The Herbal Constituents in An-Gong-Niu-Huang Wan (AGNH) Protect against Cinnabar- and Realgar-Induced Hepatorenal Toxicity and Accumulations of Mercury and Arsenic in Mice.

Songsong Wang,Xiao Xiao,Ao Li,Peng Li,Jie Liu

doi:10.1155/2021/5566078

Abstract

An-Gong-Niu-Huang Wan (AGNH) has been a well-known cinnabar- and realgar-containing compound recipe for cerebral diseases. Unfortunately, its clinical practice is often restrained by the specific hepatorenal toxicity of cinnabar and realgar (C + R). In previous research studies, we have found that the antioxidative and anti-inflammatory effects of its herbal constituents could mitigate the risks from the toxicity. The underlying detoxification mechanisms are still unsolved. The present study investigated the protective effects of AGNH's herbal constituents on hepatorenal injury induced by C + R. For the mice treated with C + R, the increased expression levels of sensitive biomarkers of metal exposure and hepatorenal toxicity, including metallothionein (MT) in both hepatorenal tissues and kidney induced molecule-1 (KIM-1) in the kidney, were simultaneously reduced when C + R coadministered with other herbal medicines. In addition, the contents of trivalent As (AsIII), pentavalent As (Asv), and mercury (Hg) in hepatorenal tissues of mice were also significantly reduced benefiting from the herbal constituents in AGNH. Further mechanism studies showed that the herbal constituents in AGNH could downregulate the expressions of uptake transporters (AQP9 and OAT1) and upregulate the expressions of efflux transporters (P-gp, MRP2, and MRP4) in mice intoxicated by C + R. Our results suggested that AGNH's herbal constituents protect the body against C + R-induced hepatorenal toxicity and accumulations of Hg and As, which could be associated with the reestablishment of heavy metal homeostasis and the detoxification system.

Highlights

An-Gong-Niu-Huang Wan (AGNH) has been a renowned compound recipe for treating cerebral diseases [1]
Immunohistochemical analysis showed both hepatocytes and renal proximal tubular epithelial cells (RPTECs) stained strongly positive for MT-1 when mice were treated with cinnabar combined with realgar for 28 days (Figures 1(a) and 1(b)), whereas positive staining for MT-1 was weak in the sections of hepatorenal tissues in mice treated with saline or AGNH
Results obtained from Western Blotting (WB) analysis revealed that MT-1 expression levels in hepatorenal tissues of mice were unchanged between groups of AGNH and saline (Figures 1(d) and 1(e)), but combined administration of C + R exerted a significant increasing effect on MT-1 protein levels in hepatorenal tissues of mice

Summary

Introduction

An-Gong-Niu-Huang Wan (AGNH) has been a renowned compound recipe for treating cerebral diseases [1]. Mineral medicinal materials cinnabar and realgar (C + R) (with 96% of HgS and 90% of As4S4) are contained, accounting for 12.5% by weight in the formula [2]. It is evident that AGNH contains multiple herbal ingredients, which are deemed to function in delivering drugs to target tissues as well as eliminating the harmful influences of the metallic ingredients including mercury (Hg) and arsenic (As) [6]. As a family of phase-2 detoxification enzymes in vivo, glutathione S-transferases (GST) could catalyze the conjugation of reduced glutathione (GSH) to mercuric and arsenical species [7, 8]. E generated Hg- and As- glutathione S-conjugates were transported across the canalicular membrane into the bile by the basolateral efflux transporters, such as P-glycoprotein (P-gp) and members of the multidrug resistanceassociated protein family (MRP) [9]. As a family of phase-2 detoxification enzymes in vivo, glutathione S-transferases (GST) could catalyze the conjugation of reduced glutathione (GSH) to mercuric and arsenical species [7, 8]. e generated Hg- and As- glutathione S-conjugates were transported across the canalicular membrane into the bile by the basolateral efflux transporters, such as P-glycoprotein (P-gp) and members of the multidrug resistanceassociated protein family (MRP) [9]. ese glutathione

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