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Partial least squares discriminant analysis (PLS-DA) score and S-plot of saline control (Ctrl) and cinnabar and reaglar co-administration (C+R) groups based on the serum metabolic profiles for (A) positive (R²X = 0.771, R²Y = 0.989, and Q² = 0.932) and (B) negative (R²X = 0.507, R²Y = 0.944, and Q² = 0.854) ion mode.
Source publication
An-Gong-Niu-Huang Wan (AGNH) is a well-known traditional Chinese medicine (TCM) recipe containing cinnabar (HgS) and realgar (As2S2). However, the application of AGNH is limited by the hepato- and nephrotoxicity of cinnabar and realgar. It should be noted that cinnabar and realgar in AGNH are not used alone, but rather combined with other herbs as...
Citations
... Cerebral ischemia injury can also be alleviated by cinnabar (HgS) and realgar (As 4 S 4 ) containing An-Gong-Niu-Huang Wan (AGNH), and cinnabar and realgar are shown to be essential ingredients in the recipe [13]. Te entire AGNH formulae can also protect against the hepatorenal toxicity produced by cinnabar and realgar alone and reduce Hg and As accumulation in tissues, implying herbal constituents in polyherbo-mineral preparations could ofset the toxic efects of minerals [14,15]. ...
... Chemical reactions may occur due to the mixing of agents, and it is assumed that the various components promote each other to achieve the desired therapeutic effect and reduce toxicity [29]. Studies have confirmed that the toxicity of cinnabar is low and that the herbal medicines in cinnabar formulas can attenuate the damage caused by cinnabar to body systems [29,139,140], which may be related to the absorption, distribution, and excretion of Mercury [28,138]. Similarly, when realgar is used in a formula, the other herbal medicines in the formula can significantly reduce realgar toxicity by reducing blood arsenic levels [29,141]. ...
... Metabolomics studies have confirmed that the other herbs present in AGNH alleviate inflammatory cell infiltration and damage in the liver and kidneys as well as the serum metabolic profile disruptions induced by cinnabar and realgar insults [139,142]. e mechanism may involve protective effects of AGNH's herbal constituents against the accumulation of Hg and As and the hepatorenal toxicity induced by cinnabar and realgar via downregulation of the expression of uptake transporters and upregulation of the expression of efflux transporters in hepatorenal tissues [141]. Similar studies on LSW [143] and Niuhuang Jiedu tablets [144] have also demonstrated that other herbal ingredients in the formulas can reduce the damage caused by realgar to some extent. ...
Materials and methods:
A search was performed for the literature on cinnabar and realgar in PubMed, the Chinese Pharmacopeia, Google, and other sources. The search included studies using single herbs, traditional formulations, or novel dosage forms.
Results:
Cinnabar and cinnabar formulas exhibit good efficacy for sedation, sleep improvement, anxiety alleviation, and brain protection. However, previous studies on neurotransmitters have reached different conclusions, and detailed pharmacological mechanisms are lacking. Realgar and its formulas exert promising antitumor activity through regulation of cell cycle arrest, intrinsic and extrinsic apoptosis, induction of differentiation, autophagy, metabolic reprogramming, matrix metalloproteinase-9 (MMP-9) signaling, and reactive oxygen species (ROS) generation. In addition, realgar can be used to treat a variety of refractory diseases by regulating immunity and exerting antibacterial, antiviral, and other effects. However, the existing pharmacological research on the use of realgar for epidemic prevention is insufficient, and animal experiments and research at the cellular level are lacking. Inappropriate applications of cinnabar and realgar can cause toxicity, including neurotoxicity, liver toxicity, kidney toxicity, and genotoxicity. The toxicological mechanism is complex, and molecular-level research is limited. For clinical applications, theory and clinical experience must be combined to guide scientific and rational drug use and to achieve reduced toxicity and increased efficacy through the use of modern preparation methods or combined drugs. Notably, when cinnabar and realgar are used to treat targeted diseases, these agents have a bidirectional effect of "treatment" and "toxicity" on the central nervous system in pathological and normal states. The pharmacological and toxicological mechanisms need to be elucidated in greater detail in the future. Overall, systematic research is needed to provide a basis for better promotion of the rational use of cinnabar and realgar in the clinic.
Conclusion:
Mineral medicines are multicomponent, multiactivity, and multitargeted substances. The pharmacology and mechanisms of the toxicity and action of realgar and cinnabar are extremely complex. A number of Chinese medicinal preparations of realgar and cinnabar have demonstrated unique efficacy in the treatment of refractory diseases.
... However, poisoning cases caused by overdose or long-term use of realgar-or cinnabar-containing TCMs have been reported occasionally Chang et al., 2018). AGNHP is forbidden in foreign countries because it contains excessive levels of arsenic and mercury (Xia et al., 2018). Since realgar and cinnabar are the essentially active components of AGNHP (Xia et al., 2018;Tsoi et al., 2019), the safety of AGNHP has aroused great concerns among the public. ...
... AGNHP is forbidden in foreign countries because it contains excessive levels of arsenic and mercury (Xia et al., 2018). Since realgar and cinnabar are the essentially active components of AGNHP (Xia et al., 2018;Tsoi et al., 2019), the safety of AGNHP has aroused great concerns among the public. Previous studies reported that AGNHP was protective against cinnabar-and realgar-induced hepatic and renal damage Wang et al., 2021). ...
Realgar- and cinnabar-containing AnGongNiuHuang Pill (AGNHP) is widely used for treating encephalopathy syndrome. However, it raises great safety concerns due to the adverse effects reported by arsenic or mercury poisoning. Although AGNHP has been generally recognized, little is known about the metabolism of arsenic and mercury and their resulting potential health risk in vivo . Thus, comparative pharmacokinetics and urinary excretion of arsenic and mercury were conducted in rats after oral administration of realgar, cinnabar and AGNHP, respectively. The contents of arsenic and mercury in rat blood and urine were determined by hydride-generation atomic fluorescence spectrometry (HG-AFS) after wet digestion. AGNHP significantly reduced the absorption of arsenic in blood and promoted urinary arsenic excretion. Whereas, it increased the blood mercury absorption and reduced urinary mercury excretion. No significant toxicity was observed in the clinical dose range of AGNHP. However, excessive exposure to arsenic and mercury may still pose risks especially by long-term or excessive medication. The results are helpful for the rational clinical applications of realgar- and cinnabar-containing TCMs.
... They were fed freely water and diet, after 1 week of acclimatization, the mice were randomly divided into four groups for the treatment of 0.5% CMC-Na (Control group), Realgar suspended in 0.5% CMC-Na (0.5, 1.0, and 2.0 g/kg), respectively. The doses of Realar were referred to the basis of previously published literature (Miao et al., 2011;Xia et al., 2018). The detailed experiment procedure was shown in Figure 1. ...
Realgar has been used as a type of mineral drug that contains arsenic for thousands of years. Previous studies have shown that Realgar-induced acute kidney injury is associated with abnormal metabolism, but the underlying mechanism is poorly understood. The aim of this study is to investigate the metabolic changes in serum and kidney tissues of mice exposed to Realgar by using a metabolomic approach and explore the molecular mechanisms of acute kidney injury induced by Realgar. Forty mice were randomly divided into four groups: Control group, 0.5-, 1.0, and 2.0 g/kg Realgar group. After 1 week, the body weight and kidney weight of the mice were measured. The serum and kidney samples were used for LC-MS spectroscopic metabolic profiling. Principal component analysis (PCA), correlation analysis, and pathway analysis were used to detect the nephrotoxic effects of Realgar. Body weight decreased significantly in the 2.0 g/kg group, and the kidney weight index also showed a dose-dependent increase in Realgar. The PCA score plot showed the serum and kidney tissue metabolic profile of mice exposed to 2.0 g/kg Realgar separated from the control group, while the lower-doses of 0.5 g/kg and 1.0 g/kg Realgar shown a similar view to the Control group. Thirty-three metabolites and seventeen metabolites were screened and identified in the serum and kidney of mice in a dose-dependent manner. respectively. Correlation analysis showed a strong correlation among these metabolites. Amino acid metabolism, lipid metabolism, glutathione metabolism, and purine metabolism pathways were found to be mainly associated with Realgar nephrotoxicity. This work illustrated the metabolic alterations in Realgar-induced nephrotoxic mice through a metabolomic approach.
... e coordinated action of efflux and uptake transporters regulates the amount of mercuric and arsenical glutathione S-conjugates that transverse out of or return to hepatocytes and renal tubular cells. Our previous studies have shown that the combination of herbal medicine in AGNH can mitigate inflammation and injury in the liver and kidney tissues of mice induced by C + R [13,14]. However, little is known about how AGNH's herbal ingredients act on phase-2 sulfotransferases and phase-3 uptake and efflux transporters induced by C + R. ...
... Since equal amounts (0.056 g) of cinnabar and realgar were contained in per gram AGNH, one group of mice was treated by oral administration of cinnabar (0.14 g/ kg) and realgar (0.14 g/kg) per day for comparison. e details of the dosing regimen were described in our previous report [13,14]. At 1 h after the last dosing, mice were anesthetized by pentobarbital sodium (50 mg/kg) intraperitoneally and sacrificed by cervical dislocation after collection of blood samples from the eyeballs. ...
... Consecutive 28day intragastric administration of C + R merely increased the aspartate aminotransferase (AST) activity as well as the alanine aminotransferase (ALT) activity in liver homogenates. However, it did not significantly influence blood biochemical parameters, suggesting routine serum biochemical markers may not be sensitive indices of hepatorenal impairment induced by C + R [14]. MTs are a family of cysteine-rich intracellular proteins with a well-known metal-binding ability that participate in heavy metal detoxification. ...
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.
... A recent study used UPLC/Q-TOF-MS combined with a pattern recognition approach to identify the detoxicant ingredients of AGNHW. The results revealed that the herbal ingredients of AGNHW could regulate 36 of 41 endogenous metabolites related to the hepatorenal toxicity of cinnabar and realgar [251]. Therefore, metabolomics studies indicate that HLJDD and AGNHW are useful TCM formulae with antioxidant, anti-inflammation and neuroprotective properties for ischemic stroke. ...
Oxidative/nitrosative stress and neuroinflammation are critical pathological processes in cerebral ischemia-reperfusion injury, and their intimate interactions mediate neuronal damage, blood-brain barrier (BBB) damage and hemorrhagic transformation (HT) during ischemic stroke. We review current progress towards understanding the interactions of oxidative/nitrosative stress and inflammatory responses in ischemic brain injury. The interactions between reactive oxygen species (ROS)/reactive nitrogen species (RNS) and innate immune receptors such as TLR2/4, NOD-like receptor, RAGE, and scavenger receptors are crucial pathological mechanisms that amplify brain damage during cerebral ischemic brain injury. Furthermore, we review the current progress of omics and systematic biology approaches for studying complex network regulations related to oxidative/nitrosative stress and inflammation in the pathology of ischemic stroke. Targeting oxidative/nitrosative stress and neuroinflammation could be a promising therapeutic strategy for ischemic stroke treatment.
We then review recent advances in discovering compounds from medicinal herbs with the bioactivities of simultaneously regulating oxidative/nitrosative stress and pro-inflammatory molecules for minimizing ischemic brain injury. These compounds include sesamin, baicalin, salvianolic acid A, 6-paradol, silymarin, apocynin, 3H-1,2-Dithiole-3-thione, (−)-epicatechin, rutin, Dl-3-N-butylphthalide, and naringin. We finally summarize recent developments of the omics and systematic biology approaches for exploring the molecular mechanisms and active compounds of Traditional Chinese Medicine (TCM) formulae with the properties of antioxidant and anti-inflammation for neuroprotection. The comprehensive omics and systematic biology approaches provide powerful tools for exploring therapeutic principles of TCM formulae and developing precision medicine for stroke treatment.
... Xia et al. established an ultrahigh performance liquid chromatography-quadrupole time-of-ight mass spectrometry (UPLC/Q-TOFMS)-based metabolomics approach to reveal the protective role of other herbs in An-gong Niuhuang Wan against the hepatorenal toxicity of cinnabar and realgar. 89 The results showed that the toxicity of the prescription was lower than that of cinnabar and realgar alone. In addition, other TCMs have anti-inammatory and attenuation effects by regulating the metabolism of glycerophospholipid, arachidonic acid, linoleic acid, ether lipid and sphingolipid. ...
Metabolomics is the study of the investigation of small molecules derived from cellular and organism metabolism, which reflects the outcomes of the complex network of biochemical reactions in living systems. As the most recent member of the omics family, there has been notable progress in metabolomics in the last decade, mainly driven by the improvement in mass spectrometry (MS). MS-based metabolomic strategies in modern health and medical science studies provide innovative tools for novel diagnostic and prognostic approaches, as well as an augmented role in drug development, nutrition science, toxicology, and forensic science. In the present review, we not only introduce the application of MS-based metabolomics in the above fields, but also discuss the MS analysis technologies commonly used in metabolomics and the application of metabolomics in precision medicine, and further explore the challenges and perspectives of metabolomics in the field of health and medical science, which are expected to make a little contribution to the better development of metabolomics.
... Cinnabar and realgar are not used alone in AGNHW, but rather combined with different kinds of medicinal herbs as a formula. The medicinal herbs such as Rhizoma Coptidis, Scutellaria Baicalensis, Fructus Gardeniae and Radix Curcumae, in the formula possess antioxidant and anti-inflammatory properties Xia et al., 2018). The combination of the herbs in AGNHW alleviated cinnabar and realgar-induced histopathological alterations and oxidative stress in the liver and kidneys . ...
Realgar and cinnabar are commonly used mineral medicine containing arsenic and mercury in Traditional Chinese Medicine (TCM). Angong Niuhuang Wan (AGNHW) is a representative realgar- and cinnabar-containing TCM formula for treating acute ischemic stroke, but its toxicology and neuropharmacological effects are not well addressed. In this study, we compared the neuropharmacological effects of AGNHW and modified AGNHW in an experimental ischemic stroke rat model. Male SD rats were subjected to 2 h of middle cerebral artery occlusion (MCAO) plus 22 h of reperfusion. Although oral administration of AGNHW for 7 days in the rats increased arsenic level in the blood and liver tissue, there were no significant changes in the arsenic level in kidney, mercury level in the blood, liver and kidney as well as hepatic and renal functions in MCAO rats. AGNHW revealed neuroprotective properties by reducing infarction volume, preserving blood-brain barrier integrity and improving neurological functions against cerebral ischemia-reperfusion injury. Interestingly, removing realgar and/or cinnabar from AGNHW abolished the neuroprotective effects. Meanwhile, AGNHW could scavenge peroxynitrite, down-regulate the expression of p47phox, 3-NT and MMP-9 and up-regulate the expression of ZO-1 and claudin-5 in the ischemic brains, which were abolished by removing realgar and/or cinnabar from AGNHW. Notably, realgar or cinnabar had no neuroprotection when used alone. Taken together, oral administration of AGNHW for one week should be safe for treating ischemic stroke with neuroprotective effects. Realgar and cinnabar are necessary elements with synergetic actions with other herbal materials for the neuroprotective effects of AGNHW against cerebral ischemia-reperfusion injury.
... Using proteomic methods, a chemical probe (i.e., ''bait") of HgS was built and then used to seek and ''hook" the specific targeted protein in nerve cells. The aim of this experiment was to recognize the targeted protein that the main ingredient of cinnabar, HgS, exerted its pharmacological or toxicological effect upon [43]. ...
Traditional Chinese medicine (TCM) is a medical system that has collected and summarized abundant clinical experience over its long history of more than 2000 years. However, the frequent occurrence of TCM-induced adverse reactions has hindered the modernization and internationalization of TCM, while attracting increasing attention from around the world. Unlike chemical drugs and biological agents, the difficulties involved in research on the toxicity and safety of TCM mainly include the complexity of its components and the unpredictability of drug–body interactions. Much of TCM, which has overall therapeutic effects, has the typical mechanisms of multiple components, multiple pathways, and multiple targets. While considering the gradualness and unpredictability of TCM toxicity, the ambiguity of toxicants and safe dosage, and individual differences during long-term TCM administration, we have systematically established key techniques for the toxicity assessment of TCM. These techniques mainly include TCM toxicity discovery in an early phase, based on a combination of drug toxicology genomics and metabolomics; methods to identify dose–toxicity relationships in TCM; and integrated techniques for the exploration of TCM interactions, such as fast-screening tests based on drug-metabolizing enzymes and receptor pathways. In particular, we have developed a new technical system for TCM safety evaluation using molecular toxicology, which has been validated well in research on TCM compatibility contraindication, quality control, and allergen discovery. The application of this key technical platform is introduced here in detail. This application includes model organisms, toxicant biomarkers, a magnetic suspension technique, and the application of network toxicology and computational toxicology in research on the toxicity of Fructus toosendan, Semen cassiae, Polygonum multiflorum, and Fructus psoraleae.
... In this regard, researchers should take mixtures as a whole in intact animals with the traditional oral route of administration to evaluate its action before dissecting the components. For example, HgS and As 4 S 4 containing An-Gong-Niu-Huang Wan protects against the hepatorenal toxicity of HgS and As 4 S 4 when minerals were used alone [101]. ...
Minerals are alchemically processed as Bhasmas in Ayurvedic medicines or as Zuotai in Tibetan medicines. Ayurveda is a knowledge system of longevity and considers the mineral elixir made from “nature” capable of giving humans perpetual life. Herbo-metallic preparations have a long history in the treatment of various diseases in India, China, and around the world. Their disposition, pharmacology, efficacy, and safety require scientific evaluation. This review discusses the Bhasmas in Ayurvedic medicines and Zuotai in Tibetan medicines for their occurrence, bioaccessibility, therapeutic use, pharmacology, toxicity, and research perspectives. A literature search on Mineral, Bhasma, Ayurvedic medicine, Zuotai, Tibetan medicine, and Metals/metalloids from PubMed, Google and other sources was carried out, and the relevant papers on their traditional use, pharmacology, and toxicity were selected and analyzed. Minerals are processed to form Bhasma or Zuotai to alter their physiochemical properties distinguishing them from environmental metals. The metals found in Ayurveda are mainly from the intentional addition in the form of Bhasma or Zuotai. Bhasma and Zuotai are often used in combination with other herbals and/or animal-based products as mixtures. The advanced technologies are now utilized to characterize herbo-metallic preparations as Quality Assurance/Quality Control. The bioaccessibility, absorption, distribution, metabolism, and elimination of herbo-metallic preparations are different from environmental metals. The pharmacological basis of Bhasma in Ayurveda and Zuotai in Tibetan medicines and their interactions with drugs require scientific research. Although the toxic potentials of Bhasma and Zuotai differ from environmental metals, the metal poisoning case reports, especially lead (Pb), mercury (Hg), and arsenic (As) from inappropriate use of traditional medicines, are increasing, and pharmacovigilance is desired. In risk assessment, chemical forms of metals in Bhasma and Zuotai should be considered for their disposition, efficacy, and toxicity.
