Hepatocyte growth factor gene therapy retards the progression of chronic obstructive nephropathy

肝细胞生长因子基因治疗延缓慢性阻塞性肾病的进展

• 批准号: 12670794
• 负责人: TANIZAWA Takakuni
• 金额: $ 2.18万
• 依托单位: Hyogo College of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670794/
• 关键词: HGF; Renal fibrosis; Gene therapy; UUO; chronic obstructive nephropathy; 腎線維化; 遺伝子治療; 一側尿管結紮ラット; 肝細胞成長因子; 腎間質線維化; 一側尿管結紮モデル

Glomerular sclerosis and tubulo-interstitial fibrosis are the common cause of end-stage renal failure and are supposed to be irreversible, regardless of the primary etiology of renal disease. Hepatocyte growth factor (HGF) is now well known as a cytokine with numerous functions on diverse cell survival by inhibiting apoptosis and tissue regeneration. Compared with repeatedly injection of HGF protein, gene therapy is the best for its efficiency and financial economy. In this study, we studied the effects and possible mechanisms of HGF gene therapy on tubular cell survival and blockage of renal fibrosis in chronic obstructed nephropathy by unilateral ureteral obstruction (UUO).An in vivo transfection procedure of repeatedly transducing skeletal muscles with HGF gene using liposomes containing the hemagglutinating virus of Japan (HVJ liposome) was tested on UUO rats. Thirty 5-week-old male Sprague-Dawley rats were subjected to complete UUO or Sham operation. UUO rats were randomly separat … More ed into two groups. The treated group received repetitive intramuscular injection of HVJ liposomes containing 800μg pUC-SRα/HGF once a week, while the untreated group was injected with BSS. Then kidney tissues were collected for further assay at 2w or 4w after operations. Expression of HGF and c-Met were examined by in situ hybridization. Interstitial fibrosis and macrophage infiltration were evaluated by Masson's Trichrome staining, α-SMA and ED-1 immunostaining. Cell survival indices including PCNA, Bcl-2, Bcl-xL and Bax were measured by immunohistochemistry and western blots. Apoptosis was determined by TUNEL method.After HGF-HVJ-liposome gene transfer, endogenous HGF and c-Met were up regulated. Renal interstitial volume and fibroblast activity were suppressed both at day 14 and 28 after UUO (p<0.05 or 0.01). The tubular macrophage infiltration in the late-stage obstructed kidneys was significantly reduced. Tubular atrophy progression after the obstruction was also suppressed. Tubular cell proliferation was activated while apoptosis was inhibited, especially at the late-stage of UUO. Bcl-2 was enhanced in HGF transfected UUO rats, while no changes of Bcl-xL and Bax were found. Less

肾小球硬化症和tubo Interstitial纤维化是终末期肾衰竭的常见原因，而不论肾脏疾病的主要病因学如何，预计将是不可逆的。肝细胞生长因子（HGF）现在众所周知是一种细胞因子，通过抑制细胞凋亡和组织再生，在潜水细胞存活方面具有众多功能。与反复注射HGF蛋白相比，基因疗法是其效率和金融经济的最佳选择。在这项研究中，我们研究了HGF基因治疗对慢性骨质障碍物（UUO）的慢性阻塞肾病（UUO）的肾脏纤维化的肾脏纤维化的影响和可能的机制。一种反复传递HGF基因的体内转染程序。在UUO大鼠上测试了脂质体）。三十五周大的雄性Sprague-Dawley大鼠接受了完整的UUO或假手术。 UUO大鼠被随机分离……更多地分为两组。治疗组每周接受一次重复的肌内注射HVJ脂质体，每周一次含有800μgPUC-SRα/HGF，而未处理的组则注入了BSS。然后，在手术后，收集肾脏组织在2W或4W下进行进一步测定。通过原位杂交检查HGF和C-MET的表达。通过Masson的三色染色，α-SMA和ED-1免疫染色评估间质纤维化和巨噬细胞浸润。通过免疫组织化学和蛋白质印迹测量了包括PCNA，BCL-2，BCL-XL和BAX在内的细胞存活指数。通过TUNEL方法确定凋亡。在HGF-HVJ-脂体基因转移后，内源性HGF和C-MET受到调节。 UUO后第14和28天都抑制肾脏间质量和成纤维细胞活性（P <0.05或0.01）。晚期肾脏阻塞的肾小管巨噬细胞浸润显着降低。梗阻后的管状萎缩进展也被抑制。在抑制凋亡的同时激活了管状细胞增殖，尤其是在UUO晚期。 HGF翻译的UUO大鼠增强了Bcl-2，而没有发现BCL-XL和BAX的变化。较少的

项目成果

Tanizawa , Takai T: "Medical management for pediatric nephrotic syndrome"Shonikarinnsyo. 54. 615-620 (2001)

谷泽，高井 T：“小儿肾病综合征的医疗管理”Shonikarinnsyo。

Minagawa K, Tsuji Y, Ueda H, Koyama K, Tanizawa T, Okamura H, Tamaoki-Hashimoto T: "Possible correlation between high levels of IL-18 in the cord blood of pre-term infants and neonatal development of periventricular leukomalacia and cerebral palsy"CYTOKIN

Minakawa K、Tsuji Y、Ueda H、Koyama K、Tanizawa T、Okamura H、Tamaoki-Hashimoto T：“早产儿脐带血中高水平的 IL-18 与新生儿脑室周围白质软化和大脑发育之间可能存在相关性”

Mae H, Gao X, Takai T, Ooshima K, Hattori M, Tanizawa T: "A case of children with poststreptococcal acuteglomerulonephritis associated with acute renal failure"J Jap Soc Pediatr Renal failure. 22. 62-65 (2002)

Mae H、Gao X、Takai T、Ooshima K、Hattori M、Tanizawa T：“链球菌后急性肾小球肾炎伴急性肾衰竭的儿童病例”J Jap Soc Pediatr Renal failure。

Asano Y, Kaneda Y, Takeda H, Konishi M, Ootsuka M, Ootsuki F, Tsujino Y, Yamsmoto M, Tmizawa T: "QOL in children with aplastic anemia after therapy. Comparison between bone marrow transplantation and multiple chemotherapy"Jap J Pediatr Hematol. 16. 62-70

Asano Y、Kaneda Y、Takeda H、Konishi M、Ootsuka M、Ootsuki F、Tsujino Y、Yamsmoto M、Tmizawa T：“再生障碍性贫血儿童治疗后的生活质量。骨髓移植与多种化疗的比较”Jap J Pediatr Hematol

Kaneda Y, Okamoto N, Osako T, Maeda H, Tokuyama M, Takenaka Y, Tanizawa T: "A case of intractable nephrotic syndrome with successful treatment by LDL apheresis"Jap J Pediatr Nephrol. 15. 49-54 (2002)

Kaneda Y、Okamoto N、Osako T、Maeda H、Tokuyama M、Takenaka Y、Tanizawa T：“通过 LDL 单采术成功治疗顽固性肾病综合征的病例”Jap J Pediatr Nephrol。