Pinelliae Rhizoma Praeparatum Research Articles

UPLC-Q-Exactive/MS based analysis explore the correlation between components variations and anti-influenza virus effect of four quantified extracts of Chaihu Guizhi decoction

Ethnopharmacological relevanceChaihu Guizhi decoction (CGD) is a classic Traditional Chinese Medicine (TCM) prescription for the treatment of influenza and fever, composes of Bupleuri Radix (Chaihu), Cinnamomi Ramulus (Guizhi), Scutellariae Radix (Huangqin), Codonopsis Radix (Dangshen), Glycyrrhizae Radix Et Rhizoma Praeparata Cum Melle (Zhigancao), Pinelliae Rhizoma Praeparatum (Fabanxia), Zingiberis Rhizoma Recens (Shengjiang), Paeoniae Radix Alba (Baishao) and Jujubae Fructus (Dazao) in the ratio of 12:4.5:4.5:4.5:3:6:4.5:4.5:4. The efficacy of TCM, if there are differences, depends on the different extraction methods and extracted components. Aim of the studyThis study was to evaluate the anti-influenza virus effect of CGD extracts with different extraction methods, analyze the components and explore their correlation. Materials and methodsCGD were prepared with four extraction methods respectively, the traditional decoction (TD), two steps alcohol-water extraction (AWE), alcohol reflux extraction (AE) and water reflux extraction (WE). Based on the influenza mouse model, the efficacy of anti-influenza virus in vivo of the four CGD extracts were evaluated with the therapeutic index of body weight, rectal temperature, lung index, thymus index and lung viral load of mice. The chemical components in four CGD extracts, and compounds absorbed in rats blood with prototypes or metabolites were identified by UPLC-Q-Exactive/MS. The partial least squares (PLS) method was used to explore the correlation between the components variation in CGD extracts and the comprehensive efficacy index. The potential effective components were further accessed by molecular docking. ResultsComparing with the other three extracts, AWE has the best anti-influenza effect. It could ameliorate the symptoms caused by influenza virus infection in mice, increase body weight and rectal temperature, reduce the lung index and virus load in lung tissue. 129, 144, 140 and 129 components were identified from TD, AWE, AE, and WE respectively. The identified components were mainly including flavonoids, terpenoids, organic acids, phenylpropanoids, amino acids, nucleosides, phenols, alkaloids, etc. 43 prototypes and 49 metabolites of CGD were detected in rat plasma after oral administration. Seven components, cinnamaldehyde, wogonoside, baicalin, baicalein, gallic acid, oroxylinA-7-O-glucuronide and coumarin, showed significant correlation with anti-influenza effects, all of which had good binding activity with NA, IL-6, STAT3, AKT1, EGFR and TNF. ConclusionTwo steps alcohol-water extraction was optimal for CGD preparation. Cinnamaldehyde, wogonoside, oroxylinA-7-O-glucuronide, coumarin, gallic acid, baicalein and baicalin play a certain essential role in anti-influenza effects and may be taken as a potential maker compounds for quality evaluation of CGD.

Effect of lime water processing of Pinelliae Rhizoma Praeparatum on toxic component lectin protein

The present study investigated the effect of immersion in the excipient lime water on the toxic component lectin protein and explained the scientific connotation of lime water detoxication during the processing of Pinelliae Rhizoma Praeparatum. Western blot was used to investigate the effects of immersion in lime water with different pH(pH 10, 11, and 12.4), saturated sodium hydroxide, and sodium bicarbonate solution on the content of lectin protein. The protein compositions of the supernatant and the precipitate after immersing lectin protein in lime water of different pH were determined by the SDS-PAGE method combined with the silver staining technique. The MALDI-TOF-MS/MS technique was used to detect the molecular weight distribution of peptide fragments in the supernatant and precipitate after immersing lectin protein in lime water of different pH, and circular dichroism spectroscopy was used to detect the ratio changes in the secondary structure of lectin protein during the immersion. The results showed that immersion in lime water at pH>12 and saturated sodium hydroxide solution could significantly reduce the content of lectin protein, while immersion in lime water at pH<12 and sodium bicarbonate solution had no significant effect on lectin protein content. The corresponding lectin protein bands and molecular ion peaks were not detected at the 12 kDa position in the supernatant and precipitate after immersing the lectin protein in lime water at pH>12, which was attributed to the fact that lime water immersion at pH>12 could significantly change the ratio of the secondary structure of lectin protein, resulting in irreversible denaturation, while lime water immersion at pH<12 did not change the ratio of the secondary structure of lectin protein. Therefore, pH>12 was the key condition for the detoxication of lime water during the processing of Pinelliae Rhizoma Praeparatum. Lime water immersion at pH>12 could cause irreversible denaturation of lectin protein, resulting in a significant decrease in the inflammatory toxicity of Pinelliae Rhizoma Praeparatum, which played a key role in detoxification.

Preparation of monoclonal antibodies against Pinellia ternata lectin protein and establishment of double-antibody sandwich ELISA

To determine the content of endogenous toxic substance Pinellia ternata lectin(PTL) protein in Pinelliae Rhizoma and the related processed products, this study prepared specific monoclonal antibodies against PTL by hybridoma cell technology, and established a quantitative double-antibody sandwich enzyme linked immunosorbent assay(ELISA) for PTL antigen. The detection conditions were 2.5 μg·mL~(-1) working concentration of the captured antibody and 1∶450 of the dilution multiple of detected antibody. The coating condition was staying overnight at 4 ℃. The blocking time and incubation times of antigen and detected antibody were all 90 minutes. The incubation time of horseradish peroxidase conjugated streptavidin-horseradish peroxidase(SA-HRP) was 15 minutes. The quantitative limit of the method for PTL antigen was 0.375 ng·mL~(-1). The linear range was 75.000-4 800.000 pg·mL~(-1), and R~2=0.997 1. The recovery rate was 90.0%-110.0%, and the variation coefficients of intra-test and inter-test precision were 2.0%-3.0% and 2.0%-8.5%.The content of PTL in three batches of Pinelliae Rhizoma and the related processed products was determined by the method, and the average content of PTL in Pinelliae Rhizoma was 35.42 mg·g~(-1). The average content of PTL in Pinelliae Rhizoma Praeparatum Cum Alumine, Pinelliae Rhizoma Praeparatum, and Pinelliae Rhizoma Praeparatum Cum Zingibere Et Alumine were 1.15 mg·g~(-1), 16.53 μg·g~(-1), and 122.63 ng·g~(-1), respectively, indicating that the content of PTL decreased significantly after processing. The quantitative double-antibody sandwich ELISA for PTL antigen established in this paper had good linearity, sensitive response, and high accuracy, which provided a simple and effective monitoring method for the detection of PTL content in the processing of Pinelliae Rhizoma.

Irritant toxicity and lectin content of different processed products of Pinelliae Rhizoma

The present study aims to investigate the correlation between irritant toxicity variation and lectin content variation during the processing of Pinelliae Rhizoma products and to explore the feasibility of Western blot as a method for the detection of lectin. We processed Pinelliae Rhizoma Praeparatum Cum Alumine, Pinelliae Rhizoma Praeparatum, and Pinelliae Rhizoma Praeparatumcum Zingibere et Alumine to different degrees and then analyzed their irritant toxicity via Draize rabbit eye test. Western blot was employed to determine the lectin content in Pinelliae Rhizoma products processed with different methods. The correlation between toxicity variation and lectin content variation was then analyzed. Different decoction pieces of Pinelliae Rhizoma were collected for the determination of lectin content. The three processed products of Pinelliae Rhizoma showed gradually decreased toxicity and lectin content as the processing continued. The decreasing trend of lectin content was consistent with that of irritant toxicity during processing, which indicated that the change in lectin content could reflect the trend of irritant toxicity. No band of lectin appeared in the Western blot of processed products of Pinelliae Rhizoma, which suggested that western blotting can be used for the detection of toxic lectin in the processed products of Pinelliae Rhizoma. Lectin should not be detected in the Pinelliae Rhizoma products processed according to the methods in the Chinese Pharmacopoeia.

Spectrum-effect relationship between HPLC fingerprints and inhibitory activity in MUC5AC mucin of Pinelliae Rhizoma Praeparatum Cum Alumine

Purpose: To investigate the spectrum-effect relationship between HPLC fingerprints and the inhibitory effect on MUC5AC mucin of Pinelliae Rhizoma Praeparatum Cum Alumine (PRPCA).Methods: The fingerprints of 20 PRPCA batches were established using HPLC and their similarities or differences were analyzed using hierarchical cluster analysis (HCA) and principal component analysis (PCA). The inhibitory effects of MUC5AC mucin were evaluated in LPS-treated NCI-H292 cells. The spectrum-effect relationship between common chromatographic peaks and MUC5AC inhibition was established using a partial least squares-discriminant analysis (PLS-DA).Results: Fifteen common chromatographic peaks were identified by analyzing HPLC fingerprints, with uridine, tyrosine, uracil, and inosine found as possible markers to distinguish the PRPCA from different sources. Spectrum-effect relationship analysis showed that the chromatographic peaks 5, 6, 10 (vernine), 12 (5-hydroxymethylfurfural), 14 (tryptophan) and 15 (adenosine) were closely associated with the inhibitory effect on MUC5AC mucin.Conclusion: The spectrum-effect relationship between HPLC fingerprints and the inhibitory effect on MUC5AC mucin of PRPCA was successfully established in the present study. Our findings further reveal the material basis of PRPCA and provide an effective method for its quality control.

Research Progress on Quality Control Methods for Xiaochaihu Preparations

Xiaochaihu (XCH) is a classic Chinese medicine formula. XCH tablet, XCH granule, XCH capsule, and XCH effervescent tablet are included in the Chinese Pharmacopoeia. In this review, the formula and quality standards of XCH preparations at home and abroad were compared. The differences in manufacturing process of XCH preparations are discussed. The progress of research on the qualitative identification, quantitative detection and fingerprint chromatogram/specific chromatogram of XCH preparations was reviewed. The characteristic components of Pinelliae Rhizoma Praeparatum Cum Zingibere Et Alumine and Jujubae Fructus was rarely analyzed for XCH preparations. It is suggested that the specificity of drug quality detection methods should be improved. Considering drug safety and drug efficacy, it is suggested to set the upper and lower limits of the content of saikosaponins. The standards for heavy metals and other limited items for XCH preparations are also suggested to be set.

Study on drug properties and syndrome-symptom-formula-herb network of traditional Chinese medicine in treatment of effort angina pectoris based on data visualization

This article aims to explore drug properties and syndrome-symptom-formula-herb network of traditional Chinese medicine(TCM) in the treatment of effort angina pectoris based on data visualization, and provide useful references for clinical diagnosis and treatment. Literatures about TCM formula for effort angina pectoris from databases of CNKI, WanFang, VIP, and CBM were retrieved from the database-building to August 31, 2019. The name of syndromes, symptoms, formulas, and herbs were standardized, and the corresponding databases were established. Frequency, four properties, five flavors, and meridian were analyzed. Visualized syndrome-symptom-formula-herb network relationships were constructed by bioinformatic analysis. A total of 202 formulas were included, and 218 kinds of TCM were involved. There were 56 herbs with the use frequency of more than 10, involving 78 syndromes and 162 symptoms. TCM formulas in the treatment of effort angina pectoris mainly included herbs with effects in invigorating blood circulation and eliminating stasis, tonifying deficiency, Qi-regulating, resolving phlegm and relieving cough and asthma, relieving exterior disorder, and heat-clearing. The main properties were warm, cold and mild(accounting for 95%); the main flavors were sweet, bitter and pungent(accounting for 89%); and meridians were mainly spleen, heart, liver, lung, stomach, and kidney(accounting for 89%). Syndrome-symptom-formula-herb network of TCM in the treatment of effort angina pectoris were successfully constructed. The high-frequency syndromes of this disease were Qi deficiency and blood stasis, Qi stagnation and blood stasis, heart blood stasis, and turbid phlegm and blood stasis, and its high-frequency symptoms were chest tightness, chest pain, palpitation, shortness of breath, fatigue, dark purple tongue, spontaneous sweating, and abundant phlegm. High-frequency core formulas of this disease included Xuefu Zhuyu Decoction, Gualou Xiebai Banxia Decoction, Danshen Decoction, Taohong Siwu Decoction, Shengmai Powder, Buyang Huanwu Decoction and Zhigancao Decoction, and their core herbs included Salviae Miltiorrhizae Radix et Rhizoma, Astragali Radix, Chuanxiong Rhizoma, Ginseng Radix et Rhizoma, Trichosanthis Fructus, Allium Macrostemonis Bulbus, Notoginseng Radix et Rhizoma, Persicae Semen, Carthami Flos, Atractylodis Macrocephalae Rhizoma, Angelicae Sinensis Radix, Paeoniae Radix Rubra, Poria, Pinelliae Rhizoma Praeparatum. Drug properties and syndrome-symptom-formula-herb networks of TCM in the treatment of effort angina pectoris can realize data visualization, objectively reflect the clinical syndrome differentiation and rule of medication, and provide reference for clinical diagnosis and treatment.

Effect of Pinelliae Rhizoma processed with lime and licorice on toxic lectin protein

The present study was aimed to investigate the effect of lime and licorice processing of Pinelliae Rhizoma on its toxic lectin protein and clarify the scientific detoxification connotation of lime and licorice processing of Pinelliae Rhizoma. Western blot was used to semi-quantitatively analyze the contents of lectin in Pinelliae Rhizoma and Pinelliae Rhizoma Praeparatum. Raw products and lectin were treated by soaking in licorice juice, lime solution or mixture solution of these two to investigate the different processing time on the content of toxic lectin protein. SDS-PAGE gel electrophoresis was used to analyze the changes of lectin protein bands in the solution and precipitates before and after processing. MALDI-TOF technology was used to qualitatively analyze and compare the protein molecular weight before and after processing. The results showed that the contents of lectin in Pinelliae Rhizoma and Pinelliae Rhizoma Praeparatum were 5.01% and 0.04% respectively, indicating that processing could significantly reduce the content of active lectin in raw products. The results also showed that the content of lectin in raw drugs decreased significantly after soaking in lime solution for one day or in licorice juice for three day, and the effect was greatest in mixture solution. Qualitative analysis showed that after being treated by soaking in lime solution, the lectin protein was decomposed into small peptide segments, while after being treated by soaking in licorice juice, the lectin protein was denatured and precipitated. The structure of lectin protein in Pinelliae Rhizoma was broken after being processed with licorice juice and lime solution, which significantly reduced the content of toxic lectinprotein. This is one of the detoxification mechanisms of Pinelliae Rhizoma processing.

Effect of processing on toxic components lectin from four kinds of Araceae toxic medicines

The study aimed to investigate the effect of processing on lectin protein in four toxic Chinese medicines tubers of Pinellia ternata,P. pedatisecta,Arisema heterophyllum and Typhonium giganteum. Western blot was used to semi-quantitatively analyze the content of lectin in the four kinds of toxic Chinese medicines and their different processed products. Raw products and lectin were treated by heating or soaking in ginger juice or alum solution. The effects of different excipients and the heating methods on lectin proteins were investigated. The results showed that the content of lectin in raw products of P. pedatisecta,P. ternata,A. heterophyllum,and T. giganteum were 7. 3%,4. 9%,2. 7%,2. 3%,respectively. And the content of lectin in Pinelliae Rhizoma praeparatum cum alumine was 0. 027%. Lectin was not detected in the Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine,Arisaematis Rhizma Praeparatum and Typhonii Rhizoma Praeparatum,which indicated that processing could significantly reduce the content of active lectin in raw products. The results also showed that with the prolongation of soaking and heating time,the content of lectin in raw products decreased gradually,while the content was almost unchanged when soaked in ginger juice alone. The effects of different excipients and heating on lectin were the same as those on raw products. Therefore,the method with alum soaking and heating can reduce the content of active lectin,which is the key to reduce the toxicity of toxic Chinese medicines. In this paper,Western blot was used to study the content of toxic protein in Araceae toxic Chinese medicines as an evaluation method of the processing degree.

Purification, Characterization of Two Polysaccharides from Pinelliae Rhizoma Praeparatum Cum Alumine and Their Anti-Inflammatory Effects on Mucus Secretion of Airway Epithelium.

Pinelliae Rhizoma Praeparatum cum Alumine (PRPCA) is an important traditional processed herbal medicine mainly used for treating phlegm in China for more than 2000 years. In our previous studies, extraction optimization, characterization, and bioactivities of total polysaccharides from PRPCA were investigated. In this study, further purification of these polysaccharides was performed. Two polysaccharides named neutral fraction of total polysaccharides-II (TPN-II) and acidic fraction of total polysaccharides-II (TPA-II) were obtained by gradient ion-exchange chromatography followed by gel-permeation chromatography. Results of scanning electron microscopy (SEM) analysis in the present study showed that TPN-II had a tight structure with a rough and uneven surface, while TPA-II had a relative homogeneous surface and a loose structure. Further studies indicated that TPN-II was a homosaccharide mainly composed by glucose with a molecular weight of 8.0 kDa. TPA-II was mainly composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose and arabinose in a molar ratio of 2.1, 2.3, 1.7, 10.6, 2.6, 14.2, and 2.5, with a molecular weight of 1250 kDa. The nuclear magnetic resonance (NMR) results indicated that α and β form glycoside bonds existed in TPN-II and TPA-II, and TPN-II was composed of α-glucopyranose. In addition, both purified polysaccharides have significant anti-inflammatory effects on mucus secretion of human airway epithelial NCI-H292 cells without cytotoxicity. Compared with TPN-II, TPA-II exhibited more significant anti-inflammatory effects on lipopolysaccharide (LPS)-induced airway inflammation by regulating levels of interleukin-4 (IL-4) and interferon-γ (IFN-γ) and inhibiting mucus secretion. The results suggest that polysaccharides from PRPCA could be explored as therapeutic agents in treating inflammation and over secretion of mucus in asthma.

Pinelliae Rhizoma Praeparatum Cum Alumine Extract: Sedative and Hypnotic Effects in Mice and Component Compounds.

Pinelliae Rhizoma Praeparatum Cum Alumine (PRPCA) is useful for eliminating dampness and phlegm in clinical settings, targeting the main mechanisms of insomnia as defined in traditional Chinese medicine. However, little is known regarding the sedative and hypnotic effects of PRPCA. In the present study, we examined the sedative effects of PRPCA via a locomotor activity test and aimed to determine the most appropriate concentration of PRPCA for achieving these effects. The strongest sedative effects were observed at a PRPCA concentration of 0.45 g/ml. In addition, we investigated the hypnotic effects of PRPCA and its role in promoting sleep via sleep monitoring and vigilance state analysis. PRPCA increased rapid eye movement (REM) sleep and non-REM (NREM) sleep while decreasing wakefulness. In addition, PRPCA decreased the number of bouts of wakefulness (16–32 s and 32–64 s) and increased the number of bouts of NREM sleep (128–256 s). Furthermore, we identified a total of 32 component compounds via chromatography and mass spectrometry. Hence, the current work provides valuable information regarding the sedative and hypnotic effects of PRPCA and its regulatory mechanisms in promoting sleep.

Comparative research of the curative effects of pinelliae rhizoma and pinelliae rhizoma praeparatum cum alumine on ovalbumin-induced allergic asthma in rats

Aim: This study was designed to compare the curative effects of Pinelliae Rhizoma (RPT) and Pinelliae Rhizoma Praeparatum cum Alumine (QPT) on ovalbumin (OVA)-induced allergic asthma in rats. Materials and Methods: Asthmatic rats were prepared by intraperitoneal injection and inhalation of OVA, and RPT and QPT at the doses of 120 and 240 mg/kg/day were administered orally for consecutive 7 days. Then, histopathological changes of the lung tissues were examined; levels of interleukin (IL)-4 and interferon (IFN)-γ in the bronchoalveolar lavage fluids (BALFs) and immunoglobulin (Ig) E in the serum were measured by enzyme-linked immunosorbent assay; eosinophils (EOS) in BALFs were also determined. Furthermore, the expressions of mucin 5AC (MUC5AC) and AQP-5 in the lung tissues were determined by quantitative real-time fluorogenic polymerase chain reaction and western blot assays. Besides, high-performance liquid chromatography-LTQ-Orbitrap-Elite-MS analysis was applied to compare the constituent's difference between the RPT and QPT. Results: The present results revealed that both the RPT and QPT have the potential curative effects for treating OVA-induced asthma in rats via inhibiting inflammatory reactions (suppressing EOS influx, IgE secretion, and release of IL-4 whereas increasing the release of IFN-γ) and modulating the mRNA and protein expressions of MUC5AC and AQP-5. In addition, our results also evidenced that QPT exhibited a better curative effect on allergic asthmatic rats compared to RPT. Conclusion: Our results are beneficial for confirming the traditional Chinese medicinal theory of “QPT is good at removing dampness to reduce phlegm.” Abbreviations used: BALF: Bronchoalveolar lavage fluid; ELISA: Enzyme-linked immunosorbent assay; EOS: Eosinophils; IFN: Interferon; IL: Interleukin; OVA: Ovalbumin; qRT-PCR: Quantitative real-time fluorogenic polymerase chain reaction; QPT: Rhizoma Pinelliae Praeparatum cum Alumine; RIPA: Radioimmunoprecipitation assay; RPT: Pinelliae Rhizoma; SDS-PAGE: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis; TCM: Traditional Chinese medicine.

Pinelliae Rhizoma Praeparatum Involved in the Regulation of Bile Acids Metabolism in Hepatic Injury

Pinelliae Rhizoma Praeparatum (PRP) as traditional Chinese medicine had been used for hepatic diseases in combinative forms. However, the effect of PRP was not clear when used alone. So to explore the hepatoprotective/hepatotoxin of PRP is necessary. The activities of PRP were investigated in acetaminophen-induced hepatic injury mice. Liver function markers, hepatic oxidative stress markers were evaluated. Bile acids metabolic transports and nuclear factor erythroid 2-related factor 2 (Nrf2) were detected. As a drug for the treatment of liver diseases, PRP slightly restored the parameters towards normal in model mice only in low dosage, and also had no antioxidant activity and regulate Nrf2. Cholestasis was significantly elevated in model mice when pretreatment with routine or high dosage of PRP, but had no effect on normal mice. Bile salt export pump (Bsep) and multidrug resistance-associated protein 2 (Mrp2) in model mice were markedly increased when pretreatment with low dose PRP, but significantly decreased when pretreatment in routine or high dosage. Mrp3 was significantly induced in model mice after pretreatment of PRP. But the adjustment effect to bile acids transporters by PRP was not significant in normal mice. These results reveal that PRP has the different effects on bile acids transporter in hepatic injury mice, and therefore, the dosage of PRP need to be paid attention to when it is used in clinical hepatic injury.

Pharmacokinetics and pharmacodynamics of citrus peel extract in lipopolysaccharide-induced acute lung injury combined with Pinelliae Rhizoma Praeparatum.

Dry citrus peel (Chenpi) is not only consumed as a dietary supplement, but also used for the treatment of respiratory diseases. Pinelliae Rhizoma Praeparatum (Banxia) is always used with Chenpi for the treatment of respiratory diseases, but β-sitosterol, the main active component in Banxia, as a food additive, shows no respiratory system activity. In the present study, the pharmacokinetic characters showed that the absorption of the active components of Chenpi was accelerated under pathological conditions combined with Banxia. Although the bioavailability of active components did not significantly change, the distribution of active components in tissues increased, particularly in the target organ. These results are consistent with the combination of Chenpi with β-sitosterol. Furthermore, the pharmacodynamics result also indicated that Chenpi combined with Banxia or β-sitosterol was able to ameliorate histopathologic damage and decrease the levels of inflammatory factors. The results suggest that pharmacokinetic interactions improve the pharmacological activity of Chenpi in respiratory diseases.

Extraction Optimization, Characterization, and Bioactivities of Polysaccharides from Pinelliae Rhizoma Praeparatum Cum Alumine Employing Ultrasound-Assisted Extraction.

In this study, the ultrasound-assisted extraction of polysaccharides (PSA) from Pinelliae Rhizoma Praeparatum Cum Alumine (PRPCA) was optimized by response surface methodology (RSM). The structural characteristics of PSA were analyzed by UV-vis spectroscopy, infrared spectroscopy, scanning electron microscopy, high performance gel permeation chromatography and high performance liquid chromatography, respectively. In addition, antioxidant and antimicrobial activities of PSA were studied by different in vitro assays. Results indicated that the optimal extraction conditions were as follows: the ratio of water to raw of 30 mL/g, extraction time of 46.50 min, ultrasonic temperature of 72.00 °C, and ultrasonic power of 230 W. Under these conditions, the obtained PSA yield (13.21 ± 0.37%) was closely agreed with the predicted yield by the model. The average molecular weights of the PSA were estimated to be 5.34 × 103 and 6.27 × 105 Da. Monosaccharide composition analysis indicated that PSA consisted of mannose, galactose uronic acid, glucose, galactose, arabinose with a molar ratio of 1.83:0.55:75.75:1.94:0.45. Furthermore, PSA exhibited moderate antioxidant and antibacterial activities in vitro. Collectively, this study provides a promising strategy to obtain bioactive polysaccharides from processed products of herbal medicines.

Standardization of the manufacturing procedure for Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine

Ethnopharmacological relevancePinelliae Rhizoma (PR), the dried tuber of Pinellia ternata (Thunb.) Breit., is a traditional Chinese medicinal herb. It is commonly used for treating cancer, cough and phlegm. To treat cancer, Chinese medicine practitioners often use raw PR; while to treat cough and phlegm, they usually use Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine (PRZA, raw PR processed with ginger juice and alumen as adjuvant materials). Currently, the producing protocol of PRZA varies greatly among different places in China. This study aims to standardize the manufacturing procedure for PRZA. We also evaluated the impact of processing on the bioactivities and chemical profile of raw PR. Materials and methodsWe used the orthogonal design to optimize the manufacturing procedure of PRZA at bench scale, and validated the optimized procedure in pilot-scale production. The MTT assay was used to compare the cytotoxicities of raw PR and PRZA in hepatocellular carcinoma HepG2 cells. Animal models (ammonia liquor-induced cough model and phenol red secretion model) were used to compare the antitussive and expectorant effects of raw PR and PRZA, respectively. The chemical profiles of raw PR and PRZA samples were compared using a newly developed ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS) method. ResultsThe standardized manufacturing procedure for PRZA is as follows: soak raw PR in water until the center of the cut surface is devoid of a dry core, after that, boil the herb in water (for each 100kg raw PR, 12.5kg alumen and 25L freshly squeezed ginger juice are added) for 6h, and then take out and dry them. The cytotoxicity of PRZA was less potent than that of raw PR. Intragastric administration of raw PR or PRZA demonstrated antitussive and expectorant effects in mice. These effects of PRZA were more potent than that of raw PR at the dose of 3g/kg. By comparing the chemical profiles, we found that six peaks were lower, while nine other peaks were higher in PRZA than in raw PR. Six compounds corresponding to six individual changed peaks were tentatively identified by matching with empirical molecular formulae and mass fragments. ConclusionThe manufacturing procedure for PRZA was standardized. This protocol can be used for PRZA industrial production. The bioactivity assay results of raw PR and PRZA (produced using the standardized protocol) support the common practice for the clinical applications of these two decoction pieces. Moreover, raw PR and PRZA showed different chemical profiles. Further studies are warranted to establish the relationship between the alteration of chemical profiles and the changes of medicinal properties caused by processing.

Pinelliae rhizoma, a toxic chinese herb, can significantly inhibit CYP3A activity in rats.

Raw Pinelliae Rhizoma (RPR) is a representative toxic herb that is widely used for eliminating phlegm or treating cough and vomiting. Given its irritant toxicity, its processed products, including Pinelliae Rhizoma Praeparatum (PRP) and Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine (PRPZA), are more commonly applied and administered concomitantly with other chemical drugs, such as cough medications. This study aimed to investigate the effects of RPR, PRP, and PRPZA on CYP3A activity. Testosterone (Tes) and buspirone (BP) were used as specific probe substrates ex vivo and in vivo, respectively. CYP3A activity was determined by the metabolite formation ratios from the substrates. Ex vivo results show that the metabolite formation ratios from Tes significantly decreased, indicating that RPR, PRP, and PRPZA could inhibit CYP3A activity in rats. CYP3A protein and mRNA levels were determined to explore the underlying mechanism. These levels showed marked and consistent down-regulation with CYP3A activity. A significant decrease in metabolite formation ratios from BP was also found in PRPZA group in vivo, implying that PRPZA could inhibit CYP3A activity. Conclusively, co-administration of PR with other CYP3A-metabolizing drugs may cause drug–drug interactions. Clinical use of PR-related formulae should be monitored carefully to avoid adverse interactions.

Comparison of Chemical Composition in Decoctions of Aconitum kusnezoffii Reichb and Pinelliae rhizoma Praeparatum Extracted by Different Methods

Comparison of Chemical Composition in Decoctions of Aconitum kusnezoffii Reichb and Pinelliae rhizoma Praeparatum Extracted by Different Methods