Exploring the mechanism of anti-hereditary Parkinson's disease of baicalein based on PINK1 RNAi Drosophila model / 药学学报

The aim of this study was to investigate the effect of baicalein on a Drosophila model of hereditary Parkinson's disease caused by gene mutations and to preliminarily elucidate the mechanism of baicalein in delaying hereditary Parkinson's disease. In this paper, PTEN-induced putative kinase 1 (PINK1)-RNAi Parkinson's Drosophila were used as the model group and wild-type Drosophila w1118 were used as the control group. Different doses of baicalein and Madopa were administered to the model group to observe their effects on the life span, motor ability, the abnormal rate of wings, dopamine content and dopaminergic neurons of PINK1-RNAi Parkinson's Drosophila and their effects on mitochondrial dysfunction including adenosine triphosphate (ATP), mitochondrial DNA (mtDNA) and reactive oxygen species (ROS) content. The results showed that the effective administration doses of baicalein were 0.8 mg·mL-1 for low concentration, 1.6 mg·mL-1 for medium concentration and 3.2 mg·mL-1 for high concentration, and the optimal administration dose of the positive drug Madopa was 0.1 μg·mL-1. Baicalein and Madopa could significantly improve the life span, exercise ability and reduce the abnormal rate of wings of PINK1-RNAi male Drosophila (P < 0.05), and low dose baicalein showed the best effect; baicalein could improve the loss of dopaminergic neurons, and the effects of low dose and high dose were the best, but Madopa showed no significant effect; baicalein and Madopa had no significant effect on dopamine content (P > 0.05). Baicalein and Madopa could increase the ATP content of PINK1-RNAi male Drosophila (P < 0.05), and low dose baicalein showed the best effect; middle dose baicalein could significantly increase the mtDNA content of PINK1-RNAi male Drosophila (P < 0.05), but Madopa had no significant effect; baicalein and Madopa had no significant effect on ROS content (P > 0.05).

Protective effect and mechanism of baicalein on hypoxia-induced cortical neuron injury in rats / 中国药房

OBJECTIVE To explore the protective effect and possible mechanism of baicalein on hypoxia-induced cortical neuron injury in rats. METHODS The cortical neurons of rats （RN-C cells） were studied and cultured under hypoxic conditions （5%CO2， 94% N2， 1%O2） for 24 hours； the effects of different concentrations of baicalein （0.01， 0.1， 1， 10， 100 μmol/L） on the survival rate of hypoxic RN-C cells were investigated； the effects of baicalein （0.1 μmol/L） on the activities of lactate dehydrogenase （LDH） and superoxide dismutase （SOD）， the content of malondialdehyde （MDA）， migration rate， apoptotic rate， cell cycle and the expressions of cleaved caspase-3， B-cell lymphoma-2 （Bcl-2） and Bcl-2 X protein （Bax） were all detected. RESULTS Compared with control group， the survival rate of cells in the hypoxia group was significantly reduced （P＜0.01）； 0.01， 0.1 and 1 μmol/L baicalein could reverse survival rate of hypoxia-induced cortical neurons （P＜0.05 or P＜0.01）. Scratch experiments showed that baicalein significantly increased the migration rate of hypoxic RN-C cells （P＜0.01）. Compared with control group， the activity of LDH in the supernatant and the content of MDA in the cells， apoptotic rate and the proportion of cells in G1 phase， were significantly increased in the hypoxia group， while SOD activity and the proportion of cells in G2+S phase was decreased significantly （P＜0.01）. The protein expressions of cleaved caspase-3 were increased significantly， while the ratio of Bcl-2/Bax in cells was significantly reduced （P＜0.05 or P＜0.01）. Compared with hypoxia group， the above indexes were all reversed significantly in baicalein group （P＜0.01）. CONCLUSIONS Baicalein can promote the proliferation and migration of cortical neurons， improve hypoxia-induced cell apoptosis and cell cycle distribution， decrease the activity of LDH in supernatant and the level of cellular lipid peroxidation， and improve antioxidant levels. Its mechanism may be related to regulating the caspase- 3/Bax/Bcl-2 pathway.

Mechanism of Baicalein in Inhibiting Microglial Activation and Protecting SH-SY5Y Nerve Cells / 中国实验方剂学杂志

ObjectiveTo investigate the effect of baicalein （BAI） on SH-SY5Y cell injury in lipopolysaccharide （LPS）-activated BV-2 cells conditioned medium and its mechanism. MethodThe BV-2 cells were activated with 1 mg∙L-1 of LPS to establish the conditioned medium of the LPS group， and a blank group and groups of BAI with low， medium， and high concentrations （4， 8， 16 μmol∙L-1） were established. SH-SY5Y cells were cultured with the conditioned medium of each group. The cell viability of BV-2 cells in each group after the intervention was determined by cell counting kit-8 （CCK-8）. The content of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β） in the supernatant of BV-2 cells in each group was determined by enzyme-linked immunosorbent assay （ELISA）. The protein expression of α-synuclein （α-syn） and tyrosine hydroxylase （TH） in SH-SY5Y cells was observed by immunohistochemical （IHC） staining， and the nuclear transfer of nuclear factor kappa-B p65 protein （NF-κB p65， p65） in SH-SY5Y cells was observed by immunofluorescence （IF）. The protein expression of Toll-like receptor 4（TLR4）， p65， phosphorylated p65 （p-p65）， and Myeloid differentiation factor 88 （MyD88） in SH-SY5Y cells was observed by Western blot. ResultAs compared with the blank group， the viability of BV-2 cells in the LPS group was significantly decreased （P<0.01）， and the content of TNF-α， IL-6， and IL-1β in the cell supernatant was significantly increased （P<0.01）. As compared with the LPS group， the cell viability was significantly increased in groups of BAI with low， medium， and high concentrations （P<0.01）， and TNF-α in the cell supernatant was significantly decreased （P<0.01）. The content of IL-6 in the cell supernatant was decreased in the BAI group with high concentration （P<0.05）， and the content of IL-1β in the cell supernatant was significantly decreased in the BAI groups with medium and high concentrations （P<0.01）. The results of conditioned medium cultured SH-SY5Y cells showed that as compared with the blank group， the protein expression of p65 in the LPS group entered into the nucleus and accumulated， and the protein expression of TH was significantly decreased （P<0.01）， while that of α-syn， TLR4， MyD88， and p-p65 was increased （P<0.05， P<0.01）. Compared with the LPS group， the protein expression of p65 in SH-SY5Y cells in BAI groups with low， medium， and high concentrations gradually dispersed into the cytoplasm and had the enhanced protein expression of TH （P<0.01） but the lowered protein expression of α-syn （P<0.01）. The protein expression of TLR4， MyD88， and p-p65 was decreased in the BAI group with high concentration （P<0.05， P<0.01）， the protein expression of p-p65 and MyD88 was decreased in the BAI group with medium concentration， and the protein expression of MyD88 was decreased in the BAI group with low concentration （P<0.05）. There was no significant difference in the protein expression of p65 among groups. ConclusionBAI can inhibit the activation of BV-2 cells， thereby inhibiting the inflammatory response caused by LPS and further inhibiting the damage of inflammation to SH-SY5Y cells. The mechanism may be related to the regulation of the TLR4/MyD88/NF-κB signaling pathway and reduction of the inflammatory response， thus playing a neuroprotective role.

Effect of baicalein combined with 5-FU on proliferation, apoptosis and migration of H446 cells by regulating ROS/MAPK signaling pathway / 中华内分泌外科杂志

Objective:To investigate the effects of baicalein combined with 5-FU on the proliferation, apoptosis and migration of small cell lung cancer H466 cells, and further to explore its sensitization mechanism.Methods:H466 cells were cultured in vitro and divided into blank control group, baicalein single treatment group, 5-FU single treatment group and combined treatment group. CCK-8 experiment was used to detect cell survival rate in each group; Flow cytometry was used to detect cell apoptosis and changes in ROS levels; Western blot was used to detect changes in the protein expression of the MAPK pathway.Results:CCK-8 results showed that the survival rates of the four groups of cells after 24 h treatment were 114.3% ± 2.8%, 79.8% ± 3.4%, 57.6% ± 1.8%, and 40.3% ± 2.7%. Compared with the other three groups, the combined treatment group had stronger inhibitory effect on the proliferation of H446 cells, and the difference was statistically significant ( P<0.001) . The proportion of apoptotic H446 cells in the combined treatment group was 49.2%±3.8%, which was significantly different from 33.9%±4.3% in the 5-FU single treatment group, and the difference was statistically significant ( P<0.001) . The inhibition rate of H446 cell migration in the combined treatment group was higher than that in the 5-FU single treatment group, and the difference was statistically significant ( P <0.001) . Combined treatment of baicalein and 5-FU can up-regulate ROS levels in H446 cells, thereby regulating the MAPK signaling pathway. Conclusion:The combined use of baicalein and 5-FU can increase the level of ROS in lung cancer H446 cells, activate the JNK and p38 signaling pathways, inhibit the ERK signaling pathway, and enhance 5-FU's proliferation inhibition, apoptosis induction and migration inhibition effects on H446 cells, which provides a certain experimental basis for the application of baicalein in small cell lung cancer.

Effect of Acute Liver Injury on Pharmacokinetics and Tissue Distribution of Baicalin Magnesium in Rats / 中国实验方剂学杂志

ObjectiveTo establish a high performance liquid chromatography （HPLC） for simultaneous determination of baicalin magnesium and baicalein in rat plasma and tissues， and to investigate the effect of acute liver injury on pharmacokinetics and tissue distribution of baicalin magnesium in rats. MethodAcute liver injury rat model was induced by carbon tetrachloride （CCl4）. Normal rats and acute liver injury model rats were given an equal dose （287.31 mg·kg-1） of baicalin magnesium aqueous solution by intragastric administration， the orbital blood was collected at different time points， and HPLC was used to simultaneously determine the concentrations of baicalin magnesium and baicalein in rat plasma at each time point， the concentration-time curves were drawn， the pharmacokinetic parameters were calculated with DAS 3.0， and SPSS 23.0 was used for statistical analysis. After oral administration of baicalin magnesium aqueous solution， HPLC was used to simultaneously determine the contents of baicalin magnesium and baicalein in rat liver， lung， kidney， stomach， brain and small intestine at different time points， the mobile phase was 0.1% phosphoric acid aqueous solution-methanol， and the detection wavelength was 278 nm. ResultIn the acute liver injury model group， the peak concentration （Cmax） of baicalin magnesium was 0.58 times that of the normal group， the area under concentration-time curve （AUC0-t） was 0.5 times that of the normal group （P<0.05）， the apparent volume of distribution （Vd） was 2.3 times that of the normal group （P<0.05）， and baicalein is almost undetectable in plasma. The content of baicalin magnesium in liver， stomach and brain of the acute liver injury model group was higher than that of the normal group at each time point， while the content of baicalin magnesium in the samples of lung at 8 h， kidney at 8 h and 12 h， and small intestine at 0.333 h was lower than that of the normal group. The content of baicalein in lung， stomach and small intestine of the model group was higher than that of the normal group at each time point， while the content of baicalein in the tissue samples of liver at 6， 8 h and kidney at 0.333， 4， 6 h was lower than that in the normal group， and baicalein could hardly be detected in the brain. ConclusionAfter intragastric administration of the same dose of baicalin magnesium aqueous solution， acute liver injury induced by CCl4 can affect the pharmacokinetics and tissue distribution characteristics of baicalin magnesium in rats， and there is biotransformation of baicalin magnesium and baicalein in liver， lung， kidney， stomach and small intestine.

Baicalein Mediated FAK Protein Regulates PI3K/Akt Signaling Pathway to Inhibit Proliferation and Migration of Gastric Cancer HGC-27 Cells / 中国实验方剂学杂志

ObjectiveTo study the possible molecular mechanism of baicalein （BAI）-mediated focal adhesion kinase （FAK） in the regulation of phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway to inhibit the proliferation and migration of gastric cancer HGC-27 cells. MethodThe gastric epithelial GES-1 cells and gastric cancer HGC-27 cells were respectively treated with BAI （0， 5， 15， 25， and 50 μmol·L-1） for 48 h， and then methyl thiazolyl tetrazolium （MTT） assay was adopted to detect effect of BAI on cell proliferation. Western blot （WB） was employed to detect the expression of FAK and the proteins related to epithelial-mesenchymal transition （EMT） and PI3K signaling pathway after intervention with different concentrations of BAI. The HGC-27 cells stably overexpressing FAK were constructed with lentivirus-mediated transfection technique， and the transfection of FAK was detected through WB and green fluorescent protein （GFP）. The cells were divided into empty vector （NC） group， BAI group， FAK overexpression group， and BAI-treated FAK overexpression group， and cell proliferation activity was detected by MTT assay. The colony formation and cell migration were observed via colony formation assay and Transwell migration assay， respectively. The expression of proteins involved in EMT and PI3K signaling pathways were detected by Western blot. ResultCompared with the NC group， BAI （15， 25 and 50 μmol·L-1） inhibited the proliferation of HGC-27 cells in a dose-dependent manner （P<0.05， P<0.01） while did not affect that of GES-1 cells. BAI （5， 15 and 25 μmol·L-1） down-regulated the expression level of p-FAK （P<0.05， P<0.01）. Compared with NC group， FAK overexpression group showed up-regulated expression level of FAK in HGC-27 cells. The HGC-27 cells in both NC group and FAK overexpression group had green fluorescence. Compared with NC group， BAI inhibited the growth， colony formation， and migration， while FAK overexpression promoted those of HGC-27 cells. The treatment of FAK overexpression group with BAI inhibited the enhancement of cell proliferation and migration （P<0.05）. WB showed that compared with NC group， BAI （15， 25 μmol·L-1） significantly up-regulated the expression of E-cadherin protein and down-regulated that of Vimentin， Snail， p-PI3K， and p-Akt protein in HGC-27 cells （P<0.05， P<0.01）. Compared with NC group， FAK overexpression group showed down-regulated expression of E-cadherin， up-regulated expression of p-FAK， Vimentin， and Snail， and increased ratios of p-FAK/FAK， p-PI3K/PI3K and p-Akt/Akt （P<0.05）. This phenomenon would be reversed after BAI treatment. ConclusionBAI can affect the proliferation and migration of gastric cancer HGC-27 cells by mediating FAK to regulate PI3K/Akt signaling pathway.

Study on mechanism difference of baicalein and wogonin inhibiting energy metabolism of hepatoma cells / 中国药房

OBJECTIVE To explore the difference in th e mechanis m of baicalein and wogonin inhibiting the energy metabolism of hepatoma cells. METHODS Human hepatoma HepG 2 cells were divided into blank control group （without medicine），different dose groups of baicalein and wogonin （1.25，2.5，5，10 and 20 μmol/L）. The effects of baicalein and wogonin on the viability of HepG 2 cells were detected by MTT assay. HepG 2 cells were divided into blank control group （without medicine），baicalein group and wogonin group. After administration ，the concentration of ATP in cell was detected by enhanced ATP kit. The levels of cell glycolysis and mitochondrial energy metabolism were evaluated by glycolysis and mitochondrial pressure test kit ；the affinity of baicalein and wogonin with key enzymes of energy metabolism was predicted by molecular docking ，and the key enzymes of energy metabolism with high affinity were screened ；the expression of key enzymes of energy metabolism was detected by Western blot. RESULTS Within the dose range of 2.5-20 μmol/L，the half inhibitory concentrations of baicalein and wogonin were 12.84 and 24.09 μmol/L；baicalein 1.25 μmol/L and wogonin 2.5 μmol/L had no effect on cell viability ，so it was selected as the dosage for subsequent experiments. Compared with blank control group ，the concentration of ATP in HepG 2 cells decreased significantly in baicalein group and wogonin group （P＜0.05）；the inhibitory effects on basic acidification rate of HepG 2 cells in wogonin group were significantly stronger than those of baicalein group （P＜0.05），but there was no significant difference between them on the basic oxygen consumption rate （P＞0.05）；baicalein had strong binding to pyruvate kinase M 2 and mitochondrial enzyme complexes Ⅰ（CⅠ），C Ⅱ and C Ⅳ，while wogonin only had strong binding to pyruvate kinase M 2； wogonin could significantly down-regulate the protein expressions of hexokinase ，phosphofructokinase，pyruvate kinase M 2，CⅠ， C Ⅱ and C Ⅳ（P＜0.05），but there was no statistical significance in the effect of baicalein on the regulation of these enzymes （P＞ 0.05）. CONCLUSIONS Both baicalein and wogonin can inhibit the energy metabolism of hepatoma HepG 2 cells，but the mechanism is different ：the effect of baicalein is related to the activity of key enzymes ，while the effect of wogonin is related to the inhibition of the expression of key enzymes of energy metabolism.

Baicalein inhibits neuroinflammation via NOX2/STAT1/NF-κB pathway in LPS-induced BV-2 cells based on bioinformatics methods / 药学学报

This study identified the exact molecular mechanisms of baicalein on neuroinflammation in lipopolysaccharide (LPS)-induced BV-2 cells. Bioinformatics methods and molecular docking were integrated for predicting the potential targets and mechanisms of baicalein. Immunofluorescence staining and Western blot were used to analyze the predicted key targets [inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2)], the expression level of protein related to signal transducer and activator of transcription 1/nuclear factor kappa-B (STAT1/NF-‍κB) signaling pathway and its upstream regulator NADPH oxidase-2 (NOX2), and then the mechanism of baicalein in alleviating neuroinflammation was explored. The results showed that iNOS and COX-2 were predicted as the key targets and NF-κB signaling pathway was one of the important pathways by bioinformatics methods and molecular docking. Experimental verification showed that baicalein could significantly reduce the expression of iNOS and COX-2, inhibit the phosphorylation of NF-κB and STAT1 and the production of NOX2 in LPS-induced BV-2 cells. To sum up, baicalein could effectively inhibit the inflammatory reaction in LPS-induced BV-2 cells through regulating NOX2 (gp91phox/p47phox)/STAT1/NF-κB pathway.

Effect of baicalein on the expression of glutamate receptor related protein in PC12 cells damaged by Aβ 25-35 / 国际中医中药杂志

Objective:To study the effect of baicalein on the expression of glutamate receptor related protein in PC12 cells injured by Aβ 25-35. Methods:PC12 cells were divided into control group, model group, estradiol group and baicalein group with different concentrations. The survival condition of PC12 cells in each group were detected by thiazole blue (MTT). PC12 cells were divided into control group, model group, estradiol group and baicalein group. The control group and model group were cultured with DMEM medium, and the estradiol group was added with 1×10 -3 μmol/L estradiol DMEM medium, baicalein group was added with 1 μmol/L baicalein DMEM medium. After 2 hours of intervention, 20 μmol/L Aβ 25-35 was added to the model group, estradiol group and baicalein group with induced PC12 cell injury. After 22 hours of intervention, flow cytometry was used to detect the apoptosis of PC12 cells. The expression of estrogen receptor β (ER β), phosphorylated c-Jun N-terminal kinase (p-JNK/JNK) and ionic receptor N-methyl-D-aspartate receptor 1 (NMDAR1), glutamate receptor 2 (GluR2) and calcium/calmodulin dependent protein kinase Ⅱ (CaMK Ⅱ) were detected by Western blot. Results:Compared with model group, 1 μmol/L baicalein significantly increased the proliferation rate [(95.80±2.47)% vs. (64.34±3.84)%]. The apoptosis rate of PC12 cells[(7.83±0.67)% vs. (12.84±0.91)%] was significantly decreased in baicalein group ( P<0.01). The expression of NMDAR1 (0.582±0.012 vs. 0.352±0.012), GluR2(0.538±0.017 vs. 0.355±0.006), ER β (0.362±0.015 vs. 0.262±0.018) in baicalein group were significantly increased ( P<0.01), the expression of p-JNK/JNK (0.476±0.013 vs. 0.752±0.014) and CaMK Ⅱ(0.499±0.019 vs. 0.670±0.016) in baicalein group were significantly decreased ( P<0.01). Conclusions:Baicalein has a protective effect on PC12 cells injured by Aβ 25-35. Its mechanism may be related to the inhibition of p-JNK/JNK activity by activating ERβ and regulating the expression of glutamate receptor related protein.

Protective effect of baicalein on injured PC12 cell induced by Aβ and its mechanism / 国际中医中药杂志

Objective:To investigate the protective effect of baicalein on injured PC12 cell induced by Aβ and explore its mechanism.Methods:The method of MTT was used to detect the cell activity of each group and screened the concentration of baicalein. The PC12 cells were randomly divided into the blank group, the Aβ group, the baicalin group and the estradiol group. 24 hours after inoculation, baicalein group was intervened with 1×10 -6 mol/L baicalein solution, and estradiol group was intervened with 1×10 -5 mol/L estradiol solution. Two hours later, except the blank group, the other groups were added with 1.5×10 -4 mol/L Aβ to make the model. MTT assay was used to detect the cell viability of each group after 24 hours of cultivation. Then used oxidation kit to detect the contents of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and lactate dehydrogenase (LDH) in each group. And the level of caspase-3 mRNA was detected by Real-time Quantitative PCR (RT-PCR). Then the Western blot method was used to detect the expressions of p-PI3K, p-AKT and caspase-3. Results:Compared with the Aβ group, the PC12 cell viability [(96.348 ± 0.571)%, (97.183 ± 0.714)% vs. (86.922 ± 0.429)%] in the baicalin group and the estradiol group significantly increased( P<0.01). The activities of SOD [(54.31 ± 1.34) U/mgprot, (57.38 ± 2.25) U/mgprot vs. (36.18 ± 2.24) U/mgprot] and GSH-PX [(4.46 ± 0.23) U/mgprot, (4.72 ± 0.31) U/mgprot vs. (2.05 ± 0.37) U/mgprot] significantly increased, and the level of LDH [(85.43 ± 0.92) nmol/ml, (82.46 ± 0.27) nmol/ml vs. (99.17 ± 0.52) nmol/ml] significantly decreased ( P<0.01). The expression of caspase-3 mRNA (2.24 ± 0.64, 2.33 ± 0.75 vs. 3.46 ± 0.46) and p-PI3K (0.46 ± 0.03, 0.44 ± 0.06 vs. 0.66 ± 0.09), p-AKT (0.43 ± 0.05, 0.41 ± 0.02 vs. 0.58 ± 0.03), caspase-3 (0.61 ± 0.03, 0.56 ± 0.53 vs. 0.92 ± 0.07) protein significantly decreased ( P<0.01). Conclusion:Baicalein could slow down cell apoptosis and oxidative reaction, reduce the damage of Aβ to PC12 cells by inhibiting the expression of PI3K/AKT pathway.

New advances in baicalein's antitumor effects and mechanisms / 药学实践杂志

Baicalein (BE) is an active ingredient derived from the root of Scutellaria baicalensis Georgi. It is a polyhydroxyflavonoid in structure and has many biological activities. Among them, the antitumor effects of baicalein have received widespread attention. It exerts anti-tumor effects by inducing tumor cell apoptosis and invasion, inhibiting tumor angiogenesis, and eliminating free radicals. This article reviews the most recent research works of baicalein in its anti-tumor effects and mechanisms. It is aimed to provide a theoretical basis for the search and development of potential new anti-tumor drugs.

Effect of molecular weight of polyethylene glycol on pharmacokinetics of baicalin / 药学学报

The aim of this study was to investigate the effects of polyethylene glycol (PEGs) with different molecular weights (MW: 400, 1 000, 4 000) on the pharmacokinetics of baicalin, and preliminarily analyze its mechanism. Rats were gavaged with baicalin (168 mg·kg-1) + aqueous solution or baicalin + PEGs solution and plasma samples were collected from 0 to 24 h after administration. The concentration of baicalin and its main metabolite baicalein 6-O-β-D-glucuronide (B6G) were determined at different time points by UPLC-MS/MS, and the pharmacokinetic parameters were calculated with DAS 3.0 software. The results showed that PEGs with different molecular weights could effectively increase the AUC0-t of baicalin and B6G, increase the Cmax, and prolong the t1/2, effectively increasing the concentration of baicalin and B6G in vivo. The mechanism may be by promoting the activity of uridine diphosphate glucuronosyl-transferases 1A8 (UGT1A8) and 1A9 (UGT1A9), thereby increasing the transformation rate of baicalin and B6G. The rate of metabolism of B6G was faster than that of baicalin, suggesting that PEGs had a higher affinity for UGT1A8, and PEG400 had the most significant effect. The purpose of this study was to provide a basis for the clinical safe use of baicalin and other flavonoids and the design of new dosage forms with the participation of PEGs. The animal experiment protocol in this study was approved by the Experimental Animal Ethics Committee of Guizhou Medical University.

Baicalein reduces Aβ25-35-induced PC12 cell damage by inhibiting the JAK2/STAT1 pathway / 药学学报

This study investigated the mechanism by which baicalein protected PC12 cells from Aβ25-35-induced injury. PC12 cells were treated with Aβ25-35 (20 μmol·L-1) and the ability of baicalein to prevent apoptosis was investigated by monitoring changes in cell morphology, Hoechst 33342 staining, and measurement of inflammatory factors. Western blotting was used to detect the expression of the apoptosis-related proteins cysteinyl aspartate specific proteinase-3 (caspase-3), cleaved cysteinyl aspartate specific proteinase-3 (cleaved caspase-3), proteins involved in the Janus kinase 2/signal transducer and activator of transcription 1 (JAK2/STAT1) pathway, and downstream inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). The results show that baicalein (80 μmol·L-1) can significantly inhibit apoptosis and the release of inflammatory factor IL-8 and TNF-α in Aβ25-35-treated PC12 cells. Western blotting results showed that baicalein can inhibit the phosphorylation of JAK2 and STAT1 and decrease the expression of downstream iNOS and COX-2, thereby inhibiting the JAK2/STAT1 signaling pathway and preventing Aβ25-35-induced PC12 cell damage.

Enhanced dissolution and eliminated gelation of lenvatinib mesylate by coamorphous system / 中国药科大学学报

@#Lenvatinib mesylate (LF), a multi-target tyrosinase inhibitor mainly used in the treatment of a variety of cancers, has low oral bioavailability mainly due to its gelation during the dissolution process. In the current study, in order to enhance dissolution and eliminate gelation of LF, a supramolecular coamorphous system of LF-baicalein (BAI) (molar ratio, 1∶1) was prepared by rotary evaporation and characterized by PLM, PXRD, DSC and FTIR. Results indicated the formation of coamorphous system with a single Tg of 118 °C. Different from original LF crystal, no gelation phenomenon was observed during the dissolution of coamorphous LF-BAI. In addition, the dissolution rate of LF was increased by 2.2-fold after coamorphization. Meanwhile, the dissolution rate of the co-former BAI was also enhanced by more than 25.4-fold. Stability test showed that the prepared coamorphous system had a good physical stability for at least 90 days under 25 °C/ 60%RH and 40 °C /75%RH conditions.

Mechanisms of baicalein inhibiting metastasis of esophageal squamous cell carcinoma through ERK/ELK-1/Snail signaling pathway / 药学学报

This study aims to investigate the effect of baicalein on the metastasis of esophageal squamous cell carcinoma (ESCC) cells, and to elucidate the potential molecular mechanisms. Wound healing and Transwell migration and invasion assays were performed to detect the effect of baicalein on the migration and invasion of EC9706 and KYSE30 cells; the nude mice models of lung metastasis were applied to examine the function of baicalein in metastasis of ESCC by using KYSE30 cells. All animals were received humane care according to the Institutional Animal Care Guidelines approved by the Experimental Animal Ethical Committee of Henan University. Western blot assay was used to detect the protein levels of ERK/ELK-1/Snail signaling pathway. The data showed that baicalein significantly inhibited the migration and invasion of EC9706 and KYSE30 cells; Mechanistically, baicalein treatment led to a dramatically reduced expression of phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2, T202/Y204), p-ETS-domain containing protein-1 (p-ELK-1, S383), Snail, N-cadherin, and Vimentin, and a statistical increase of E-cadherin expression in EC9706 and KYSE30 cells; Furthermore, the inhibition of ERK1/2 by U0126 or siRNA remarkably enhanced the effect of baicalein on the above proteins. In summary, baicalein probably inhibits the migration, invasion, and metastasis of ESCC cells via blocking the ERK/ELK-1/Snail signaling pathway.

Investigation of Effect of Polyethylene Glycol 400 on Pharmacokinetics and Anti-inflammatory Effect of Baicalin Based on UPLC-MS/MS and Molecular Docking Techniques / 中国实验方剂学杂志

Objective:To investigate the effects of polyethylene glycol 400 （PEG400） on the pharmacokinetics and anti-inflammatory effect of baicalin， and to preliminarily explore the anti-inflammatory effects of baicalin and its main metabolite baicalein 6-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucuronide （B6G） by molecular docking. Method:Rats were randomly divided into two groups with water and PEG400 as the dissolving matrix， and rats were administrated the equal dose of baicalin aqueous solution （baicalin+water group） and baicalin PEG400 solution （baicalin+PEG400 group）. After the plasma samples were processed at different time periods， the concentrations of baicalin and B6G in rat plasma were determined by ultra performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）， and pharmacokinetic parameters were processed by DAS 3.2.2 software. Mice were randomly divided into a blank group （normal saline， 20 mL·kg^-1）， aspirin group （dose of 0.2 g·kg^-1）， baicalin/baicalin+PEG400 high and low dose （3.0， 1.5 g·kg^-1） groups， after continuous administration for 7 days， the mouse ear swelling and foot swelling models were established， and the swelling degree and swelling inhibition rate were calculated. Result:The pharmacokinetic study showed that compared with baicalin+water group， the plasma concentrations of baicalin and B6G increased after administration of baicalin PEG400 solution， and the area under the curve （AUC_0-<italic>_t</italic>） increased by 2.36， 1.97 times， and the peak concentration （<italic>C</italic>_max） increased by 2.12， 1.65 times， respectively. The results of mouse ear and foot swelling inflammation models showed that the anti-inflammatory effect was enhanced after intragastric administration of baicalin PEG400 solution. In addition， molecular docking results showed that baicalin and B6G could site bind to multiple target proteins ［tumor necrosis factor （TNF）-<italic>α</italic>， interleukin （IL）-6， IL-1<italic>β</italic>， prostaglandin E₂ （PGE₂） and nuclear transcription factor-<italic>κ</italic>B （NF-<italic>κ</italic>B）］ with higher affinity， which was superior to the positive drug aspirin. Conclusion:PEG400 can increase the plasma concentration of baicalin and its main metabolite B6G， and enhance the anti-inflammatory effect. Baicalin and B6G can form strong hydrogen bonds with various inflammatory factors and of nuclear transcription factors， it is speculated that baicalin and B6G jointly play an anti-inflammatory role.

Baicalein Regulates Colony Formation of Triple-negative Breast Cancer Cells by Inhibiting YAP Entry into Nucleus / 中国实验方剂学杂志

Objective:This study aims to observe the effect of baicalein on the clonal formation of triple negative breast cancer MDA-MB-231 and MDA-MB-468 cells， and to explore the mediation role of Yes- related protein （YAP） in it. Method:MDA-MB-231 and MDA-MB-468 cells were treated with baicalein. Thiazole blue （MTT） colorimetric method was used to detect cell proliferation ability. Plate cloning experiments was used to detect the colony forming ability. Immunofluorescence method was used to detect the nuclear distribution of YAP， and Western blot test was used to detect the protein expression levels of YAP large tumor suppressor factor 1 （LATS1）， YAP， phosphorylated Yes- related protein（p-YAP） and phosphorylated YAP large tumor suppressor factor 1 （p-LATS1）. Result:Compared with the blank group， baicalein （40， 80， 160 μmol·L^-1） significantly inhibited the proliferation ability of MDA-MB-468 and MDA-MB-231 cells （<italic>P</italic><0.05， <italic>P</italic><0.01）， and the inhibitory effect was dose-dependent. The half inhibit concentration（IC₅₀） of baicalein against MDA-MB-468 and MDA-MB-231 cells were （80.3±7.2），（70.4±6.5） μmol·L^-1， respectively. Compared with blank group， baicalein （5， 10， 20 μmol·L^-1） had no significant effect on the proliferation of MDA-MB-468 and MDA-MB-231 cells， and the difference was not statistically significant. Compared with the blank group， baicalein （5， 10， 20 μmol·L^-1） significantly dose-dependently reduced the cell colony formation rates of MDA-MB-468 and MDA-MB-231 cells （<italic>P</italic><0.05， <italic>P</italic><0.01）， and baicalein （10， 20 μmol·L^-1） significantly inhibited the nuclear expression of YAP in MDA-MB-468 and MDA-MB-231 cells in a dose-dependent manner（<italic>P</italic><0.01）. Also， baicalin （5， 10， 20 μmol·L^-1） significantly up-regulated p-YAP and p-LATS1 protein expressions in MDA-MB-468 cells in a dose-dependent manner （<italic>P</italic><0.05， <italic>P</italic><0.01）. Baicalein （10， 20 μmol·L^-1） significantly up-regulated p-YAP and p-LATS1 protein expressions in MDA-MB-231 cells in a dose-dependent manner （<italic>P</italic><0.01）. Conclusion:Baicalein can inhibit colony formation of triple negative breast cancer MDA-MB-468 and MDA-MB-231 cells by mediating the reduction of YAP entry into the nucleus.

Stability Study of Baicalein-7-O-diglucoside / 中国药学杂志

OBJECTIVE: To investigate the stability of baicalein-7-O-diglucoside. METHODS: The chromatographic purity of baicalein-7-O-diglucoside and its solution in different condition was analyzed by HPLC-DAD. The degradation products of baicalein-7-O-diglucoside in its solvent were identified by LC-MS. Three degradation products in the solvent of baicalein-7-O-diglucoside were identified with applying electrospray ionization(ESI) source in the positive ion mode.RESULTS: ① Baicalein-7-O-diglucoside was stable with light and in 40, 80 and 105 ℃ within the inspection time in solid state. ②Baicalein-7-O-diglucoside in water was stable in room temperature and degrades quickly in 50%methanol and 50% ethanol. ③The degradation products of baicalein-7-O-diglucoside in solvent were 7-methoxy baicalein, 7-ethoxy baicalein and baicalein. The above research provides that baicalein-7-O-diglucoside is stable in solid state. Baicalein-7-O-diglucoside degrades in 50% methanol and 50% ethanol. CONCLUSION: Water should be used as the solvent for solution of baicalein-7-O-diglucoside. The solution of baicalein-7-O-diglucoside should be made up before use or stored in the refrigerator immediately after preparation.

Baicaleininhibitsthe biological behavior of hepatocellular carcinoma stemcells by downregulation of decoy receptor 3 expression / 中国组织工程研究

BACKGROUND: To date, there is no report on the anti-hepatoma mechanism of Chinese herbal medicine targeting hepatocellular carcinoma stem cells; therefore, relevant research is necessary. OBJECTIVE: To observe the effect of baicalein on the expression of Decoy receptor 3 in hepatocellular carcinoma stem cells as well as the biological behavior of hepatocellular carcinoma stem cells after down-regulation of Decoy receptor 3.METHODS: Hepatocellular carcinoma stem cells were obtained from hepatocellular carcinoma cell lines (purchased from the Cell Bank, Chinese Academy of Sciences, Shanghai, China), cultured and passaged. The cells were cultured in the low-glucose DMEM containing nothing (control group), 200 (high-dose group), 100 (middle-dose group) or 50 μmol/L (low-dose group) baicalein. After treatment with baicalein, RT-PCR and western blot were used to detect the expression of decoy receptor 3 at mRNA and protein levels. Cell counting kit-8, flow cytometry, and Transwell assay were used to detect the proliferation, cell cycle distribution, apoptosis and migration of hepatocellular carcinoma stem cells. RESULTS AND CONCLUSION: Compared with the control group, high-dose baicalein could significantly down-regulated the expression of decoy receptor 3 mRNA and protein in hepatocellular carcinoma stem cells. High-dose baicalein was then confirmed to be the best concentration of action. Compared with the control group, in the high-dose baicalein group, the proliferation ability of hepatocellular carcinoma stem cells decreased significantly, the percentage of S-phase and G2-phase cells increased, while the percentage of G1-phase cells decreased (P < 0.05). Compared with the control group, in the high-dose baicalein group, the apoptotic rate of hepatocellular carcinoma stem cells increased significantly, while the migration ability decreased markedly. To conclude, high-dose baicalein can inhibit a series of biological behaviors of hepatocellular carcinoma stem cells by down-regulating the expression of Decoy receptor 3, which provides an experimental basis for clinical treatment of hepatocellular carcinoma with Decoy receptor 3 as the target and hepatocellular carcinoma stem cells as the object.

Mechanism of Tanreqing Injection on treatment of coronavirus disease 2019 based on network pharmacology and molecular docking / 中草药

Objective: To study the mechanism of Tanreqing Injection (TRQI) on treatment of coronavirus disease 2019 (COVID-19) through network pharmacology and molecular docking, so as to provide theoretical basis for clinical treatment. Methods: The active compounds of TRQI were searched by literature, BATMAN-TCM, and TCMSP database. The potential targets of TRQI active compounds were searched by TCMSP. In Genecards database, “coronavirus” was used as the key word to search for coronavirus targets and the common targets were selected by mapping with TRQI. The network between the active compounds and common targets was established by Cytoscape 3.2.1. The common targets were imported into a STRING database for protein-protein interaction analysis, and the target protein interaction network diagram (PPI) was constructed. The key antiviral targets of TRQI were screened by combining two networks. “GlueGO 2.5.5” plug-in unit in Cytoscape 3.2.1 was used to perform GO biological process and KEGG pathway enrichment analysis. Results: A total of 54 components of TRQI were obtained and corresponding to 287 targets. Among them, there were 54 common targets and 34 key targets. GO analysis obtained 29 biological processes related to the treatment effect of TRQI, and KEGG analysis obtained 70 pathways. The results of molecular docking showed that kaempferol, quercetin, baicalein luteolin, and wogonin had good affinity with SARS-CoV-2 3CL hydrolase. Conclusion: The active compounds in TRQI may act as an antiviral agent by binding SARS-CoV-2 3CL hydrolase and regulating multiple signaling pathways. The molecular mechanism of TRQI in the treatment of COVID-19 indicated the synergistic features of multi-component, multi-target, and multi-pathway of traditional Chinese medicine, which provided an important scientific basis for further elucidating the mechanism of TRQI in the treatment of COVID-19.