[核心蛋白聚糖转染系膜细胞载体注射对大鼠抗 Thy-1 血清诱导肾炎的拮抗作用]

[The antagonistic effect on anti-thy-1 serum-induced nephritis of rats injected by decorin-transfected mesangial cells vector].

作者信息

Wang Hui-jun, Zhang Zhi-gang, Liu Xue-guang, Zhang Xiu-rong, Chen Qi, Guo Mu-yi

机构信息

Department of Pathology, Shanghai Medical College, Fudan University, Shanghai 200032, China.

出版信息

Zhonghua Bing Li Xue Za Zhi. 2003 Oct;32(5):444-8.

PMID:14633458

Abstract

OBJECTIVES

To inject decorin-transfected mesangial cells (MsC) vector into the kidneys of rats with anti-thy-1 serum-induced nephritis via left renal artery and observe the survival condition of MsC vector and its influence on glomerular lesions in rats with anti-thy-1 serum induced nephritis.

METHODS

Rat mesangio-proliferative glomerulonephritis was established by tail intravenous injection with rabbit anti-thy-1 serum (ATS). Decorin-transfected MsC was injected into rat kidneys via left renal artery. Primary culture, immunostaining for BrdU and decorin of transfected MsC lines were performed to observe their survival. Immunohistochemistry with image analysis was performed to detect the expression of BrdU, alpha-SMA, decorin, TGF-beta1, FN and ColIV in diseased glomeruli.

RESULTS

Rat anti-thy-1 serum-induced nephritis identified by pathological examination was successfully established by injecting rabbit ATS, and decorin transfected MsC vector was transfused to rat glomeruli via left renal artery. The active growth and positive expressions of BrdU and decorin proteins on the nuclei and cytoplasms of ex vivo MsC were observed respectively. TGF-beta1, FN, ColIV expressions in diseased glomeruli of rats with ATS nephritis were decreased significantly at day 4 (TGF-beta1, P < 0.05) and day 2 (FN and ColIV, P < 0.01) respectively, compared to uninjected kidneys.

CONCLUSIONS

MsC vector is successfully transferred to the glomeruli of experimental rats via left renal artery injection with no affect on cell survival. Decorin protein is expressed on the transfected MsC and shows antagonistic effect on the glomerular lesions of ATS rats. It suggests that the use of ex vivo MsC vector system can provide useful experimental basis for gene therapy of kidney disease in animal model.

摘要

目的

通过左肾动脉将转染了核心蛋白聚糖的系膜细胞（MsC）载体注入抗 Thy-1 血清诱导的肾炎大鼠肾脏，观察 MsC 载体的存活情况及其对抗 Thy-1 血清诱导的肾炎大鼠肾小球病变的影响。

方法

通过尾静脉注射兔抗 Thy-1 血清（ATS）建立大鼠系膜增生性肾小球肾炎模型。将转染了核心蛋白聚糖的 MsC 经左肾动脉注入大鼠肾脏。对转染的 MsC 系进行原代培养、BrdU 和核心蛋白聚糖免疫染色以观察其存活情况。采用免疫组织化学及图像分析检测病变肾小球中 BrdU、α-SMA、核心蛋白聚糖、转化生长因子-β1（TGF-β1）、纤维连接蛋白（FN）和Ⅳ型胶原（ColIV）的表达。

结果

经病理检查证实，注射兔 ATS 成功建立了大鼠抗 Thy-1 血清诱导的肾炎模型，转染了核心蛋白聚糖的 MsC 载体经左肾动脉输注至大鼠肾小球。分别观察到体外培养的 MsC 细胞核和细胞质上 BrdU 及核心蛋白聚糖蛋白的活跃生长和阳性表达。与未注射的肾脏相比，ATS 肾炎大鼠病变肾小球中 TGF-β1、FN、ColIV 的表达在第 4 天（TGF-β1，P<0.05）和第 2 天（FN 和 ColIV，P<0.01）分别显著降低。