源自HIV-1转基因小鼠的系膜细胞增殖、凋亡及基质积累增强。

Enhanced proliferation, apoptosis, and matrix accumulation by mesangial cells derived from HIV-1 transgenic mice.

作者信息

Singhal P C, Sharma P, Loona R, Gibbons N, Franki N, Klotman P E

机构信息

Department of Medicine, Long Island Jewish Medical Center, New Hyde Park, NY 11040, USA.

出版信息

J Investig Med. 1998 Aug;46(6):297-302.

PMID:9737092

Abstract

BACKGROUND

Mice, transgenic for HIV-1 genes, have been demonstrated to develop renal lesions mimicking HIV-associated nephropathy. Focal glomerulosclerosis (FGS) has been reported to be the predominant glomerular lesion in these animals. In the other models of FGS, the accumulation of mesangial matrix and mesangial cell proliferation have been shown to be the preceding abnormalities. We evaluated the proliferation, apoptosis, and matrix accumulation by mesangial cells derived from mice transgenic for HIV-1 genes as well as from nontransgenic mice.

METHODS

Mesangial cells were cultured from mice transgenic for HIV-1 genes (HTrMC) and nontransgenic mice (NTrMC) of the same age and sex. The growth rate of HTrMC and NTrMC was determined under identical conditions. Morphologic evaluation of apoptosis was performed by staining cells with Hoechst (H)-33342 and propidium iodide. Accumulation of mesangial cell collagen type IV, laminin, and fibronectin was measured by the dot blot assay. Total RNA was extracted from HTrMC and NTrMC and Northern blots were generated. These blots were probed with specific probes for TGF-beta, proteoglycan (P16), and GAPDH.

RESULTS

Mesangial cells (HTrMC) derived from transgenic mice had greater (P < 0.004) proliferation when compared to mesangial cells (NTrMCs) from nontransgenic mice (HTrMCs, 4.2 +/- 0.3 vs NTrMCs, 3.0 +/- 0.2 x 10(4) cells/well). HTrMCs also showed enhanced (P < 0.0001) apoptosis compared to NTrMCs (HTrMCs, 13.2 +/- 1.5% vs NTrMCs, 3.1 +/- 0.5% apoptotic cells/field). HTrMCs accumulated an increased (P < 0.02) amount of collagen type IV (HTrMCs, 5659.7 +/- 472.8 vs NTrMCs, 3882.2 +/- 339.7 ng/well); whereas NTrMCs accumulated a greater amount of laminin when compared to HTrMCs (HTrMCs, 12.8 vs NTrMCs, 29.6 +/- 2.9 ng/well). HTrMCs also showed an enhanced mRNA expression of TGF-beta and an attenuated expression of proteoglycan (P16).

CONCLUSIONS

These results suggest that mesangial cells derived from mice transgenic for HIV-1 genes have enhanced proliferation and collagen accumulation. The enhanced expression of TGF-beta may have contributed to enhanced HTrMC proliferation and the accumulation of collagen. The present study provides the basis for a hypothesis that mesangial cells may be contributing to the development of focal glomerulosclerosis in mice transgenic for HIV-1 genes.

摘要

背景

已证实携带HIV-1基因的转基因小鼠会出现类似HIV相关性肾病的肾脏病变。据报道，局灶性肾小球硬化（FGS）是这些动物主要的肾小球病变。在其他FGS模型中，系膜基质积聚和系膜细胞增殖已被证明是先前存在的异常情况。我们评估了来自携带HIV-1基因的转基因小鼠以及非转基因小鼠的系膜细胞的增殖、凋亡和基质积聚情况。

方法

从相同年龄和性别的携带HIV-1基因的转基因小鼠（HTrMC）和非转基因小鼠（NTrMC）中培养系膜细胞。在相同条件下测定HTrMC和NTrMC的生长速率。通过用Hoechst（H）-33342和碘化丙啶对细胞进行染色来进行凋亡的形态学评估。通过斑点印迹法测定系膜细胞IV型胶原、层粘连蛋白和纤连蛋白的积聚情况。从HTrMC和NTrMC中提取总RNA并进行Northern印迹分析。用针对转化生长因子-β（TGF-β）、蛋白聚糖（P16）和甘油醛-3-磷酸脱氢酶（GAPDH）的特异性探针检测这些印迹。

结果

与来自非转基因小鼠的系膜细胞（NTrMC）相比，来自转基因小鼠的系膜细胞（HTrMC）具有更高的增殖率（P < 0.004）（HTrMC为4.2±0.3×10⁴个细胞/孔，NTrMC为3.0±0.2×10⁴个细胞/孔）。与NTrMC相比，HTrMC也表现出增强的凋亡（P < 0.0001）（HTrMC为13.2±1.5%凋亡细胞/视野，NTrMC为3.1±0.5%凋亡细胞/视野）。HTrMC积聚了更多（P < 0.02）的IV型胶原（HTrMC为5659.7±472.8 ng/孔，NTrMC为3882.2±339.7 ng/孔）；而与HTrMC相比，NTrMC积聚了更多的层粘连蛋白（HTrMC为12.8，NTrMC为29.6±2.9 ng/孔）。HTrMC还表现出TGF-β mRNA表达增强和蛋白聚糖（P16）表达减弱。