Alpha-Actinin-4 is a Possible Target Protein for Aristolochic Acid I in Human Kidney Cells In Vitro

Aristolochic acid I (AA-I) is a strong nephrotoxin, carcinogen, and mutagen found in plants such as the Aristolochia species. The mechanisms underlying AA-I toxicity in the kidneys are poorly understood. In this study, we aimed to gain insight into the mechanism of AA-I nephrotoxicity by analyzing the uptake, subcellular distribution, and intracellular targets of AA-I in the human kidney cell line HK-2 using immunocytochemistry, immunoprecipitation, and LC-MS/MS. In HK-2 cells incubated with 20[Formula: see text][Formula: see text]g/mL AA-I for different periods of time (up to 12[Formula: see text]h), AA-I was detected by a specific monoclonal antibody (MAb) against AA-I, both in the cytoplasm and nuclei. Nuclear localization depended on the exposure time. A protein with the molecular weight of 100 kDa was immunoprecipitated with the anti-AA-I MAb from the AA-I-treated cell lysates and was identified by LC-MS/MS as [Formula: see text]-actinin-4 after digestion of the protein, and was confirmed by immunoblotting with a specific anti-[Formula: see text]-actinin-4 MAb. This evidence shows, for the first time, that [Formula: see text]-actinin-4 is a protein targeted by AA-I in kidney cells. Our findings strongly suggest an association between [Formula: see text]-actinin-4 and AA-I nephrotoxic activity.

Download Full-text

ISOLATION AND CHARACTERIZATION OF ARISTOLOCHIC ACID - I FROM ARISTOLOCHIA INDICA LINN. LEAVES

INDIAN DRUGS ◽

10.53879/id.50.09.p0052 ◽

2013 ◽

Vol 50 (09) ◽

pp. 52-53

Author(s):

M.B Mulik ◽

◽

K.S. Laddha

Keyword(s):

Aristolochic Acid ◽

Chloroform Extract ◽

Isolation Method ◽

Aristolochic Acids ◽

Isolation And Characterization ◽

Isolated Compound ◽

Anti Fungal ◽

Aristolochic Acid I ◽

Aristolochia Species

Aristolochic Acids (AAs) are major components of plants in Aristolochia species. The plant Aristolochia indica Linn. has diverse biological actions such as analgesic, anti-diabetics, anti-bacterial, anti-fungal and treatment of malaria and fevers. Here, we report a new and simple isolation method of Aristolochic Acid-Ι (AA-I) from leaves of Aristolochia indica Linn. Defatted leaves of A. indica were extracted with chloroform by soxhlation. AA-I was further isolated from the chloroform extract by partitioning with aqueous sodium bicarbonate. AA-I was purified by repetitive washings with methanol and recrystallization in methanol yielded yellow leaflets. The purity of the isolated compound was ascertained by HPLC analysis. structural elucidation of the isolated compound was done by IR, MS and NMR spectral analysis.

Download Full-text

Cytomegalovirus infection of human kidney cells in vitro

Kidney International ◽

10.1038/ki.1991.300 ◽

1991 ◽

Vol 40 (5) ◽

pp. 954-960 ◽

Cited By ~ 16

Author(s):

Jarkko A. Ustinov ◽

Raisa J. Loginov ◽

Pirkko M. Mattila ◽

Virpi K. Nieminen ◽

Jukka I. Suni ◽

...

Keyword(s):

Cytomegalovirus Infection ◽

Human Kidney ◽

Kidney Cells

Download Full-text

Evaluation of biomarkers for in vitro prediction of drug-induced nephrotoxicity in RPTEC/TERT1 cells

Toxicology Research ◽

10.1093/toxres/tfaa005 ◽

2020 ◽

Vol 9 (2) ◽

pp. 91-100 ◽

Cited By ~ 1

Author(s):

Xuan Qiu ◽

Yufa Miao ◽

Xingchao Geng ◽

Xiaobing Zhou ◽

Bo Li

Keyword(s):

Transcriptional Activation ◽

Messenger Rna ◽

Aristolochic Acid ◽

Kidney Injury ◽

Drug Induced ◽

Vitro Assay ◽

Protein Levels ◽

Aristolochic Acid I

Abstract There have been intensive efforts to identify in vivo biomarkers that can be used to monitor drug-induced kidney damage before significant impairment occurs. Kidney injury molecule-1, neutrophil gelatinase-associated lipocalin, clusterin, β2-microglobulin and cystatin C (CysC) have been validated as clinical or preclinical biomarkers in urinary and plasma predictive of acute and chronic kidney injuries and diseases. A high-throughput in vitro assay predictive of nephrotoxicity could potentially be implemented in early drug discovery stage to reduce attrition at later stages of drug development. To assess the potential of these known in vivo biomarkers for in vitro evaluation of drug-induced nephrotoxicity, we selected four nephrotoxic agents (cisplatin, cyclosporin, aristolochic acid I and gentamicin) and detected their effects on the protein levels of nephrotoxic biomarkers in RPTEC/TERT1 cells. The protein levels of clusterin, CysC, GSTπ and TIMP-1 significantly increased in the conditioned media of RPTEC/TERT1 cells treated with cisplatin, cyclosporin, aristolochic acid I and gentamicin. The messenger RNA levels of clusterin, CysC, GSTπ and TIMP-1 also increased in RPTEC/TERT1 cells treated with cisplatin, cyclosporin, aristolochic acid I and gentamicin, indicating that drug-induced upregulation involves transcriptional activation. Taken together, the results clearly demonstrate that among the known in vivo nephrotoxic biomarkers, clusterin, CysC, GSTπ and TIMP-1 can be effectively used as in vitro biomarkers for drug-induced nephrotoxicity in RPTEC/TERT1 cells.

Download Full-text

The Effect of Overexpression of the Enhancer of Zeste Homolog 1 (EZH1) Gene on Aristolochic Acid-Induced Injury in HK-2 Human Kidney Proximal Tubule Cells In Vitro

Medical Science Monitor ◽

10.12659/msm.911611 ◽

2019 ◽

Vol 25 ◽

pp. 801-810 ◽

Cited By ~ 4

Author(s):

Liping Wang ◽

Ning Liu ◽

Xiaoyan Xue ◽

Shujun Zhou

Keyword(s):

Proximal Tubule ◽

Aristolochic Acid ◽

Human Kidney ◽

Proximal Tubule Cells ◽

Kidney Proximal Tubule ◽

Enhancer Of Zeste ◽

Tubule Cells

Download Full-text

Synthesis in vitro of urokinase-like material using polyA(+)RNA from human kidney and from cultured human embryonic kidney cells

Biochemical and Biophysical Research Communications ◽

10.1016/s0006-291x(80)80155-0 ◽

1980 ◽

Vol 97 (1) ◽

pp. 207-215 ◽

Cited By ~ 6

Author(s):

A. Bollen ◽

C. Glineur ◽

A. Herzog

Keyword(s):

Human Kidney ◽

Kidney Cells ◽

Human Embryonic Kidney ◽

Human Embryonic Kidney Cells

Download Full-text

Phytochemical and Biological Investigation of Aristolochia maurorum L.

Zeitschrift für Naturforschung C ◽

10.1515/znc-2006-9-1013 ◽

2006 ◽

Vol 61 (9-10) ◽

pp. 685-691 ◽

Cited By ~ 3

Author(s):

Feras Q. Alali ◽

Khaled Tawaha ◽

Mayadah B. Shehadeh ◽

Suha Telfah

Keyword(s):

Aristolochic Acid ◽

Extraction Methods ◽

Cytotoxic Agents ◽

Flowering Stage ◽

Biological Investigation ◽

Plant Parts ◽

Methanolic Extracts ◽

External Reference ◽

First Time ◽

Aristolochic Acid I

AbstractAristolochia maurorum L. of Jordanian origin has been investigated phytochemically, quantitatively, and biologically. Three atypical alkaloids, namely aristolochic acid I (1), aristolochic acid II (2) and aristolochic acid IIIa (3), have been isolated and identified. Of these known 1-phenanthrenecarboxylic acids, 2 and 3 are reported for the first time from this species. The identified compounds 1-3 were first evaluated biologically as cytotoxic agents against the brine shrimp lethality test (BST), in which compound 1 was found to be the most potent (LC50, 4.9 μg/mL). The antiplatelet activity of the methanolic extracts, the acidic fractions of aerial and root parts, and the identified compounds 1-3 were evaluated using an automatic platelet aggregometer and coagulation tracer (APACT 2). Using external reference standards, and a reverse-phase isocratic method, the distribution of aristolochic acid I and aristolochic acid II in different plant parts of Aristolochia maurorum L. during flowering stage was analyzed by PDA-HPLC. A quantitative comparison between two previously reported extraction methods was also made. Roots were found to be the main storage of aristolochic acid I and aristolochic acid II during flowering stage with about 0.22 and 0.108% (w/w), respectively.

Download Full-text

Prediction of in vivo nephrotoxicity using an in vitro-in silico approach: The case of aristolochic acid I

Toxicology Letters ◽

10.1016/j.toxlet.2015.08.312 ◽

2015 ◽

Vol 238 (2) ◽

pp. S94

Author(s):

R. Abdullah ◽

W. Alhusainy ◽

I.M.C. Rietjens ◽

A. Punt

Keyword(s):

In Silico ◽

Aristolochic Acid ◽

Aristolochic Acid I

Download Full-text

Congener-Specific Polychlorinated Biphenyl–Induced Cell Death in Human Kidney Cells In Vitro: Potential Role of Caspase

International Journal of Toxicology ◽

10.1080/10915810600840859 ◽

2006 ◽

Vol 25 (5) ◽

pp. 341-347 ◽

Cited By ~ 12

Author(s):

Y. Q. Chen ◽

S. De ◽

S. Ghosh ◽

S. K. Dutta

Keyword(s):

Cell Death ◽

Proximal Tubule ◽

Human Kidney ◽

Renal Proximal Tubule ◽

Proximal Tubule Cell ◽

Kidney Cells ◽

Dependent Manner ◽

Caspase Inhibitor ◽

Pcb Congeners

Polychlorinated biphenyls (PCBs) are among the most widespread and persistent pollutants in the global environment. Coplanar and noncoplanar PCBs have been shown to cause congener-specific apoptosis mediated neurotoxicity in rats. Very few, if any, such studies have been reported on human renal cell toxicity. The authors report here caspase-dependent or caspase-independent renal toxicity, as measured by apoptotic death induced by PCBs, depending on the planarity of congeners PCB-77 (coplanar) and PCB-153 (noncoplanar) in human kidney cells (HK2) in vitro. The authors have combined morphological and biological techniques to discover the relevance of apoptosis in renal proximal tubule cell death induced by these two PCB congeners. Treatment with both PCB congeners caused accelerated apoptosis in a time-and concentration-dependent manner. Based on our findings using human kidney (HK2) cells, there was more apoptosis-mediated loss of cell viability by non– ortho-substituted PCB-77 when compared to PCB-153. A significant increase of caspase-3 expression through immunoblot studies showed the involvement of apoptosis by PCB-77 compared to none by PCB-153. The broad-spectrum caspase inhibitor z-VAD-fmk showed increased cell death when treated by PCB-153, but not by PCB-77, confirming that caspase inhibitor induced a switch in the mode of cell death. It is reasonable to assume that apoptotic cell death in the renal proximal tubule cells treated by PCBs may have both caspase-dependent and caspase-independent pathways.

Download Full-text