Adenovirus를 이용한 Uteroglobin 유전자 치료가 실험적 사구체신염에 미치는 영향 (Adenovirus-mediated Gene Therapy of Uteroglobin Protects the Experimental Glomerulonephritis in Mice)

서지정보

· 발행기관 : 대한신장학회
· 수록지 정보 : Kidney Research and Clinical Practice / 24권 / 2호 / 181 ~ 190페이지
· 저자명 : 김현리, 양승희, 성은영, 이정은, 이동섭, 정종훈, 김연수

초록

목 적:저자들은 면역 조절 및 항염증작용을 가진 Uteroglobin (UG)이 면역복합체의 신장 내 침착을 억제하여 사구체신염의 발생을 억제함을 보고한 바 있다. 본 연구에서는 항기저막 사구체신염 (anti-GBM GN) 생쥐모델에서 아데노바이러스를 이용한 UG 유전자의 신장 내 발현이 사구체신염의 발생에 미치는 영향을 평가하고자 하였다.
방 법:아데노바이러스 벡터를 이용한 UG 발현 아데노바이러스 (AdCMV-UG)를 제조하여 8주령의 생쥐 (C57BL/6)의 신장에 주입 후 항기저막 사구체신염 (anti-GBM GN)을 유도하여 신염의 정도를 비교하였다. UG 치료군 (AdCMV-UG)과 질환 대조군 (AdCMV-GFP, PBS only)을 대상으로 anti-GBM GN 유도 후 7일과 14일로 나누어 단백뇨와 신장의 형태학적 변화를 관찰하였다.
결 과:UG 유전자와 단백질은 신장조직과 메산지움 세포에서 발현하였다. 신염 유도 7일째 (총 12일) AdCMV-UG 주입군은 질환 대조군 (AdCMV-GFP, PBS only)에 비해 요단백/ creatinine 값이 유의하게 감소하였으며 (p=0.001), 조직학적으로도 사구체 메산지움 증식과 세포성 반월상 역시 대조군에 비해 감소된 결과를 보였다. 그러나, 신염 유도 14일 (총 19일)로 갈수록 단백뇨나 형태학적으로 UG의 효과는 감소되었다.

영어초록

Purpose:Uteroglobin (UG), steroid inducible cytokine-like protein, has potent anti-inflammatory and immunomodulatory action. It is secreted by the mucosal epithelia of virtually all mammals. The aims of this study were to investigate the efficacy of recombinant adenovirus carrying uteroglobin (AdCMV-UG) in prevention and treatment of glomerulonephritis (GN) in mice.
Methods:The AdCMV-UG was created by inserting the uteroglobin cDNA into the pAdTrack- CMV vector and was transfected into the 293 cells through liposome mediated vehicles. AdCMV-UG was injected direct to the both kidneys of 20 mice. In control groups (disease controls), 13 mice received adenoviral vector with GFP and another 11 mice received PBS only. After 5days of viral injection, GN was induced by repetitive intravenous injection of 3.0 mg rabbit anti-GBM Ab to the pretreated mice (C57/B6). Histological and biochemical changes were evaluated 7 and 14 days after injection of anti-GBM Ab.
Results:UG was expressed in the renal tissues and mesangial cells infected with the infection of AdCMV-UG. Pretreatment with AdCMV-UG attenuated the cellular crescent formation 7 days after induction of GN when compared to AdCMV-GFP, PBS only. We also observed reduced mesangial matrix expansion in mice treated with adenovirus carrying UG. Proteinuria was significantly reduced in the mice treated with adenovirus carrying UG when compared with disease control mice (AdCMV-UG 102.2±20.97, AdCMV-GFP 170.6±41.77, and PBS 169.8±55.67, respectively p<0.05 mg/mg). However, at 14 days after anti-GBM Ab injection (total 19 days), there was no significant difference in the amounts of proteinuria and morphologic findings between pretreated and disease control groups.
Conclusion:Adenoviral mediated gene transfer is an effective way of gene delivery. Locally expressed uteroglobin attenuated the severity of glomerulonephritis induced by anti-GBM antibody, although it was transient. Gene therapy using uteroglobin may be constituted for the treatment of human diseases such as chronic GN.

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