Pinellia ternata (Thunb.) Breit. attenuates the allergic airway inflammation of cold asthma via inhibiting the activation of TLR4-medicated NF-kB and NLRP3 signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Pinellia ternata (Thunb.) Breit. (PT) has been demonstrated to be effective against the allergic airway inflammation (AAI) in clinical practices, especially in cold asthma (CA). Until now, the active ingredients, protective effect, and possible mechanism of PT against CA remain unknown. AIM OF THE STUDY: The aim of this investigation was to examine the therapeutic impact and elucidate the underlying mechanism of PT on the AAI of CA. METHODS: The compositions of PT water extract were determined via the UPLC-Q-TOF-MS/MS. The ovalbumin (OVA) and cold-water baths were used to induce CA in female mice. Morphological characteristic observations, expectorant effect, bronchial hyperreactivity (BHR), excessive mucus secretion, and inflammatory factors were used to uncover the treatment effect of PT water extract. In addition, the mucin 5AC (MUC5AC) mRNA and protein levels and the aquaporin 5 (AQP5) mRNA and protein levels were detected via qRT-PCR, immunohistochemistry (IHC), and western blotting. Moreover, the protein expressions associated with the TLR4, NF-κB, and NLRP3 signaling pathway were monitored by western blot analysis. RESULTS: Thirty-eight compounds were identified from PT water extract. PT showed significant therapeutic effects on mice with cold asthma in terms of expectorant activity, histopathological changes, airway inflammation, mucus secretion, and hyperreactivity. PT exhibited good anti-inflammatory effects in vitro and in vivo. The expression levels of MUC5AC mRNA and protein decreased significantly, while AQP5 expression levels increased significantly in the lung tissues of mice after administration with PT as compared to mice induced by CA. Furthermore, the protein expressions of TLR4, p-iκB, p-p65, IL-1ß, IL-18, NLRP3, cleaved caspase-1, and ASC were markedly reduced following PT treatment. CONCLUSIONS: PT attenuated the AAI of CA by modulating Th1- and Th2-type cytokines. PT could inhibit the TLR4-medicated NF-kB signaling pathway and activate the NLRP3 inflammasome to reduce CA. This study provides an alternative therapeutic agent of the AAI of CA after administration with PT.

Processed product (Pinelliae Rhizoma Praeparatum) of Pinellia ternata (Thunb.) Breit. Alleviates the allergic airway inflammation of cold phlegm via regulation of PKC/EGFR/MAPK/PI3K-AKT signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Pinelliae Rhizoma Praeparatum (PRP) is a traditional processed product of Pinellia ternata (Thunb.) Berit., which mainly used for treating cold asthma (CA). However, the mechanism of action of PRP for treating CA have not been fully elucidated. AIM OF THE STUDY: To investigate the core active constituents and the pharmacological mechanism of PRP against CA. MATERIALS AND METHODS: Ovalbumin (OVA) and cold water-induced cold asthma model were established in male mice. The effects of water extract from PRP were evaluated by general morphological observation, expectorant activity, airway hyperresponsiveness, mucus hypersecretion, inflammatory cytokines, etc. Additionally, the mRNA and protein expression of mucin 5AC (MUC5AC) and aquaporin 5 (AQP5) in vivo and in vitro were detected by immunohistochemistry (IHC), qRT-PCR, and western blotting. The mechanisms of action were investigated through network pharmacology and transcriptomic, and validated through western blotting and molecular docking. RESULTS: PRP exhibited a favorable expectorant activity, and significantly reduced the airway inflammation, mucus secretion, and hyperresponsiveness in cold asthma model. It also reduced the levels of IL-4, IL-5, IL-8, and IL-13 in bronchoalveolar lavage fluid (BALF) and IL-4 and total IgE in serum, while obviously increased the levels of IL-10 and IFN-γ in serum for asthmatic mice. Meanwhile, PRP also attenuated the pathological changes and mucus production in cold asthmatic mice. Moreover, the downregulation of MUC5AC and upregulation of AQP 5 were detected by western blotting and qRT-PCR after administration with PRP both in vivo and in vitro. PRP expectedly inhibited the protein expression of PKC-α, SRC, p-EGFR, p-ERK1/2, p-JNK, p-p38, p-PI3K, and p-Akt levels in vivo. CONCLUSIONS: These combined data showed that PRP suppressed the allergic airway inflammation of CA by regulating the balance of Th1 and Th2 cytokines and the possible involvement of the PKC/EGFR/MAPK/PI3K-Akt signaling pathway. Pentadecanoic acid, licochalcone A, ß-sitosterol, etc. were considered as main active ingredients of PRP against CA. This study provides a novel perspective of the classical herbal processed product PRP in the treatment of CA.

Effect of Mashao Pingchuan Decoction on Traditional Chinese Medicine Symptoms and Quality of Life in Patients with Cold Asthma of Bronchial Asthma / 中国实验方剂学杂志

Objective:To evaluate the effect of Mashao Pingchuan decoction on traditional Chinese medicine <bold>（</bold>TCM<bold>）</bold> symptoms， quality of life， peripheral blood eosinophils （Eos） and serum inflammatory factors interleukin-6 （IL-6） and interleukin-8 （IL-8） in patients with cold asthma syndrome of bronchial asthma. Method:A total of 67 patients with cold asthma who attended the Respiratory Clinic of Anhui Provincial Hospital of TCM from January 2018 to December 2019 were selected and randomly divided into a control group and an observation group. The control group was given basic treatments such as budesonide formoterol powder inhalation， and the observation group was given Mashao Pingchuan decoction on the basis of the control group. Both groups intervened for 7 consecutive days. Observe and record the general condition， TCM symptom score， asthma control questionnaire （ACQ）-7 score， Marks-asthma quality of life questionnaire （Marks-AQLQ） score of the two groups of asthma patients， the peripheral blood Eos count was measured by hematology analyzer， and the serum IL-6 and IL-8 levels of the subjects were measured by enzyme-linked immunosorbent assay （ELISA）. Result:The total effective rate of TCM symptoms in the two groups was 100%（30/30）， and the effect in the observation group was more obvious （<italic>Z</italic>=-2.169，<italic>P</italic><0.05<italic>）</italic>. After treatment， the scores of TCM symptoms in the two groups were significantly lower than those before treatment （<italic>P</italic><0.01）. The scores of phlegm in the throat， expectoration， cough， and chills in the observation group were lower than those in the control group （<italic>P</italic><0.05）. After treatment， the ACQ-7 scores and Marks-AQLQ scores of the two groups were significantly lower than before treatment （<italic>P</italic><0.01）. There was no statistically significant difference between the observation group and the ACQ-7 scores and Marks-AQLQ scores. After treatment， the peripheral blood Eos counts and serum IL-6 and IL-8 levels in the two groups were significantly lower than before treatment （<italic>P</italic><0.01）. Peripheral blood Eos count and serum IL-6 and IL-8 contents in observation group were lower than those in control group （<italic>P</italic><0.05）. Conclusion:Mashao Pingchuan decoction combined with budesonide formoterol powder inhalation can effectively improve the clinical effectiveness of asthma （cold asthma）， improve the symptoms of asthma in TCM， ACQ-7 score， Marks-AQLQ score， peripheral blood Eos count， serum inflammatory factor content.

Mechanism of processing-compatibility of ginger in treating cold asthma rat by gas chromatography-mass spectrometry based on metabolomics / 中草药

Objective: To investigate the effects of processing and compatibility of ginger for the treatment of cold asthma rats at the metabolomics level by gas chromatography-mass spectrometry (GC-MS). Methods: The rats of cold asthma were established by ovalbumin (OVA) + ice water bath. The rats divided randomly into the control group, model group, Linggan Wuwei (fresh/dry/stir- frying) Jiangxin Decoction group and the positive drug Guilong Kechuanning group. The pathological changes of lung tissue were observed by HE staining; The inflammatory cell count in BALF and the content of IgE, IL-4 and IFN-γ in serum were determined. GC-MS was used to conduct the non-targeted metabolomics study to search the serum and urine related differential metabolites in rats with cold asthma, and MetaboAnalyst was used to construct related metabolic pathways. Results: The results showed that Linggan Wuwei (fresh/dry/stir-frying) Jiangxin Decoction improved the pathological changes of rat lung tissue, significantly reduced the BALF inflammatory cell count in BALF and IgE and IL-4 levels in serum, and increased IFN-γ levels. Compared with the control group, 37 differential metabolites (15 in serum and 22 in urine) were screened in cold asthma rats. And seven metabolic pathways involving energy metabolism, oxidative stress may be associated with cold asthma by Metaboanalysis pathway analysis. The overall metabolic profile of the cold asthma rats with the intervention of Linggan Wuwei (fresh/dry/stir-frying) Jiangxin Decoction tended to normal levels. The effect of Linggan Wuwei (dry) Jiangxin Decoction on cold asthma was better than Linggan Wuwei (fresh/stir-frying) Jiangxin Decoction. Conclusion: It is more reasonable to process ginger into dried ginger in Linggan Wuwei Jiangxin Decoction against cold asthma. Ginger processing-compatibility may play a therapeutic role in cold asthma rats by regulating energy metabolism and oxidative stress.

The Study of Dried Ginger and Linggan Wuwei Jiangxin Decoction Treatment of Cold Asthma Rats Using GC-MS Based Metabolomics.

Dried ginger is the monarch drug in Linggan Wuwei Jiangxin (LGWWJX) decoction, which is used to treat cold asthma. The purpose of this study was to investigate and compare the effects of dried ginger and LGWWJX decoction for treatment of cold asthma rats at the metabolomics level using gas chromatography-mass spectrometry (GC-MS). OVA and ice water-induced cold asthma were induced in SD rats. The effects of dried ginger and LGWWJX decoction were evaluated by general morphological observation, hematoxylin and eosin staining, inflammatory cell count, IgE, IL-4, IFN-γ quantitation, and visceral index. GC-MS-based metabolomics was performed and analyzed using multivariate statistical analysis. Biomarker identification, pathway analysis, correlations between identified biomarker, and efficacy indices were performed. The results showed that dried ginger and LGWWJX decoction had obvious effects on cold asthma rats. Thirty-seven metabolites (15 in serum and 22 in urine) associated with cold asthma were identified. These metabolites were mainly carbohydrates, fatty acids and their products, organic acids, and others. Seven pathways were identified by MetaboAnalyst 4.0 metabolic pathway analysis. After intervention with dried ginger and LGWWJX decoction, the majority of altered metabolites and metabolic pathways returned to control levels. LGWWJX decoction regulated more metabolites of carbohydrates and fatty acids, which contribute to energy metabolism and oxidative stress in cold asthma, than dried ginger. We concluded that dried ginger and LGWWJX decoction both were effective for treatment of cold asthma. LGWWJX decoction was more effective than dried ginger for treatment of cold asthma. This study evaluated the effects of dried ginger and LGWWJX decoction on cold asthma at the metabolomics level. It provides a reference for the research on the compatibility of Chinese Medicine.