Wogonoside induces its effects by modulating the expression and subcellular localization of Phospholipid scramblase 1 (PLSCR1) [5].

In RAW264.7 cells, wogonin (0-50 μM, 24 hours) suppresses the production of pro-inflammatory cytokines (IL-6 and TNF-α) and central mediators (NO, PGE2) generated by LPS [1]. Wogonin at 100 μM for 24 hours causes MDA-MB-231 and MCF-7 to proliferate [2]. Wogonin (100 μM, 9–24 hours) causes MDA-MB-231 cells to proliferate [2]. μM, 48-96 hours) can cause G1 phase induction and secretion, as well as suppress the growth of U937 and HL-60 cells [5].

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Wogonoside exhibited antiproliferative effects on U937 and HL-60 human leukemic cell lines and primary AML cells, as determined by trypan blue dye exclusion assays. Treatment with 50, 100, and 150 µM wogonoside for up to 5 days inhibited cell growth in a time- and dose-dependent manner [5].
In soft agar colony formation assays, treatment with 100 µM and 150 µM wogonoside for 4 days significantly reduced both the number and size of colonies formed by U937 and HL-60 cells [5].
Wogonoside induced G0/G1 phase cell cycle arrest in U937 and HL-60 cells in a dose-dependent manner after 48 hours of treatment, as shown by flow cytometry. This was associated with the downregulation of CDK4 and cyclin D1 protein levels, and the upregulation of the CDK4 inhibitor p16 (INK4A) [5].
Wogonoside induced differentiation in U937, HL-60, and primary AML cells. After 96 hours of treatment with 100 µM and 150 µM wogonoside, cells displayed morphological features of differentiation (e.g., lower nucleocytoplasmic ratio, chromatin condensation), increased NBT-reducing activity, and increased α-naphthyl acetate esterase activity. Flow cytometry analysis showed an increase in the percentage of cells expressing both CD11b and CD14, suggesting differentiation along the monocytic lineage [5].
Wogonoside (150 µM) increased PLSCR1 mRNA and protein levels in U937 and HL-60 cells through transcriptional activation, as this increase was blocked by the transcription inhibitor actinomycin D. It also downregulated c-Myc and upregulated p21waf1/cip1 protein expression [5].
Wogonoside promoted the trafficking of PLSCR1 into the nucleus, as demonstrated by western blotting of fractionated cells and immunofluorescence microscopy. It also facilitated the binding of nuclear PLSCR1 to the IP3R1 promoter, as shown by EMSA, and subsequently increased IP3R1 protein expression [5].
Transfection of U937 and HL-60 cells with PLSCR1 siRNA partially blocked the effects of wogonoside (150 µM), including the G0/G1 phase arrest, increased NBT-reducing activity, and upregulation of CD11b/CD14. It also inhibited the wogonoside-induced changes in p16, p21waf1/cip1, c-Myc, and IP3R1 protein levels [5].

In the MDA-MB-231 orthotopic model, wogonin (80 mg/kg, gavaged once every other day) inhibits the growth and metastasis of tumors [3]. In A549 cell xenograft models, wogonin (40 or 80 mg/kg, intraperitoneal injection, every three days) inhibits tumor growth and decreases Bcl-2/Bax distribution [4]. Wogonin can also inhibit the U937 xenograft model when administered intraperitoneally once daily for 14 days at a dose of 80 mg/kg.

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In a BALB/c nude mouse xenograft model with subcutaneously implanted U937 cells, treatment with wogonoside (80 mg/kg, i.p., every other day for 14 days) significantly suppressed tumor growth. At the end of treatment, the mean tumor volume in the treated group (2185±276 mm³) was 39% lower than in the control group (3559±421 mm³). The mean tumor weight was also reduced by 41% in the wogonoside-treated group (1287±340 mg) compared to the control group (2181±330 mg) [5].
In a NOD/SCID mouse model engrafted with primary human AML cells via tail vein injection, treatment with wogonoside (80 mg/kg, i.p., every other day for 14 days) significantly prolonged survival. The median survival duration was 14.5±2.7 days in the control group and 33.5±15.1 days in the wogonoside-treated group (P < .001, log-rank test) [5].

Western Blot Analysis[2]
Cell Types: MDA-MB-231 cells
Tested Concentrations: 100 μM
Incubation Duration: 9-24 hrs (hours)
Experimental Results: Increased expression of LC3-II and Beclin-1. Inhibits the phosphorylation levels of mTOR Ser2448 and p70S6K Ser-389.

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For proliferation assays, U937, HL-60, or primary AML cells were seeded at a density of 7×10⁴ cells/mL in 6-well plates and incubated with varying concentrations of wogonoside (0, 50, 100, 150 µM) for up to 5 days. Viable cell counts were performed daily using the trypan blue dye exclusion method with a hemocytometer [5].
For colony formation assays, cells were first treated with or without wogonoside (100, 150 µM) for 4 days. They were then cloned in soft agar and cultured for an additional 21 days. Colonies with a diameter >50 µm were counted under an inverted microscope [5].
For cell cycle analysis, cells treated with wogonoside (50, 100, 150 µM) for 48 hours were harvested, fixed, and stained with propidium iodide. The DNA content was then analyzed using a FACSCalibur flow cytometer, and the percentage of cells in each phase was quantified with Modfit software [5].
For differentiation analysis, cells were treated with wogonoside (100, 150 µM) or ATRA (1 µM) for 96 hours. Morphological assessment was done by Wright-Giemsa staining. NBT reduction was measured by incubating cells with NBT solution and counting the percentage of cells with purple-black formazan deposits. α-naphthyl acetate esterase activity was assessed using a commercial kit. Cell surface markers CD11b and CD14 were detected by flow cytometry after staining with specific antibodies [5].
For Western blot analysis, cells treated with wogonoside were lysed, and proteins were separated by SDS-PAGE, transferred to membranes, and probed with specific primary antibodies (e.g., against PLSCR1, CDK4, cyclin D1, p16, c-Myc, p21, IP3R1). Detection was performed using an Odyssey Infrared Imaging System [5].
For immunofluorescence, cells treated with 150 µM wogonoside for 48 hours were fixed, permeabilized, and incubated with anti-PLSCR1 primary antibody, followed by a FITC-labeled secondary antibody. Nuclei were counterstained with DAPI. Subcellular localization was visualized using confocal microscopy [5].
For quantitative real-time RT-PCR, total RNA was isolated using TRIzol reagent, and cDNA was synthesized. PLSCR1 mRNA levels were measured using SYBR Green PCR core reagents on an Applied Biosystems 7500 Fast Real-Time PCR System, with GAPDH as an internal control [5].
For the transcription inhibition study, cells were pretreated with 4 µg/mL actinomycin D for 4 hours, followed by 150 µM wogonoside for 48 hours. PLSCR1 protein expression was then analyzed by western blotting [5].
For EMSA, nuclear extracts from cells treated with 150 µM wogonoside for 48 hours were prepared. DNA binding was determined using an EMSA kit with double-stranded oligonucleotides containing the IP3R1 promoter sequence. For supershift experiments, an anti-PLSCR1 antibody was included in the reaction [5].
For siRNA transfection, U937 and HL-60 cells were transfected with PLSCR1 siRNA or non-specific control siRNA using Lipofectamine 2000 reagent according to the manufacturer's instructions. After transfection, cells were treated with or without 150 µM wogonoside for 96 hours and then subjected to further analysis [5].

Animal/Disease Models: MDA-MB-231 orthotopic model [3]
Doses: 80 mg/kg administered Method: po (oral gavage), once every other day.
Experimental Results: Inhibited tumor growth (46%) and metastasis in the brain, lungs, liver and bones. Increase the expression of E-cadherin at the original site and decrease the expression of MMP-9, vimentin and Twist1.

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For the U937 xenograft model, U937 cells suspended in Matrigel were injected subcutaneously into the bilateral flanks of BALB/c nude mice, creating two tumors per mouse. When tumors reached a palpable size (50–100 mm³), mice were randomized into two groups (n=5 per group). The treatment group received intraperitoneal injections of wogonoside at a dose of 80 mg/kg, while the control group received the solvent. Injections were administered every other day for a total of 14 days. Tumor volumes were measured every other day using calipers. At the end of the treatment period, mice were euthanized, and tumors were excised and weighed [5].
For the AML-bearing NOD/SCID mouse survival study, mice were first sublethally irradiated (2.4 Gy). Twenty-four hours later, primary human AML cells (2.5×10⁶ cells per mouse) were injected into the tail vein. Starting the next day, mice (n=6 per group) received intraperitoneal injections of wogonoside (80 mg/kg) or solvent every other day for 14 days. A blank animal group (no primary cells, treated with solvent) was also included. Animals were observed for 60 days post-cell injection, and survival was recorded [5].

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For the U937 xenograft model, U937 cells suspended in Matrigel were injected subcutaneously into the bilateral flanks of BALB/c nude mice, creating two tumors per mouse. When tumors reached a palpable size (50–100 mm³), mice were randomized into two groups (n=5 per group). The treatment group received intraperitoneal injections of wogonoside at a dose of 80 mg/kg, while the control group received the solvent. Injections were administered every other day for a total of 14 days. Tumor volumes were measured every other day using calipers. At the end of the treatment period, mice were euthanized, and tumors were excised and weighed [5].
For the AML-bearing NOD/SCID mouse survival study, mice were first sublethally irradiated (2.4 Gy). Twenty-four hours later, primary human AML cells (2.5×10⁶ cells per mouse) were injected into the tail vein. Starting the next day, mice (n=6 per group) received intraperitoneal injections of wogonoside (80 mg/kg) or solvent every other day for 14 days. A blank animal group (no primary cells, treated with solvent) was also included. Animals were observed for 60 days post-cell injection, and survival was recorded [5].

[1]. Wogonoside displays anti-inflammatory effects through modulating inflammatory mediator expression using RAW264.7 cells. J Ethnopharmacol. 2013 Jun 21;148(1):271-6.

[2]. Wogonoside induces autophagy in MDA-MB-231 cells by regulating MAPK-mTOR pathway. Food Chem Toxicol. 2013 Jan;51:53-60.

[3]. Wogonoside inhibits invasion and migration through suppressing TRAF2/4 expression in breast cancer. J Exp Clin Cancer Res. 2017 Aug 3;36(1):103.

[4]. Wogonoside induces apoptosis in human non-small cell lung cancer A549 cells by promoting mitochondria dysfunction. Biomed Pharmacother. 2018 Oct;106:593-598.

[5]. Wogonoside induces cell cycle arrest and differentiation by affecting expression and subcellular localization of PLSCR1 in AML cells. Blood. 2013 May 2;121(18):3682-91.

Baicalin 7-O-β-D-glucuronide is the 7-O-glucuronide of baicalin, belonging to the glycosyloxyflavones, monomethoxyflavones, monohydroxyflavones, monosaccharide derivatives, and β-D-glucuronic acid. It is functionally related to baicalin and is the conjugated acid of baicalin 7-O-β-D-glucuronic acid. Baicalin has been reported in Scutellaria indica, Scutellaria prostrata, and other organisms with relevant data. See also: Glycyrrhiza glabra (partial).

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Wogonoside is the main flavonoid component derived from the root of Scutellaria baicalensis Georgi, a popular Chinese herbal medicine. It is a metabolite of another flavonoid, wogonin [5].
In this study, MTT assays showed that wogonoside did not completely inhibit cell viability; even at 400 µM, viability was inhibited up to 80%. Furthermore, treatment with 50, 100, and 150 µM wogonoside for 96 hours scarcely induced apoptosis in U937 and HL-60 cells, suggesting its antiproliferative effect is primarily due to differentiation and cell cycle arrest rather than direct cytotoxicity [5].
The study demonstrates that wogonoside exerts its antileukemic effects by upregulating PLSCR1 transcription, promoting its nuclear translocation, and enhancing its binding to the IP3R1 promoter, which in turn leads to cell cycle arrest and monocytic differentiation. This mechanism suggests wogonoside may be a potential therapeutic candidate for the treatment of AML [5].

C22H20O11

460.391

460.1

51059-44-0

3084961

Light yellow to yellow solid powder

1.6±0.1 g/cm3

869.0±65.0 °C at 760 mmHg

226-227ºC

304.0±27.8 °C

0.0±0.3 mmHg at 25°C

1.685

1.42

5

11

5

33

763

5

O1[C@]([H])(C(=O)O[H])[C@]([H])([C@@]([H])([C@]([H])([C@]1([H])OC1C([H])=C(C2C(C([H])=C(C3C([H])=C([H])C([H])=C([H])C=3[H])OC=2C=1OC([H])([H])[H])=O)O[H])O[H])O[H])O[H]

LNOHXHDWGCMVCO-NTKSAMNMSA-N

InChI=1S/C22H20O11/c1-30-18-13(32-22-17(27)15(25)16(26)20(33-22)21(28)29)8-11(24)14-10(23)7-12(31-19(14)18)9-5-3-2-4-6-9/h2-8,15-17,20,22,24-27H,1H3,(H,28,29)/t15-,16-,17+,20-,22+/m0/s1

(2S,3S,4S,5R,6S)-3,4,5-trihydroxy-6-(5-hydroxy-8-methoxy-4-oxo-2-phenylchromen-7-yl)oxyoxane-2-carboxylic acid

Oroxindin Wogonoside

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~125 mg/mL (~271.51 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.52 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)