A Network Pharmacology Approach to Explore the Potential Mechanisms of Huangqin-Baishao Herb Pair in Treatment of Cancer.

BACKGROUND The aim of this study was to identify the bioactive ingredients of Huangqin-Baishao herb pair and to reveal its anti-cancer mechanisms through a pharmacology approach. MATERIAL AND METHODS Detailed information on compounds in the HQ-BS herb pair was obtained from the Traditional Chinese medicine systems pharmacology (TCMSP) and screened by the criteria of OB ≥30% and DL ≥0.18. A systematic drug targeting model (SysDT) was used for compound targets prediction, and then the targets were analyzed for Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. The protein-protein interaction (PPI) network of HQ-BS targets was constructed, after identifying core networks through Cytoscape plugins. RESULTS We found 47 bioactive compounds of HQ-BS and 107 human-derived targets. A compound target network and a target signal pathway network were constructed and used for topological analysis. Kaempferol, beta-sitosterol, stigmasterol, wogonin, and oroxylin-a were identified as core compounds and pathways in cancer. The calcium signaling pathway, PI3K-Akt signaling pathway, TNF signaling pathway, chemical carcinogenesis, estrogen signaling pathway, proteoglycans in cancer, HIF-1 signaling pathway, thyroid hormone signaling pathway, VEGF signaling pathway, small cell lung cancer, prostate cancer, colorectal cancer, NOD-like receptor signaling pathway, and T cell receptor signaling pathway were found to be potential signals of HQ-BS in treating cancer. Through PPI network analysis, TNF signaling pathway, tryptophan metabolism, proteoglycans in cancer, cell cycle, and chemical carcinogenesis sub-networks were obtained. CONCLUSIONS HQ-BS contains various bioactive compounds, including flavonoids, phytosterols, and other compounds, and these compounds can inhibit or activate multiple targets and pathways against cancer.

Separate and Combined Response to UV-B Radiation and Jasmonic Acid on Photosynthesis and Growth Characteristics of Scutellaria baicalensis.

The negative effects of enhanced ultraviolet-B (UV-B) on plant growth and development have been reported with many species. Considering the ability of jasmonic acid (JA) to improve plant stress tolerance, the hypothesis that JA pretreatment could alleviate the adverse effects of UV-B on S. baicalensis was tested in this study with photosynthesis and growth characteristics. The results showed that UV-B or JA alone both induced photosynthesis inhibition and decreased biomass in stems and leaves. However, the photosynthetic reduction caused by increased UV-B was mainly related to the effect of nonstomatal-limitation, while that of JA was a stomatal-limitation effect. JA pretreatment prior to UV-B could remit the photosynthetic inhibition via the recovery of chlorophyll content, stomatal conductance; and intercellular CO2 concentration (especially the maximum electron transport rate increase). Furthermore, the coaction of JA and enhanced UV-B alleviated some disadvantageous effects on the leaf and did not aggravate the growth damage induced by their separate actions.

[Impact of sulfur fumigation on quality of Scutellariae Radix].

Sulfur fumigation usually causes transformation of chemical components of traditional Chinese medicine (TCM). As for the process and storage of TCM, the sulfur fumigation has been no longer recommended by the government. This study is to explore the impact of the sulfur fumigation on the quality of Scutellariae Radix. Distillation combined with ion-chromatography was used to evaluate the residue of sulfur dioxide in 180 batches of Scutellariae Radix decoction pieces and 26 batches of Scutellariae Radix herbs. In addition, LC-MS combined with principal component analysis (PCA) was used to compare the changes in chemical components and flavonoid content in radix scutellariae before and after sulfur fumigation. As a result, 74 batches of Scutellariae Radix decoction pieces were determined to be unqualified on the basis of the residue limit of sulfur dioxide in Chinese Pharmacopoeia 2015 Vol. Ⅳ (≤150 mg•kg⁻¹), with a qualification rate of 35.9%. All of the unqualified samples were decoction pieces. The fingerprint and PCA indicates that flavonoid content of Scutellariae Radix had no change before and after sulfur fumigation, but flavonoid glycosides decreased after sulfur fumigation and baicalin decreased sharply. The flavonoid glycoside was hydrolyzed to flavone aglycone in the process of sulfur fumigation, which caused the decrease in content of falvonoid glycosides and had an impact on the quality of Scutellariae Radix. The decoction pieces of Scutellariae Radix were seriously fumigated with sulfur in the processing, suggesting the key regulatory point is the decoction pieces production process.

Autotetraploidy induced in nianmaohuangqin (Radix Scutellariae Viscidulae) with colchicine in vitro.

OBJECTIVE: To establish and optimize the propagation of Nianmaohuangqin (Radix Scutellariae Viscidulae) and induce and characterize polyploidy of Nianmaohuangqin (Radix Scutellariae Viscidulae). METHODS: Buds from germinating seed-derived explants were induced by tissue culture. With an orthogonal test, different concentrations of 6-benzyl-aminopurine (BAP), indole-3-acetic acid (IAA) and kinetin (KT) were used to determine the optimal concentrations for the propagation of Nianmaohuangqin (Radix Scutellariae Viscidulae). The different concentrations of IAA and rooting powder (ABT) were used to induce rooting. A 0.3% w/v colchicine solution was used to induce polyploidy and the induced buds was identified by root-tip chromosome determination and stomatal apparatus observation. RESULTS: A large number of buds could be induced directly from epicotyl and hypocotyl explants on Murashige and Skoog (MS) (Murashige and Skoog 1962) medium supplemented with 1.1-1.3 mg/L BAP and 0.2 mg/L IAA. Root induction and development could be observed within 20 days of inoculation on 1/2 MS medium supplemented with 0.2 mg/L IAA and 0.1 mg/L ABT. Furthermore, 27 lines of autotetraploid individuals were obtained with a plantlet chromosome number of 2n = 4x = 36. CONCLUSION: Autotetraploid lines could be obtained through induction with colchicine in vitro, proving that this method might be used for plant selection and breeding.

[Study on rapid propagation of Scutellaria baicalensis variation].

OBJECTIVE: To establish and optimize the technology of taking root and promoting seedlings of white flower Scutellaria baicalensis test tube plantlet, and provide the theory and technology base for efficient factorization production system of white flower Scutellaria baicalensis. METHODS: Stem segments with axillary bud were cultured onto the different basic medium with different kinds and concentration of cytokinin and auxins to take root and produce strong seedling. RESULTS: The suitable culture medium for taking root of white flower Scutellaria baicalensis was 1/2 MS (all substance reduced half) + IBA 0.02 mg/L + sucrose 2%, the induction rate of root was 100%; The best medium for promoting seedling was 1/2 MS (all substance reduced half) + PP333 0.2 mg/L + IBA 0.02 mg/L + sucrose 2%, the seedling was green, the internode was normal, and its growth was vigorous and healthy. CONCLUSION: 1/2 MS (all substance reduced half) culture medium and relatively low concentration of sucrose is beneficial to inducing roots; Media adding appropriate concentration of IBA can significantly increase the root induction rate, the seedling has many stout roots; PP333 has dwarfing effect on seedlings and suitable concentration of PP333 can significantly improve the quality of the plantlets. A good technology of taking root and producing strong test tube plantlets is established.

[Effects of plant growth regulator mepiquat chloride on content of active compounds in Scutellaria baicalensis].

OBJECTIVE: To study the effect of plant growth regulator mepiquat chloride (DPC) on the growing development of plant and the content of active compounds in Scutellaria baicalensis. METHOD: After spraying DPC during the seedling period, the length of taproot, fresh weight of root, diameter of taproot and the length of stem were measured. The contents of baicalin, baicalein and wogonin were determined by HPLC, respectively. Total flavonoids and scavenging DPPH were determined with ultraviolet spectrophotometry. RESULT: After spraying DPC, fresh weight of S. baicalensis root was significant increased, and root diameter was also increased. The contents of baicalin and total flavonoids were significantly increased and baicalein and wogonin were decreased remarkably. Compared with controls, scavenging activity of ethanol extracts on DPPH free radical was no significant changed after spraying DPC. CONCLUSION: Plant growth regulator DPC could regulate the growth on the ground and underground effectively, and could enhance the content of flavonoids compounds of S. baicalensis.

[Effect on different concentrations of exogenous hormones on baicalin in Scutellaria baicalensis callus].

OBJECTIVE: To determine the content of baicalin in Scutellaria baicalensis callus induced by different doncentrations of exogenous hormones. METHOD: HPLC system was adopted to determine baicalin in S. baicalensis callus. Chromatographic conditions: ODS column was adopted, with methanol-water-phosphate (47: 53: 0.2) as the mobile phase. The flow velocity was 1 mL x min(-1), the detective wavelength was 280 nm, and the temperature of column was room temperature. RESULT: S. baicalensis callus induced by 6-BA 1.0 mg x L(-1) + NAA 0.5 mg x L(-1) showed the highest baicalin content, up to 49.78 mg x g(-1). CONCLUSION: The experiment is such a simple, rapid and stable method for determining the baicalin content that it can be used for determining the baicalin content in S. baicalensis callus.

[Effects of PEG stress on flavonoids accumulation and related gene expression in suspension of Scutellaria baicalensis].

OBJECTIVE: To study the effects of PEG stress on baicalin, baicalein accumulation induced by an increased concentration of PEG solution and the related genes' expression in suspension of Scutellaria baicalensis. METHOD: The content of baicalin, baicalein in suspension of S. baicalensis was determined by HPLC. The related genes' expression was analyzed by semi-quantitative PCR. RESULT: The content of proline in suspension of S. baicalensis was promoted by PEG treatment. Ten percent PEG treatment promoted the expression of PAL and the content of baicalein in experimental material via a drought stress. 20% PEG solution treatment promoted the expression of UBGAT. At the same time, the increased activity of APX inhibited the progress of eliminating reactive oxygen by baicalein, which induced the transformation from baicalein to baicalin. CONCLUSION: Active ingredient in suspension of S. baicalensis was promoted significantly via a stress of light concentration of PEG solution.

[Correlation between H2O2 scavenging system and flavonoids accumulation of Scutellaria baicalensis].

OBJECTIVE: Study on correlation between H2O2 scavenging system and flavonoids accumulation of Scutellaria baicalensis. METHOD: The content of baicalin and baicalein in suspension cell of S. baicalensis was determined by HPLC. The content of total flavonoids and H2O2, the activity of POD and PAL was detected by UV spectrophotometry. RESULT: The content of total flavonoid and the activity of PAL increased significantly in 12 days after 40 degrees C, dark and PEG stress. Around 12 days after NPA, NPA +40 degrees C, 40 degrees C, NPA + dark, dark and PEG stress, the content of baicalin declined and the content of baicalein rise, the activity of POD showed an increasing trend, and level of H2O2 remain stable. CONCLUSION: Moderate environmental stress could promote the accumulation of total flavonoids in S. baicalensis, baicalin convert to baicalein by POD, and maintaining the stability of H2O2 content to avoid oxidative damage.

Complexity of active medicinal ingredients in radix scutellariae with sodium hydrosulfite exposure.

Both plants and animals are living things made up of similar cells as well as organelles, and their essence of life is the same. However, plants face more environmental stress than animals and generate excessive reactive oxygen species (ROS), a group of small molecules that can harm proteins, necessitating distinctive metabolic processes. Secondary metabolites in plants are a group of chemical components that can eliminate ROS and can also exhibit medicinal properties; therefore, herbal medicines are often closely linked to the ecological significance of secondary metabolites. Why plants contain so many, not few, active medicinal ingredients is unknown. The root of Scutellaria baicalensis, a popular herbal medicine, is rich in various flavonoids with diverse structural features. Sodium hydrosulfite (Na2S2O4) can produce O˙-2 radicals and induce physical conditions under environmental stress. Using UHPLC-ESI-Q-TOF-MS/MS analysis, a total of 25 different compounds were identified in the roots of S. baicalensis between the Na2S2O4 groups and suitable conditions. Based on the results of the t-test (P<0.05) performed for the groups and ions with values of VIP ≥ 2, the most significantly different chemical markers with Na2S2O4 treatment were shikimic acid, citric acid, baicalin, wogonoside, baicalein, wogonin, 3,5,7,2',6'-pentahydroxyflavanone, 5,2',6'-trihydroxy-7,8-dimethoxy flavone, chrysin, eriodictyol, 5,8-dihydroxy-6,7 -dimethoxy flavone, skullcapflavone Ⅱ, and 5,7-dihydroxy-6,8,2',3'-tetrame thoxyflavone, and most of them were free flavonoids with many phenolic hydroxyl or methoxyl groups and characteristically high antioxidant activities. S. baicalensis roots modified their ability to eliminate ROS and maintained the equilibrium of ROS through the multitudinous biosynthesis and conversion of flavonoids, which is similar to the equilibrium established by an intricate buffer solution and perfectly explains the diversity and complexity of medicinal plant ingredients.

[Effects of N P K on yield and baicalin contents of Scutellaria baicalensis].

OBJECTIVE: To find out the effects of different levels and combinations of nitrogen, phosphorus and potassium on the growth and secondary metabolites of Scutellaria baicalensis. METHODS: The quantities of growth, physiological indicators and baicalin contents in different fertilization treatment were calculated and compared. RESULTS: The growth physiological indicators and baicalin contents of S. baicalensis in complex fertilization treatment were better than those in single element fertilization treatment, and nutrition balance types better than maladjustment type. The reasonebly fertilitzing in complex fertilization treatment, nitrogen could improve the fertilizer efficiency of phosphorus. CONCLUSION: According to the fertilization mathematic model, the optimum fertilization dosage should be primarily defined as N: 22.6-25.0 g/m2, P: 30.6-44.6 g/m2, K: 5.4-21.4 g/m2, that is 2N:3P:1K.

[Effects of three fungicides on arbuscular mycorrhizal fungal infection and growth of Scutellaria baicalensis Georgi].

To make full use of arbuscular mycorrhizal (AM) fungal resources to increase the quantity and quality of Chinese medicinal materials and reduce the use of fungicide and environmental pollution, this research was carried out under pot culture condition to study the effects of benomyl, difenconazole and flusilazol on AM fungal (Glomus mosseae) colonization and the growth of Scutellaria baicalensis. The results showed that Glomus mosseae generally promoted the growth of host plant under the same fungicide application, and the effect of Glomus mosseae was different with different fungicides. The growth of non-mycorrhizal Scutellaria baicalensis was inhibited by applying with different fungicides. On mycorrhizal plants, contents of K in shoot and Fe in root significantly decreased under benomyl treatment; the contents of total N, K in shoot, Baicalin and Ca in root significantly decreased under difenconazole treatment; mycorrhizal colonization, the contents of total P, Baicalin, K and Cu in shoot, total N, Ca, Zn and Fe in root significantly decreased under flusilazol. The inhibitory effects of flusilazol on the Glomus mosseae colonization and growth of Scustellaria baicalensis were greater than those of difenconazole and benomyl. It is reasonable to choose less toxic fungicides in Scutellaria baicalensis cultivation to reduce the harmful effects on arbuscular mycorrhizal fungi.

Jasmonic acid (JA) acts as a signal molecule in LaCl(3)-induced baicalin synthesis in Scutellaria baicalensis seedlings.

Rare earth elements (REEs) have been widely used to increase accumulation of biomass and secondary metabolites in medicinal plants in China. However, very few studies have investigated how REEs mediate secondary metabolism synthesis in medicinal plants. Lanthanum (La), an important REE, is known to improve the accumulation of secondary metabolites in medicinal plants and is widely distributed in China. However, few studies have evaluated the signal transduction leading to La-induced secondary metabolism in medicinal plants. In this study, LaCl(3) treatment-induced multiple responses in Scutellaria baicalensis seedlings, including the rapid generation of jasmonic acid (JA), sequentially followed by the enhancement of baicalin production. Direct application of JA also promoted the synthesis of baicalin in the absence of LaCl(3). LaCl(3)-induced baicalin synthesis was blocked by two different JA synthesis inhibitors. Our results showed that JA acts as a signal component within the signaling system leading to La-induced baicalin synthesis in S. baicalensis seedlings.

Arsenic accumulation in Scutellaria baicalensis Georgi and its effects on plant growth and pharmaceutical components.

Scutellaria baicalensis Georgi is a traditional Chinese medicinal plant. The effects of arsenic (As) on the growth and the formation of pharmaceutical components of S. baicalensis, and the uptake and accumulation of As by S. baicalensis were investigated using a field pot-culture experiment. The results show that spiking low concentrations of As (