Objective:

2

To investigate the protective effects of stir-fried

Semen Armeniacae Amarum

(SAA) against aristolochic acid Ⅰ (AAⅠ)-induced nephrotoxicity and DNA adducts and elucidate the underlying mechanism involved for ensuring the safe use of

Asari Radix

et Rhizoma.

Methods:

2

In vitro

HEK293T cells overexpressing Flag-tagged multidrug resistance-associated

protein 3 (MRP3) were constructed by Lentiviral transduction

and inhibitory effect of top 10 common pairs of medicinal herbs with

Asari Radix

et Rhizoma in clinic on MRP3 activity was verified using a self-constructed fluorescence screening system. The mRNA

protein expressions

and enzyme activity levels of NAD(P)H quinone dehydrogenase 1 (NQO1) and cytochrome P450 1A2 (CYP1A2) were measured in differentiated HepaRG cells. Hepatocyte toxicity after inhibition of AAⅠ metabolite transport was detected using cell counting kit-8 assay.

In vivo

C57BL/6 mice were randomly divided into 5 groups according to a random number table

including: control (1% sodium bicarbonate)

AAⅠ (10 mg/kg)

stir-fried SAA (1.75 g/kg) and AAⅠ + stir-fried SAA (1.75 and 8.75 g/kg) groups

6 mice in each group. After 7 days of continuous gavage administration

liver and kidney damages were assessed

and the protein expressions and enzyme activity of liver metabolic enzymes NQO1 and CYP1A2 were determined simultaneously.

Results:

2

In vivo

combination of 1.75 g/kg SAA and 10 mg/kg AAⅠ suppressed AAⅠ-induced nephrotoxicity and reduced dA-ALI formation by 26.7%

and these detoxification effects in a dose-dependent manner (

P

<

0.01). Mechanistically

SAA inhibited MRP3 transport

in vitro

downregulated NQO1 expression

in vivo

increased CYP1A2 expression and enzymatic activity

in vitro

and

in vivo

respectively (

P

<

0.05 or

P

<

0.01). Notably

SAA also reduced AAⅠ-induced hepatotoxicity throughout the detoxification process

as indicated by a 41.3% reduction in the number of liver adducts (

P

<

0.01).

Conclusions:

2

Stir-fried SAA is a novel drug candidate for the suppression of AAⅠ-induced liver and kidney damages. The protective mechanism may be closely related to the regulation of transporters and metabolic enzymes.