Adenovirus-mediated antisense ERK2 gene therapy ameliorates chronic allograft nephropathy in a rat model

doi:10.1007/s11684-009-0039-0

Front Med Chin

2009, Vol. 3

Issue (2) : 204-210 https://doi.org/10.1007/s11684-009-0039-0

RESEARCH ARTICLE

Adenovirus-mediated antisense ERK2 gene therapy ameliorates chronic allograft nephropathy in a rat model

Zhao DING, Zhishui CHEN, Xilin CHEN, Ming CAI, Hui GUO, Nianqiao GONG(

)

Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

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Abstract

To investigate the effect and underlying mechanism of adenovirus-mediated antisense ERK2 (Adanti-ERK2) gene therapy upon chronic allograft nephropathy (CAN) of rats, male Lewis (LEW, RT11) rats received male Fisher (F344, RT11v1) renal allografts. The recipients were divided into three groups: (1) empty control group; (2) vector control group; (3) gene therapy group. All recipients were sacrificed for the grafts and serum analysis at the 24th week after transplantation. Morphometric analysis was used to determine the fibrosis of grafts. Immunohistochemistry was used to detect the expression of E-Cadherin, Vimentin, TβR I and the infiltration of CD4⁺ T lymphocyte, CD8⁺ T lymphocyte and ED-1⁺ monocytes. Enzyme linked immunosorbent assay (ELISA) was used to detect TGF-β1 in serum. The grafts in the control group and vector control group showed CAN. There was less E-Cadherin in renal tubular epithelial cells in the empty control group but more Vimentin and TβR I. In the gene therapy group, the fibrosis was ameliorated and fewer T lymphocytes and ED-1⁺ monocytes infiltrated in the interstitium. There was no significant difference in the expression of E-Cadherin between the gene therapy group and normal rats. Compared with the empty control group, the expression of TGF-β1 in the gene therapy group was down-regulated. Adanti-ERK2 gene therapy protects the renal allograft and attenuates graft fibrosis, which may be correlated with a decreased renal tubular epithelial mesenchymal transition, a decreased infiltration of CD4⁺ T lymphocyte, CD8⁺ T lymphocytes and ED-1⁺ monocytes in renal interstitium, and the down-regulated TGF-β1 expression.

Keywords anti-ERK2 renal transplantation epithelial mesenchymal transition chronic allograft nephropathy

Corresponding Author(s): GONG Nianqiao,Email:nqgong@tjh.tjmu.edu.cn

Issue Date: 05 June 2009

Cite this article:

Zhao DING,Zhishui CHEN,Xilin CHEN, et al. Adenovirus-mediated antisense ERK2 gene therapy ameliorates chronic allograft nephropathy in a rat model[J]. Front Med Chin, 2009, 3(2): 204-210.

URL:

https://academic.hep.com.cn/fmd/EN/10.1007/s11684-009-0039-0
https://academic.hep.com.cn/fmd/EN/Y2009/V3/I2/204

Fig.1 HK-2 cells were transfected with Adanti-ERK2 for 72 h. The whole cell lysate was immunoblotted with specific ERK2 antibody. The ERK2 protein was down-regulated by Adanti-ERK2 gene therapy.

Fig.2 The grafts were harvested for histology and morphometry at the 24th week after transplantation. Tubular atrophy, vascular occlusive changes, glomerulosclerosis (GS) and chronic interstitial fibrosis appeared in the grafts of the empty control group and vector control group. The morphologies of the grafts in the gene therapy group were normal (HE staining, × 400).

Fig.3 Immunohistochemical staining showed the infiltrating inflammatory cells in the allografts 24 weeks after transplantation. In the gene therapy group, the infiltrations of ED-1 macrophages, CD8 T and CD4 T lymphocytes were less than those in the empty control and the vector contol group (× 400).

Tab.1 Adanti-ERK2 modulated infiltrations of CD8 T, CD4 T lymphocytes and macrophages

Fig.4 gene transfer of Adanti-ERK2 to rat renal allograft attenuated tubular epithelial-myofibroblast transition. At the 24th week after transplantation, Paraformaldehyde-fixed paraffin sections of the grafts were stained with immunohistochemistry to measure the E-Cadherin and Vimentin with mouse anti-rat E-Cadherin and Vimentin antibodies. The renal tubular epithelial cells of grafts in the control group and vector control group expressed less E-Cadherin but more Vimentin and α-SMA in comparison with the gene therapy group (× 400).

Fig.5 Adanti-ERK2 suppressed TβR I up-regulation in chronic allograft nephropathy. At the 24th week after transplantation, paraformaldehyde-fixed and paraffin-embedded sections of the grafts were stained with immunohistochemistry to detect TβR I with mouse anti-rat TβR I antibody (1∶100). In the empty control group and vector control group, the renal tubular epithelial cells expressing TβR I were more than those in the gene therapy group, indicating the Adanti-ERK2 inhibited the up-regulation of TβR I (× 400).

Tab.2 Adanti-ERK2 down-regulated the expression of TGF-β1 and suppressed TβR I up-regulation in chronic allograft nephropathy

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