Aristolochic Acid Induces Chronic Kidney Disease in ACE Knockout Mice

Jia‑Ping Wu


Background: Aristolochic acid I (AAI) is an extract from Chinese herbs that causes progressive interstitial nephritis. The aim of this research is to know whether chymases play the crucial role in AAI‑induced nephropathy. Methods: The mice were treated with AAI via intraperitoneal injection and the accumulated AAI dosages are 30 mg/kg of body weight for two, four, six, and eight weeks. The animals were sacrificed after another two or four weeks for nephropathy development. Collection of blood, urine, and kidney samples for the further biochemical analysis, hematoxylin–eosin (H and E) and Masson’s trichrome stained to detected pathologic, and MMP2 and MMP9 activity assays. Results: After the treatment of AAI, of the mice, their body weights were decreased (P < 0.01), and concentration of creatinine and blood urea nitrogen (BUN) in serum (P < 0.01) and urine collection were increased (P < 0.01). In the renal tissue sections, high amount of inflammatory cells were found by H and E stain, and increased fibrosis in renal interstitial tissue were observed by Masson’s trichrome stain. In mice kidney tissue, significantly increased chymase activity after treatment of AAI was found (P < 0.01), but ACE activity did not show significant changes. In ACE KO mice, increased MMP2 and decreased MMP9 activity were found in the AAI‑treated mice compared with AAI‑untreated control (P < 0.01). Conclusions: Moreover, it was also observed that the deficiency of ACE would accelerate the disease development of AAI‑induced nephropathy. These results may help to know more information about the role of AAI‑induced chronic kidney disease and can be applied in developing new drug targets for nephropathy.


Aristolochic acid I; chronic kidney disease; chymases; peptidyl-dipeptidase A; renal insufficiency

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Li W, Zhang S. Risk factors of parathyroid dysfunction in elderly

patients with chronic kidney disease undergoing hemodialysis.

Adv Clin Exp Med 2015;24:1007‑12.

Kim SM, Kim KM, Kwon SK, Kim HY.

Erythropoiesis‑stimulating agents and anemia in patients

with non‑dialytic chronic kidney disease. J Korean Med Sci


Hsiao FC, Tung YC, Chou SH, Wu LS, Lin CP, Wang CL,

et al. Fixed‑dose combinations of renin‑angiotensin system

inhibitors and calcium channel blockers in the treatment of

hypertension: A comparison of angiotensin receptor blockers and

angiotensin‑converting enzyme inhibitors. Medicine (Baltimore)

;94:e2355. doi: 10.1097/MD.0000000000002355.

Aragão DS, de Andrade MC, Ebihara F, Watanabe IK,

Magalhães DC, Juliano MA, et al. Serine proteases as candidates

for proteolytic processing of angiotensin‑I converting enzyme.

Int J Biol Macromol 2015;72:673‑9.

Wang R, Chen J, Zhang Z, Cen Y. Role of chymase in the local

renin‑angiotensin system in keloids: Inhibition of chymase may

be an effective therapeutic approach to treat keloids. Drug Des

Devel Ther 2015;9:4979‑88.

Park S, Bivona BJ, Ford SM Jr, Xu S, Kobori H, de Garavilla L,

et al. Direct evidence for intrarenal chymase‑dependent

angiotensin II formation on the diabetic renal microvasculature.

Hypertension 2013;6:465‑71.

DI Carlo A. Matrix metalloproteinase‑2 and ‑9 and tissue

inhibitor of metalloproteinase‑1 and ‑2 in sera and urine of

patients with renal carcinoma. Oncol Lett 2014;7:621‑6.

Bunel V, Antoine MH, Stévigny C, Nortier J, Duez P. New

in vitro insights on a cell death pathway induced by magnolol

and honokiol in aristolochic acid tubulotoxicity. Food Chem

Toxicol 2016;87:77‑87.

Wang K, Feng C, Li C, Yao J, Xie X, Gong L, et al. Baicalin

protects mice from aristolochic acid i‑induced kidney injury by

induction of CYP1A through the aromatic hydrocarbon receptor.

Int J Mol Sci 2015;16:16454‑68.

Feng C, Xie X, Wu M, Li C, Gao M, Liu M, et al. Tanshinone

I protects mice from aristolochic acid I‑induced kidney injury by

induction of CYP1A. Environ Toxicol Pharmacol 2013;36:850‑7.

Xing G, Qi X, Chen M, Wu Y, Yao J, Gong L, et al. Comparison

of the mutagenicity of aristolochic acid I and aristolochic

acid II in the gpt delta transgenic mouse kidney. Mutat Res


Declèves AÉ, Jadot I, Colombaro V, Martin B, Voisin V,

Nortier J, et al. Protective effect of nitric oxide in aristolochic

acid‑induced toxic acute kidney injury: An old friend with new

assets. Exp Physiol 2016;10:193‑206.

Nortier J, Pozdzik A, Roumeguere T, Vanher weghem JL.

[Aristolochic acid nephropathy (“Chinese herb nephropathy”)].

Nephrol Ther 2015;11:574‑88.

Zeng Y, Yang X, Wang J, Fan J, Kong Q, Yu X. Aristolochic

acid I induced autophagy extenuates cell apoptosis via ERK

/2 pathway in renal tubular epithelial cells. PLoS One

;7:e30312. doi: 10.1371/journal.pone. 0030312.

Kwak DH, Lee S. Aristolochic Acid I causes testis toxicity by

inhibiting Akt and ERK1/2 phosphorylation. Chem Res Toxicol


Wu JP, Ho TJ, Tsai CC, Yeh YL, Lin CC, Lin KH, et al.

Hepatoprotective effects of traditional chinese medicine on

liver fibrosis from ethanol administration following partial

hepatectomy. Chin J Physiol 2015;58:393‑403.

Tsai CC, Wu JP, Lin YM, Yeh YL, Ho TJ, Kuo CH, et al. The

effect of elephantopus scaber L. on liver regeneration after

partial hepatectomy. Evid Based Complement Alternat Med

;2013:369180. doi: 10.1155/2013/369180.

McDonald JE, Padmanabhan N, Petrie MC, Hillier C,

Connell JM, McMurray JJ. Vasoconstrictor effect of the

angiotensin‑converting enzyme‑resistant, chymase‑specific

substrate [Pro (11) (D)‑Ala (12)] angiotensin I in human

dorsal hand veins: In vivo demonstration of non‑ace

production of angiotensin II in humans. Circulation


Vass DG, Shrestha B, Haylor J, Hughes J, Marson L.

Inflammatory lymphangiogenesis in a rat transplant model

of interstitial fibrosis and tubular atrophy. Transpl Int


Freise C, Kim KY, Querfeld U. A lindera obtusiloba extract

blocks calcium‑/phosphate‑induced transdifferentiation and

calcification of vascular smooth muscle cells and interferes with

matrix metalloproteinase‑2 and metalloproteinase‑9 and NF‑κB.

Evid Based Complement Alternat Med 2015;2015:679238. doi:


Bojic S, Kotur‑Stevuljevic J, Kalezic N, Stevanovic P,

Jelic‑Ivanovic Z, Bilanovic D, et al. Diagnostic value of

matrix metalloproteinase‑9 and tissue inhibitor of matrix

metalloproteinase‑1 in sepsis‑associated acute kidney injury.

Tohoku J Exp Med 2015;237:103‑9.

Li H, Liu J, Wang W, Zhang Z, Li D, Lin K, et al. Matrix

metalloproteinase 9 and vasodilator‑stimulated phosphoprotein

related to acute kidney injury in severe acute pancreatitis rats.

Dig Dis Sci 2015;60:3647‑55.

DuBose JJ, Azizzadeh A. Utilization of a tubularized cormatrix

extracellular matrix for repair of an arteriovenous fistula

aneurysm. Ann Vasc Surg 2015;29:366.e1‑4.

Bai Y, Lu H, Hu L, Hong D, Ding L, Chen B. Effect of sedum

sarmentosum BUNGE extract on aristolochic acid‑induced renal

tubular epithelial cell injury. J Pharmacol Sci 2014;124:445‑56.