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Background: Different parts of radish, such as the roots, leaves, and seeds, can be used in culinary and traditional medicine treatments in East Asia. Objectives: The aim of this study was to explore the difference of Raphani Semen (RS) which is sourced from radishes (R) with red skin (RRS), green skin (GRS), and white skin (WRS) through their appearance and chemical characteristics. Materials and Methods: A total of 25 batches of RRS, GRS, and WRS products were collected. The length, width, thickness, shape, flavor, and skin color of RS were carried out using vernier caliper and flavor rating assessment, respectively. The HPLC-DAD method was used to establish fingerprint analysis and measure the contents of sinapine thiocyanate and 3,6'-disinapoyl sucrose. Results: It was shown that the appearance and content varied in RRS, GRS, and WRS. In addition, the method of PLS-DA and LDA can differentiate RRS, GRS, and WRS. Conclusion: RS is diverse, and the appearance and chemical characteristics can be used to differentiate RRS, WRS, and GRS. [ABSTRACT FROM AUTHOR]

Background. Intervertebral disc degeneration (IDD) is a major cause of low back pain. Sinapine thiocyanate (ST) has been reported to have a wide range of biological activities. However, the treatment of IDD with ST has not been studied. Objectives. To explore the role and mechanism of ST treatment in IDD. Materials and methods. Nucleus pulposus cells (NPCs) were induced using lipopolysaccharide (LPS), which was used as an in vitro model of IDD. Cell activity, oxidative stress-related indicators and protein expression were detected using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, enzyme-linked immunosorbent assay (ELISA) and western blot. Pyroptosis was evaluated with propidium iodide (PI)/Hoechst double staining and immunofluorescence for NOD-like receptor protein 3 (NLRP3), and pyroptosis-related proteins and inflammatory factors were measured with western blot and ELISA. The pathological changes of IDD were assessed with hematoxylin & eosin (H&E) and safranin-O staining. Results. Our results showed that ST alleviated LPS-induced degeneration of NPCs, as evidenced by reducing reactive oxygen species (ROS), malondialdehyde (MDA), matrix metalloproteinase-13 (MMP-13), a disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5), and increasing collagen II and aggrecan expression. Moreover, ST repressed LPS-induced pyroptosis by inhibiting NLRP3, caspase-1 p20, interleukin (IL)-1β and IL-18. Further studies showed that ST did not restrain the activation of the JAK1/STAT3 signaling pathway induced by colivelin, or of the enhanced pyroptosis induced by polyphyllin VI. Sinapine thiocyanate alleviated IDD in vivo and suppressed NLRP3-mediated pyroptosis and the JAK1/STAT3 signaling pathway. Conclusions. Sinapine thiocyanate could alleviate IDD, although this did not include a reduction in NLRP3- mediated pyroptosis and inactivation of the JAK1/STAT3 signaling pathway, thus potentially being a candidate drug for IDD treatment. [ABSTRACT FROM AUTHOR]

Sinapine thiocyanate (ST), an alkaloid existed extensively in seeds of cruciferous plants, exhibits a number of pharmacological effects, including anti-inflammatory and anti-malignancy properties. However, it is still unknown what effects and molecular mechanisms ST has on colorectal cancer (CRC). In the current study, it was indicated that ST inhibited proliferation, colony formation, and apoptosis in vitro, as well as arrested the G1 phase of CRC cells. There was a significant repressive effects of ST on invasion and migration of CRC cells in vitro. RNA-sequencing indicated that 750 differentially expressed genes existed in CRC cells after ST treatment, and enrichment analysis demonstrated that ST obviously decreased the activation of keratinization pathways. Among DEGs enriched in keratinization, keratin 6A (KRT6A) was decreased the most significant, as well as its target gene S100 calcium-binding protein A2 (S100A2). Low expression of KRT6A and S100A2 signatures indicated a favorable prognosis in CRC patients. Moreover, we found overexpression of KRT6A relieved the inhibitory effects of ST in CRC cells. Furthermore, ST inhibited the CRC cell proliferation in vivo, and reduced KRT6A and KI67 expression in xenograft tumor. Taken together, we demonstrated that ST exhibited anti-CRC properties by inhibiting KRT6A/S100A2 axis. It is possible that ST can be used as a treatment for CRC.

Sinapine thiocyanate (ST), an alkaloid existed extensively in seeds of cruciferous plants, exhibits a number of pharmacological effects, including anti-inflammatory and anti-malignancy properties. However, it is still unknown what effects and molecular mechanisms ST has on colorectal cancer (CRC). In the current study, it was indicated that ST inhibited proliferation, colony formation, and apoptosis in vitro, as well as arrested the G1 phase of CRC cells. There was a significant repressive effects of ST on invasion and migration of CRC cells in vitro. RNA-sequencing indicated that 750 differentially expressed genes existed in CRC cells after ST treatment, and enrichment analysis demonstrated that ST obviously decreased the activation of keratinization pathways. Among DEGs enriched in keratinization, keratin 6A (KRT6A) was decreased the most significant, as well as its target gene S100 calcium-binding protein A2 (S100A2). Low expression of KRT6A and S100A2 signatures indicated a favorable prognosis in CRC patients. Moreover, we found overexpression of KRT6A relieved the inhibitory effects of ST in CRC cells. Furthermore, ST inhibited the CRC cell proliferation in vivo, and reduced KRT6A and KI67 expression in xenograft tumor. Taken together, we demonstrated that ST exhibited anti-CRC properties by inhibiting KRT6A/S100A2 axis. It is possible that ST can be used as a treatment for CRC. [ABSTRACT FROM AUTHOR]

Background: Intervertebral disc degeneration (IDD) is a major cause of low back pain. Sinapine thiocyanate (ST) has been reported to have a wide range of biological activities. However, the treatment of IDD with ST has not been studied., Objectives: To explore the role and mechanism of ST treatment in IDD., Material and Methods: Nucleus pulposus cells (NPCs) were induced using lipopolysaccharide (LPS), which was used as an in vitro model of IDD. Cell activity, oxidative stress-related indicators and protein expression were detected using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, enzyme-linked immunosorbent assay (ELISA) and western blot. Pyroptosis was evaluated with propidium iodide (PI)/Hoechst double staining and immunofluorescence for NOD-like receptor protein 3 (NLRP3), and pyroptosis-related proteins and inflammatory factors were measured with western blot and ELISA. The pathological changes of IDD were assessed with hematoxylin & eosin (H&E) and safranin-O staining., Results: Our results showed that ST alleviated LPS-induced degeneration of NPCs, as evidenced by reducing reactive oxygen species (ROS), malondialdehyde (MDA), matrix metalloproteinase-13 (MMP-13), a disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5), and increasing collagen II and aggrecan expression. Moreover, ST repressed LPS-induced pyroptosis by inhibiting NLRP3, caspase-1 p20, interleukin (IL)-1β and IL-18. Further studies showed that ST did not restrain the activation of the JAK1/STAT3 signaling pathway induced by colivelin, or of the enhanced pyroptosis induced by polyphyllin VI. Sinapine thiocyanate alleviated IDD in vivo and suppressed NLRP3-mediated pyroptosis and the JAK1/STAT3 signaling pathway., Conclusions: Sinapine thiocyanate could alleviate IDD, although this did not include a reduction in NLRP3-mediated pyroptosis and inactivation of the JAK1/STAT3 signaling pathway, thus potentially being a candidate drug for IDD treatment.

[Display omitted] • A green and earth-friendly method for the extraction of sinapine thiocyanate from black mustard seeds was developed. • The synergy between ultrasound and natural deep eutectic solvent in the extraction process was clarified. • Highest extraction rate with mixtures of choline chloride/ethylene glycol (20% water). Herein, several choline chloride-based (ChCl) natural deep eutectic solvents (NADESs) were investigated for the ultrasonic-assisted extraction (UAE) of sinapine thiocyanate (ST) from black mustard seed (BMS). The NADES consisting of choline chloride and ethylene glycol (EG) was screened as the extraction solvent and single-factor experiments were carried out. In addition, compared with the conventional extraction solvents and extraction methods. The optimized extraction conditions were described below: NADES synthesized by ChCl and EG was used as an extractant with a water content of 20 %, and the solid–liquid ratio was 1:30 g∙mL−1. The mixture was extracted under the ultrasonic parameters of 60 ℃ and 200 W for 75 min. The extraction rate obtained using this method was higher than traditional solvents such as methanol, ethanol and water, and the time consumed was also greatly reduced compared to heated reflux extraction. A variety of instruments were used to characterize the experimental materials and further elucidate the extraction mechanism. Meanwhile, the antioxidant capacity of the ChCl/EG extract of BMS was determined by ultraviolet–visible spectrophotometry. The green evaluation of the method was carried out by the complex green analytical procedure index (ComplexGAPI). This work provides a green technique for valid extraction of ST from BMS, which is expected to be expanded to pharmaceutical, food and other related industries. [ABSTRACT FROM AUTHOR]

The increase of blood pressure is accompanied by the changes in the morphology and function of vascular endothelial cells. Vascular endothelial injury and hypertension actually interact as both cause and effect. A large number of studies have proved that inflammation plays a significant role in the occurrence and development of hypertension, but the potential mechanism between inflammation and hypertensive endothelial injury is still ambiguous. The purpose of this study was to explore the association between the activation of NLRP3 inflammasome and hypertensive endothelial damage, and to demonstrate the protective effect of sinapine thiocyanate (ST) on endothelia in hypertension. The expression of NLRP3 gene was silenced by tail vein injection of adeno-associated virus (AAVs) in spontaneously hypertensive rats (SHRs), indicating that activation of NLRP3 inflammasome accelerated hypertensive endothelial injury. ST not only protected vascular endothelial function in SHRs by inhibiting the activation of NLRP3 inflammasome and the expression of related inflammatory mediators, but also improved AngII-induced huvec injury. In summary, our results show that alleviative NLRP3 inflammasome activation attenuates hypertensive endothelial damage and ST ameliorates vascular endothelial dysfunction in hypertension via inhibiting activation of the NLRP3 inflammasome.

Sinapine thiocyanate (ST), an alkaloid existed extensively in seeds of cruciferous plants, exhibits a number of pharmacological effects, including anti-inflammatory and anti-malignancy properties. However, it is still unknown what effects and molecular mechanisms ST has on colorectal cancer (CRC). In the current study, it was indicated that ST inhibited proliferation, colony formation, and apoptosis in vitro , as well as arrested the G1 phase of CRC cells. There was a significant repressive effects of ST on invasion and migration of CRC cells in vitro . RNA-sequencing indicated that 750 differentially expressed genes existed in CRC cells after ST treatment, and enrichment analysis demonstrated that ST obviously decreased the activation of keratinization pathways. Among DEGs enriched in keratinization, keratin 6A (KRT6A) was decreased the most significant, as well as its target gene S100 calcium-binding protein A2 (S100A2). Low expression of KRT6A and S100A2 signatures indicated a favorable prognosis in CRC patients. Moreover, we found overexpression of KRT6A relieved the inhibitory effects of ST in CRC cells. Furthermore, ST inhibited the CRC cell proliferation in vivo , and reduced KRT6A and KI67 expression in xenograft tumor. Taken together, we demonstrated that ST exhibited anti-CRC properties by inhibiting KRT6A/S100A2 axis. It is possible that ST can be used as a treatment for CRC.

The hepatoprotective effect of Brassica rapa seeds was evaluated. The antioxidant activities of extracts were determined using total phenolic content, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, oxygen radical absorbance capacity (ORAC) and reducing power assays. Hepatoprotective effects were evaluated using CCl4-induced mouse liver damage animal model. Identification and quantification of the compound were based on ultra performance liquid chromatographic (UPLC) analysis. All extracts of Brassica rapa seeds exhibited high phenolic contents and strong antioxidant activities. The activities were in the order of ethanol extract ≥ 50% ethanol extract>water extract. All the three extracts showed effective hepatoprotection. They could ameliorate liver damage and improve the antioxidant defense system. Sinapine thiocyanate was the main component and was hepatoprotective in a dose-dependent manner. These findings imply that Brassica rapa seeds could be developed as a functional food for hepatoprotection.

The aim of the present study was to investigate the effect and the underlying mechanism of the combined treatment of rhynchophylla total alkaloids (RTA) and sinapine thiocyanate for protection against a prothrombotic state (PTS) associated with the tumor necrosis factor-alpha (TNF-a)-induced inflammatory injury of vascular endothelial cells (VECs). A TNF-a-induced VEC inflammatory injury model was established, and cell morphology of VECs was evaluated using scanning electron microscopy. In addition, reverse transcription-quantitative polymerase chain reaction and western blot analysis were performed to examine the mRNA and protein expression of coagulation-related factors, including nuclear factor-κB (NF-κB), transforming growth factor-ß1 (TGF-ß1), tissue factor (TF), plasminogen activator inhibitor (PAI-1), protease-activation receptors (PAR-1) and protein kinase C (PKC-a) in VECs. Combined treatment with RTA and sinapine thiocyanate was demonstrated to reduce, to a varying extent, the mRNA and protein expression of NF-κB, TGF-ß1, TF, PAR-1, PKC-a and PAI-1. Furthermore, combined treatment with RTA and sinapine thiocyanate was able to downregulate the expression of coagulation-related factors in injured VECs, thereby inhibiting the PTS induced by vascular endothelial injury. The underlying mechanism is partially associated with the TF-mediated activation of the thrombin-receptor signaling pathway that suppresses coagulation during inflammation and balances fibrinolysis in order to inhibit fibrin generation and deposition. [ABSTRACT FROM AUTHOR]

The hepatoprotective effect of Brassica rapa seeds was evaluated. The antioxidant activities of extracts were determined using total phenolic content, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, oxygen radical absorbance capacity (ORAC) and reducing power assays. Hepatoprotective effects were evaluated using CCl 4 -induced mouse liver damage animal model. Identification and quantification of the compound were based on ultra performance liquid chromatographic (UPLC) analysis. All extracts of Brassica rapa seeds exhibited high phenolic contents and strong antioxidant activities. The activities were in the order of ethanol extract ≥ 50% ethanol extract>water extract. All the three extracts showed effective hepatoprotection. They could ameliorate liver damage and improve the antioxidant defense system. Sinapine thiocyanate was the main component and was hepatoprotective in a dose-dependent manner. These findings imply that Brassica rapa seeds could be developed as a functional food for hepatoprotection. [ABSTRACT FROM AUTHOR]

A rapid and simple high-performance liquid chromatography ( HPLC)-photodiode array ( PDA) analytical method was developed for the quantitative analysis of Raphani Semen (RS). This method was successfully used to determine the five main phenolic compounds found in RS specimens from different production regions. The compounds included sinapine thiocyanate ( 1), β- d-fructofuranosyl-α- d-(6- O-sinapoyl)-glucopyranoside ( 2), isorhamnetin 3,4′-di- O-β- d-glucoside ( 3), β- d-(3- O-sinapoyl)-fructofuranosyl-α- d-(6- O-sinapoyl)-glucopyranoside ( 4), and β- d-(3,4- O-disinapoyl)-fructofuranosyl-α- d-(6- O-sinapoyl)-glucopyranoside ( 5). The marker compounds were separated using an Agilent Eclipse XDB-C18 column (5.0 µm, 150 × 4.6 mm i.d.) by gradient elution with acetonitrile/water/0.1% trifluoroacetic acid ( TFA) as the mobile phase (flow rate, 1.0 mL/min). This method was fully validated with respect to linearity, precision, accuracy, stability, and robustness. The HPLC analytical method was validated to conduct a pattern recognition analysis by repeatedly analyzing 56 seed samples including 55 RS ( C01-C49 and K50-K55) and 1 Brassicae Semen samples. In addition, a content standard for RS was proposed. Compounds 1 and 4 were revealed as major components in the HPLC chromatogram, and their contents ranged from 0.06 to 0.20 and 0.02 to 0.35 mg/g, respectively. These results demonstrate the successful development of an analytical method suitable for evaluating the quality and distinguishing the origin of RS. In addition, we briefly describe the crucial liquid chromatography-tandem mass spectrometry ( LC-MS/ MS) analytical conditions for the precise simultaneous quantification of the marker compounds. [ABSTRACT FROM AUTHOR]

Abstract: White mustard (Sinapis alba L.), a traditional Chinese medicine, is widely used in China for clinical prevention and treatment of the common winter diseases of asthma and bronchitis by percutaneous administration in the summer. The present study is to investigate the skin penetration behavior of white mustard extract to elucidate the possible mechanism underlying its immune regulation activity. The principle active compound of the extract, sinapine thiocyanate (ST), was used as a marker. The skin penetration of ST in white mustard extract was examined in vitro and in vivo. In vitro study on excised guinea pig hairless skin using Franz diffusion cell revealed ST can permeate through the skin and also accumulate in the skin. In vivo study was carried out on the guinea pig hairless skin for 24h, and then skin was excised for frozen section, ST from the sections were extracted to quantify the amount of drug in different skin layers. The detailed distribution of ST showed that it accumulated in the epidermis, especially in the stratum corneum. After treatment with white mustard extract for 24h, the skin was stained with ATPase, and the morphometric parameters of epidermal LCs were compared to the untreated control through image-analysis system. A statistically significant reduction in LC density and increase in shape factor were observed. Cytokines related to LCs migration including interleukin 1? (IL-1?) and tumor necrosis factor ? (TNF-?) were also measured after white mustard extract treated at different time points. Compared to the untreated group, white mustard extract significantly enhanced the release of IL-1? and TNF?. The morphometric changes of LCs and the local cytokine release after topical white mustard treatment may explain the activity of the white mustard extract against asthma and bronchitis. [Copyright &y& Elsevier]

• Metabolites of sinapine thiocyanate in plasma, urine and feces of rats were identified. • Metabolic profile includes extensive metabolisms of phase Ⅰ and phase Ⅱ. • A UHPLC-MS/MS method was developed to quantify sinapine thiocyanate in rat plasma. • The oral absolute bioavailability of sinapine thiocyanate was 1.84%. [Display omitted] Sinapine thiocyanate (ST) is an index component and pharmacological active component of Semen Sinapis and Semen Raphani , and it is widely used to relieving cough and asthma. This study aimed to obtain the metabolic and pharmacokinetic characterization of ST. The metabolic profiles of ST were obtained from rat plasma, urine, and feces via ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UHPLC-Q/TOF-MS). Thirteen metabolites were structurally identified, and the proposed metabolic pathways of ST included deamination, demethylation, hydrogenation, dehydration, and extensive conjugation, including glucuronidation and sulfonation. ST was selected as the plasma marker for the pharmacokinetic study. A simple and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for the quantitation of ST in rat plasma. The linear range of ST was 0.1–500 ng/mL (R 2 = 0.9976), and the lowest limit of quantification was 0.1 ng/mL. The intra-precision and inter-precision of the assay were 1.31–5.12% and 2.72–7.66%, and the accuracy (RE%) ranged from − 4.88% to 3.82% and − 3.47% to 6.18%. The extraction recovery, matrix effect, and stability of ST were within acceptable limits. The established method was validated and successfully applied to the pharmacokinetic study of ST. For pharmacokinetic experiments, the male Sprague-Dawley rats were administrated with ST solution intravenously (2 mg/kg) or orally (100 mg/kg). The oral absolute bioavailability of ST was calculated as 1.84%, and the apparent volume of distribution of intravenous and intragastric administrations were 107.51 ± 21.16 L/kg and 78.60 ± 14.44 L/kg, respectively. The maximum plasma concentration was 47.82 ± 18.77 nM, and the time to maximum peak was 88.74 ± 20.08 min for the intragastric dosing group. According to the pharmacokinetic and metabolic profiling results, metabolites with high abundance of ST in bio-fluids would be the next object in tissue distribution and pharmacodynamic study. [ABSTRACT FROM AUTHOR]

The increase of blood pressure is accompanied by the changes in the morphology and function of vascular endothelial cells. Vascular endothelial injury and hypertension actually interact as both cause and effect. A large number of studies have proved that inflammation plays a significant role in the occurrence and development of hypertension, but the potential mechanism between inflammation and hypertensive endothelial injury is still ambiguous. The purpose of this study was to explore the association between the activation of NLRP3 inflammasome and hypertensive endothelial damage, and to demonstrate the protective effect of sinapine thiocyanate (ST) on endothelia in hypertension. The expression of NLRP3 gene was silenced by tail vein injection of adeno-associated virus (AAVs) in spontaneously hypertensive rats (SHRs), indicating that activation of NLRP3 inflammasome accelerated hypertensive endothelial injury. ST not only protected vascular endothelial function in SHRs by inhibiting the activation of NLRP3 inflammasome and the expression of related inflammatory mediators, but also improved AngII-induced huvec injury. In summary, our results show that alleviative NLRP3 inflammasome activation attenuates hypertensive endothelial damage and ST ameliorates vascular endothelial dysfunction in hypertension via inhibiting activation of the NLRP3 inflammasome. [ABSTRACT FROM AUTHOR]

The aim of the present study was to investigate the effect and the underlying mechanism of the combined treatment of rhynchophylla total alkaloids (RTA) and sinapine thiocyanate for protection against a prothrombotic state (PTS) associated with the tumor necrosis factor-alpha (TNF-α)-induced inflammatory injury of vascular endothelial cells (VECs). A TNF-α-induced VEC inflammatory injury model was established, and cell morphology of VECs was evaluated using scanning electron microscopy. In addition, reverse transcription-quantitative polymerase chain reaction and western blot analysis were performed to examine the mRNA and protein expression of coagulation-related factors, including nuclear factor-κB (NF-κB), transforming growth factor-β1 (TGF-β1), tissue factor (TF), plasminogen activator inhibitor (PAI-1), protease-activation receptors (PAR-1) and protein kinase C (PKC-α) in VECs. Combined treatment with RTA and sinapine thiocyanate was demonstrated to reduce, to a varying extent, the mRNA and protein expression of NF-κB, TGF-β1, TF, PAR-1, PKC-α and PAI-1. Furthermore, combined treatment with RTA and sinapine thiocyanate was able to downregulate the expression of coagulation-related factors in injured VECs, thereby inhibiting the PTS induced by vascular endothelial injury. The underlying mechanism is partially associated with the TF-mediated activation of the thrombin-receptor signaling pathway that suppresses coagulation during inflammation and balances fibrinolysis in order to inhibit fibrin generation and deposition.