The neuroprotective effect of Incensole acetate against Aβ-induced neurotoxicity is associated with activation of the Erk1/2 and Nrf2 signaling pathways, leading to induction of heme oxygenase-1 (HO-1) expression. [1]

Aβ25-35-induced cell death can be markedly enhanced by acetic acid incense (100 μM; pretreatment for 4 hours). The viability of Aβ25-35 cells dropped by 48-52%, but it could be recovered to 85-89% with IA pretreatment. 1.]. In hOBNSC, incensoleacetate (5-100μM) pretreatment reverses the increase in Bax, caspase 8, cyto c, and mRNA and protein levels induced by Aβ 25-35, as well as the decrease in Bcl2 mRNA and protein levels [1].

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Incensole acetate (100 μM, 4h pretreatment) stimulated proliferation of hOBNSCs, as revealed by a robust increase in expression of proliferation-related genes Nestin (Nes) and Sox2, and increased the number and size of neurospheres compared to Aβ25-35-treated cells. [1]
Incensole acetate (100 μM, 4h pretreatment) induced differentiation of neuronal progenitors in hOBNSCs, marked by increased expression of the immature neuronal marker Map2, while notably downregulating the astrocyte marker Gfap to levels similar to control cells. [1]
Incensole acetate (100 μM, 4h pretreatment) significantly ameliorated Aβ25-35 (5 or 10 μM, 24h)-induced cell viability loss, restoring viability from 48-52% decrease to 85-89% as measured by MTT assay. [1]
Incensole acetate (100 μM, 4h pretreatment) reduced Aβ25-35-triggered apoptosis, as evidenced by decreased caspase 3 enzymatic activity and protein expression level, downregulated mRNA and protein expression of pro-apoptotic markers Bax, caspase 8, and cytochrome c (cyto c), and upregulated expression of the anti-apoptotic marker Bcl2. [1]
Incensole acetate (100 μM, 4h pretreatment) restored the impaired mitochondrial membrane potential (MMP) induced by Aβ25-35 (revealed by elevated rhodamine 123 fluorescence) and lowered the elevated intracellular Ca2+ level (measured by Fura-2/AM imaging). [1]
Incensole acetate (100 μM, 4h pretreatment) attenuated Aβ25-35-induced oxidative stress by scavenging intracellular reactive oxygen species (ROS), reducing the elevated malondialdehyde (MDA) level, and increasing the activities of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX). [1]
Incensole acetate (100 μM, 4h pretreatment) ameliorated Aβ25-35-induced inflammation, as marked by downregulated mRNA expression of pro-inflammatory genes IL1b, TNFα, NFkB, and Cox2. [1]
Incensole acetate (100 μM, 4h pretreatment) significantly upregulated mRNA and protein expression of Erk1/2, Nrf2, and the downstream antioxidant enzyme heme oxygenase-1 (HO-1). The neuroprotective effect of Incensole acetate against Aβ25-35-induced cytotoxicity was reversed by the HO-1 inhibitor zinc protoporphyrin (ZnPP), as measured by MTT assay. [1]

Cell viability assay [1]
Cell Types: human olfactory bulb neural stem cells
Tested Concentrations: 100 μM
Incubation Duration: 4 hrs (hours) of pretreatment
Experimental Results: Aβ 25-35 induced an increase in hOBNSC cell viability.

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Human olfactory bulb neural stem cells (hOBNSCs) were obtained from adult patients and cultured as neurospheres in proliferation medium (DF12 containing EGF, bFGF, and LIF) or differentiation medium (DF12 without growth factors). For experiments, cells were pretreated with Incensole acetate (100 μM) for 4 hours, followed by co-incubation with Aβ25-35 (5 or 10 μM) for 24 hours. [1]
Cell viability was assessed using the MTT assay. MTT stock solution was added to cells in 96-well plates, and the absorbance of extracted formazan in DMSO was recorded at 570 nm. [1]
Intracellular reactive oxygen species (ROS) was measured using the fluorescent probe 2,7-dichlorofluorescein diacetate. Mitochondrial membrane potential (MMP) was detected using rhodamine 123 (Rh123) fluorescence. Intracytoplasmic Ca2+ level was measured using imaging of the fluorescent probe Fura-2/AM. Levels of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) were measured using commercial kits. [1]
Caspase 3 activity was measured by incubating cell lysates with a specific fluorogenic substrate (7-amino-4-methylcoumarin N-acetyl-L-aspartyl-L-glutamyl-L-valyl-L-aspartic acid amide). Cleavage was detected using a fluorescence microtitre plate reader at excitation/emission wavelengths of 360/460 nm. [1]
Relative gene expression was detected by real-time PCR. Total RNA was isolated, reverse transcribed to cDNA, and qPCR was performed using SYBR Green Master Mix with specific primers for Nes, Sox2, Map2, Gfap, Bax, caspase 8, cyto c, Bcl2, TNFα, IL1β, Cox2, NFkB, HO-1, Nrf2, Erk2, and β-actin. The 2-ΔΔCt method was used for analysis. [1]
Western blotting was performed using primary antibodies against Bax, caspase 3, cyto c, caspase 8, Bcl2, HO-1, phosphor-ERK1/2, Nrf2, and B actin. Secondary horseradish peroxidase-conjugated anti-rabbit IgG antibody detection was done with enhanced chemiluminescence reagents. Band density was measured using Image J software. [1]
To assess the role of HO-1, cells were pre-incubated with the HO-1 inhibitor zinc protoporphyrin (ZnPP) for 1 hour, then with Incensole acetate (100 μM) for 4 hours, and finally with Aβ25-35 for 24 hours, after which cell viability was determined by MTT assay. [1]

[1]. Incensole acetate prevents beta-amyloid-induced neurotoxicity in human olfactory bulb neural stem cells. Biomed Pharmacother. 2018 Sep;105:813-823.

Incensole acetate is a major constituent of Boswellia carterii resin, a herb used for centuries in traditional medicine. It has been shown to have neuroprotective effects against hippocampal neurodegeneration and can improve cognitive ability in traumatic and ischemic head injuries. [1]
The study demonstrates that Incensole acetate can induce proliferation and differentiation of neuronal progenitors in hOBNSCs and protect these cells from Aβ25-35-induced oxidative cell death, at least in part, through induction of HO-1 expression via activation of Erk1/2 and Nrf2 signal pathways. [1]
Incensole acetate may be applicable as a potential preventive agent for Alzheimer's disease (AD) by its effect on hOBNSCs and could also be used as an adjuvant to hOBNSCs in cellular therapy of neurodegenerative diseases. [1]

C22H36O3

348.5194

348.266

C, 75.82; H, 10.41; O, 13.77

34701-53-6

53386731

Colorless to light yellow liquid

1.0±0.1 g/cm3

420.1±45.0 °C at 760 mmHg

178.6±23.3 °C

0.0±1.0 mmHg at 25°C

1.499

6.83

0

3

3

25

546

3

O1C2(C([H])([H])[H])C([H])(C([H])([H])C([H])([H])C(C([H])([H])[H])=C([H])C([H])([H])C([H])([H])C(C([H])([H])[H])=C([H])C([H])([H])C1(C([H])(C([H])([H])[H])C([H])([H])[H])C([H])([H])C2([H])[H])OC(C([H])([H])[H])=O

HVBACKJYWZTKCA-XSLBTUIJSA-N

InChI=1S/C22H36O3/c1-16(2)22-13-12-18(4)9-7-8-17(3)10-11-20(24-19(5)23)21(6,25-22)14-15-22/h8,12,16,20H,7,9-11,13-15H2,1-6H3/b17-8+,18-12+/t20-,21+,22+/m0/s1

[(1R,2S,5E,9E,12S)-1,5,9-trimethyl-12-propan-2-yl-15-oxabicyclo[10.2.1]pentadeca-5,9-dien-2-yl] acetate

Incensol Acetate; Incensole Acetate

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 : ~100 mg/mL (~286.93 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.17 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 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 25.0 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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Solubility in Formulation 2: 2.5 mg/mL (7.17 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (7.17 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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