Determination of the effect of Pinellia ternata (Thunb.) Breit. on nervous system development by proteomics
Graphical abstract
Introduction
Pinelliae rhizoma (Banxia in Chinese) is the dried root of Pinellia ternata (Thunb.) Breit.; the first recorded use in Shen Nong Ben Cao Jing ( a classical work of traditional Chinese medicine [TCM]) was approximately 2000 years ago. It is a commonly used Chinese medicinal herb for the treatment of cough, phlegm, vomiting, and cancer (Kao et al., 2012, Wu et al., 2011). Pharmacological studies have shown that BX has suitable antiemetic properties for the management of severe pregnant vomiting (Grobetal et al., 2014, Kurata et al., 1998). However, raw BX has been reported as an irritant, teratogen, carcinogen, mutagen, and, in particular, are productive toxin (Kao et al., 2012, Wu et al., 2011). It is reported that pregnant rats exposed to high doses of BX extract (0.2 and 2 g/kg) may induce fetal abnormalities (Shin et al., 2007). Our previous study also indicated that a suspension of raw BX powder (6 g/kg) caused embryotoxicity in mice (Xu et al., 2013). Subsequently, we confirmed that raw BX suspension (2.275 g/kg) may affect fetal body length (Xu et al., 2017). Nevertheless, the mechanism of raw BX-induced toxicity is still not fully understood.
As Chinese medical herbs contain multiple chemical components and act on multiple targets (Yu et al., 2012), it is difficult to elucidate the mechanism of herbal toxicity using only conventional research methods such as segment II teratology and histological analyses. For the past few years, high-throughput methods such as proteomics and metabolomics with bioinformatics have emerged as effective approaches to extend drug safety knowledge (Ahuja and Sharma, 2014, Robinson and Piersma, 2013). We previously used a metabolomic approach to evaluate the changes of endogenous metabolites in the placental and amniotic fluid of pregnant rats exposed to BX and 20 significantly different metabolic perturbations were identified (Xie et al., 2016).
Proteomics is a systematic approach for performing proteins studies on a large-scale. It is also useful in revealing the potential targets or biomarkers by studying the targets of pharmaceutical products and analyzing major components of physiological pathways (Suo et al., 2016). The role of proteomics in TCM research has increased insignificance and its use in the study of TCM has experienced a steady increase over many years (Fan et al., 2017, Lao et al., 2014, Liu and Guo, 2011). Among the many proteomics technologies, iTRAQ has become popular for protein identification and quantification owing to its sensitivity, accuracy, and high throughput (Wiese et al., 2007). Herein, an iTRAQ-based proteomic method was used to explore the mechanisms of raw BX-induced fetal toxicity in mice.
Section snippets
Plant material and preparation
BX (batch number 20140210), the Chinese medicinal herb was purchased from Anhui Fengyuan Tongling Herbal Pieces Co., Ltd and authenticated by Professor Shengjin Liu (Nanjing University of Chinese Medicine). The dried raw BX were pulverized and sieved through a 100-mesh sieve in order to prepare a powder. A suspension of 0.17 g/mL BX was prepared by the addition of 51 g BX powder into 300 mL water.
Drugs, animal, groups, and treatments
ICR mice of 10 weeks old were obtained from Vital River Laboratory Animal Technology Co., Ltd
Proteomic expression patterns in gastrulae of BX-treated mice
A shotgun quantitation approach based on iTRAQ was used to obtain an overall summary of proteomic changes in the BX group compared with the control group. A total of 1245 DAPs were identified with < 1% FDR in two biological replicates; 583 protein abundance changes were confidently assessed using at least two unique peptides. When the fold-change threshold cutoff was set at 1.5-fold for increased accumulation and < 0.67-fold for decreased accumulation, 153 proteins with differential
Discussion
Raw BX is a typical herbal medicine, which has been widely used in Chinese clinical practice for thousands of years. However, previous reports showed that BX exerted significant toxicity in pregnant mice and embryos (Grobetal et al., 2014). Shin (2007) revealed that BX might cause fetal abnormalities by influencing the expression of genes related to antioxidants, growth factors, apoptosis, and tumors. Metabolomic studies revealed the mechanisms of BX cardio-toxicity were the inhibition of the
Conclusions
In summary, a shotgun quantitation approach based on iTRAQ was applied to assess protein expression changes caused by BX in mice gastrulae. A total of 1245 proteins were identified with < 1% FDR, and 583 protein abundance changes were confidently assessed. Moreover, 153 proteins identified in BX-treated samples showed significant differences in abundance. Among the DAPs, the functions of 37 proteins were predominantly related to nervous system development, including brain development and neuron
Acknowledgments
This study was financially supported by the National Natural Science Foundation of China (Grant number 81303299).
Conflict of interest
All the authors declare that there are no conflicts of interest.
References (45)
- et al.
Antiviral effects of Jinxin oral liquid against respiratory syncytial virus infection in the BALB/c mice model
J. Ethnopharmacol.
(2015) - et al.
An integrated shotgun proteomics and bioinformatics approach for analysis of brain proteins from MCAO model using serial affinity chromatograph with four active ingredients from Shengmai preparations as ligands
Neurochem. Int.
(2017) - et al.
Twisted gastrulation mutation suppresses skeletal defect phenotypes in Crossveinless 2 mutant mice
Mech. Dev.
(2008) - et al.
Osthole inhibits the invasive ability of human lung adenocarcinoma cells via suppression of NF-kappaB-mediated matrix metalloproteinase-9 expression
Toxicol. Appl. Pharmacol.
(2012) - et al.
Application of proteomics to determine the mechanism of action of traditional Chinese medicine remedies
J. Ethnopharmacol.
(2014) - et al.
Gu-Ben-Fang-Xiao decoction attenuates sustained airway inflammation by suppressing ER stress response in a murine asthma remission model of respiratory syncytial virus infection
J. Ethnopharmacol.
(2016) Nerve ending "signal" proteins GAP-43, MARCKS, and BASP1
Int. Rev. Cytol.
(2005)- et al.
The BASP1 family of myristoylated proteins abundant in axonal termini. Primary structure analysis and physico-chemical properties
Biochimie
(1997) - et al.
Hippocampal cholinergic neurostimulating peptides (HCNP)
Progress. Neurobiol.
(2000) - et al.
The mammalian twisted gastrulation gene functions in foregut and craniofacial development
Dev. Biol.
(2004)
Twisted gastrulation modulates bone morphogenetic protein-induced collagen II and X expression in chondrocytes in vitro and in vivo
J. Biol. Chem.
Effect of Pinellia ternata extract on fetal development of rats
Reprod. Toxicol.
Diazepam binding inhibitor overexpression in mice causes hydrocephalus, decreases plasticity in excitatory synapses and impairs hippocampus-dependent learning
Mol. Cell Neurosci.
Structural characterization of twisted gastrulation provides insights into opposing functions on the BMP signalling pathway
Matrix Biol.
Involvement of the mouse Prp19 gene in neuronal/astroglial cell fate decisions
J. Biol. Chem.
Sedative, hypnotic and anticonvulsant activities of the ethanol fraction from Rhizoma Pinelliae Praeparatum
J. Ethnopharmacol.
Effects of Pinellia ternata (Thunb.) Berit. on the metabolomic profiles of placenta and amniotic fluid in pregnant rats
J. Ethnopharmacol.
Drug safety testing paradigm, current progress and future challenges: an overview
J. Appl. Toxicol.
Loss of profilin-1 expression enhances breast cancer cell motility by Ena/VASP proteins
J. Cell Physiol.
Spinal axon regeneration evoked by replacing two growth cone proteins in adult neurons
Nat. Neurosci.
Purkinje cell maturation participates in the control of oligodendrocyte differentiation: role of sonic hedgehog and vitronectin
PLoS One
Intrinsic neuronal determinants locally regulate extrasynaptic and synaptic growth at the adult neuromuscular junction
J. Cell Biol.
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2023, PhytomedicineCitation Excerpt :Banxia possesses various pharmacological and biological activities, such as antioxidant, antitussive, antiemetic, antitumor, antibacterial, and sedative-hypnotic (Li et al., 2016). The irritating and toxic components of Pinellia ternate are the toxic needle crystals with a special crystalline form, which have strong inflammatory effects and irritation to the local mucous membrane and neurotoxicity at the dose of 2.275 g/kg/day for mice, similar to 15 g/kg/day for human and slightly higher than the recommended dose of pharmacopoeia (Xu et al., 2018). The typhonium giganteum lectin (TGL) (50 µg/ml) had a substantial inflammatory effect and elevated levels of inflammatory factors (TNF-α, IL-1β) (Yanqiu et al.).