Hepatocyte growth factor gene therapy retards the progression of chronic obstructive nephropathy.
Hepatocyte growth factor gene therapy retards the progression of chronic obstructive nephropathy.
复制标题
肝细胞生长因子基因治疗可延缓慢性阻塞性肾病的进展。
作者：
Xiaojie Gao;H. Mae;N. Ayabe;T. Takai;Keisuke Oshima;M. Hattori;T. Ueki;J. Fujimoto;T. Tanizawa
背景
单侧输尿管梗阻(UUO)以进行性肾小管萎缩和间质纤维化为特征。细胞增殖和凋亡之间平衡的打破在肾萎缩中起着关键作用。肝细胞生长因子(HGF)是一种对细胞存活和组织再生起作用的细胞因子。我们研究了肝细胞生长因子基因治疗对慢性梗阻性肾病肾小管细胞存活和抗纤维化的影响及其可能机制。
方法
用含日本血凝病毒脂质体(HVJ Liposome)对UUO大鼠进行了体内反复转导HGF基因骨骼肌的实验。用原位杂交、酶联免疫吸附试验或免疫组织化学方法检测肝细胞生长因子和c-Met的表达。用Masson‘s三色染色、α-平滑肌肌动蛋白和ED-1免疫组织化学染色检测肾间质纤维化和巨噬细胞浸润情况。免疫组织化学法和Western blotts法检测细胞增殖细胞核抗原(PCNA)、Bcl2、Bclxl和Bax等细胞存活指数。TUNEL法检测细胞凋亡率。
结果
转导HVJ-HGF基因后，内源性HGF和c-Met在UUO肾脏中表达上调。肾纤维化、巨噬细胞浸润和肾小管萎缩在UUO后第14天和第28天均受到抑制(P<；0.05或0.01)。UUO患者肾小管上皮细胞增殖活跃，细胞凋亡受到抑制，尤以UUO晚期为甚。转染HGF的UUO大鼠BCL-2表达增强，而Bclxl和Bax表达无明显变化。
结论
体内HGF基因转染可延缓慢性梗阻性肾病的进展，保护UUO模型中肾小管细胞的存活。BCL-2而不是BclxL或Bax可能参与了HGF的抗凋亡作用。
BACKGROUND
Unilateral ureteral obstruction (UUO) is characterized by progressive tubular atrophy and interstitial fibrosis. Rupture of the balance between cell proliferation and apoptosis plays a critical role in renal atrophy. Hepatocyte growth factor (HGF) is a cytokine function on cell survival and tissue regeneration. We studied the effects and possible mechanisms of HGF gene therapy on tubular cell survival and anti-fibrosis in chronic obstructed nephropathy.
METHODS
An in vivo transfection procedure of repeatedly transducing skeletal muscles with the HGF gene using liposomes containing the hemagglutinating virus of Japan (HVJ liposome) was tested on UUO rats. Expression of HGF and c-Met were examined by in situ hybridization, ELISA, or immunohistochemical staining. Interstitial fibrosis and macrophage infiltration were evaluated by Masson's Trichrome staining, alpha-smooth muscle actin and ED-1 immunostaining. Cell survival indices including proliferating cell nuclear antigen (PCNA), Bcl-2, Bcl-xL and Bax were measured by immunohistochemistry and Western blots. Apoptosis was determined by the TUNEL method.
RESULTS
After HVJ-HGF gene transfer, endogenous HGF and c-Met were up-regulated in UUO kidneys. Renal fibrosis, macrophage infiltration and tubular atrophy were suppressed both at day 14 and 28 after UUO (P < 0.05 or 0.01). Tubular cell proliferation was activated while apoptosis was inhibited, especially at the late stage of UUO. Bcl-2 was enhanced in the HGF-transfected UUO rats, while no changes of Bcl-xL and Bax were found.
CONCLUSIONS
In vivo HGF gene transfection retards the progression of chronic obstructed nephropathy and protects tubular cell survival in the long-term UUO model. Bcl-2 rather than Bcl-xL or Bax may contribute to the anti-apoptotic function of HGF.
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肝细胞生长因子受体 c-met 的信号转导。
DOI：
10.1681/asn.v5111872
发表时间：
1995
期刊：
Journal of the American Society of Nephrology : JASN
影响因子：
--
作者：
Cantley,LG;Cantley,LC
通讯作者：
Cantley,LC
影响因子：
19.6
作者：
Chevalier, RL;Kim, A;Wolstenholme, JT
通讯作者：
Wolstenholme, JT
DOI：
10.1152/ajprenal.1999.277.4.f624
发表时间：
1999-10-01
影响因子：
4.2
作者：
Liu, YH
通讯作者：
Liu, YH
影响因子：
--
作者：
MILLER, SB;MARTIN, DR;HAMMERMAN, MR
通讯作者：
HAMMERMAN, MR
影响因子：
19.6
作者：
Liu, YH;Rajur, K;Dworkin, LD
通讯作者：
Dworkin, LD