL-20 are associated with many inflammatory diseases. Our previous studies showed that IL-19 was involved in tissue injury in mice model with endotoxin shock. We also found that IL-20 was a pleiotropic cytokine with potent inflammatory, angiogenic, and chemoattractive effects, all of which are characteristics of psoriasis, rheumatoid arthritis, and atherosclerosis. In the IL-10 family, IL-19 and IL-20 shares receptor complexes. IL-19 and IL-20 are capable of signaling through IL-20R1/IL-20R2 heterodimer and IL-20, but not IL-19, can also use IL-22R1/IL-20R2 receptor complex. Soluble form of the extra-cellular domains of IL-20 receptors, sIL-20R1 and sIL-20R2, can be used to neutralize the activities of IL-19 and IL-20 in vitro and in vivo. Therefore, blocking the biological activities of IL-19 and IL-20 through over-expression of smIL-20R1 or smIL-20R2 in transgenic mice will be a new strategy to study the mechanism of IL-19 and IL-20-associated diseases. To select the smIL-20R2 transgenic mice, we analyzed genomic DNA by using PCR with certain primers and performed ELISA to detect the serum level of smIL-20R2 in the mice. After analyzing the genotype and phenotype, smIL20R2 transgenic mice are confirmed to be successfully established. We applied LPS-induced septic shock animal model on sIL-20R2 transgenic mice to investigate whether sIL-20R2 can neutralize IL-19-induced effects and protect vital organs from damage. We found neutrophil infiltration in lung, liver and kidney is decreased in smIL-20R2 transgenic mice compared to control mice. The transcript and protein of sepsis-induced IL-19 in lung tissue was decreased. Twenty-four hours after induction of septic shock, the serum levels of GPT, BUN and Creatinine in smIL-20R2 transgenic mice are lower than those of control mice. In conclusion, over-expression of smIL-20R2 can indeed decreased IL-19 expression and ameliorate neutrophil infiltration in lung, liver and kidney after sepsis induction. Besides, over-expressed smIL-20R2 in transgenic mice can also prevent liver and kidney from damage in the progression of septic shock. Theses results suggest increased smIL-20R2 in transgenic mice can protect lung, liver, kidney from IL-19 mediated tissue damage in septic shock.

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