2022.07.20

[기억할 것]

• 기계 욕심을 부리지 말자. 협업으로 극복하자. learned helplessness model
• 나에게 친절하자. 운동, 휴식, 가꾸기 등
• 잘하려고 경직되지 말자. 
• 실험 디자인
• 기록을 해놓자. 중요한 것과 중요하지 않은 것이 구별된다. 
• 기업: 연구지원기업, 맞춤형 향기업

[마무리할 것]

• SPS
• KNU
• prolonged SDS
• Dr. oh. 해부과정. 전문가초빙. cortex, amygdala, hippocampus

[Article]

Beneficial effects of prolonged 2-phenylethyl alcohol inhalation on chronic distress-induced anxio-depressive-like phenotype in female mice. Biomedicine & Pharmacotherapy. 2022

#Animal model

• the beneficial effect of PEA is restricted to the existence of pathological traits 
• It has been shown that naive male C57BL/6 mice are unresponsive to chronic fluoxetine, a selective serotonin re-uptake inhibitor used in the treatment of depression, but the CORT-induced activation of stress circuitry is sufficient to reveal its therapeutic action in this typically unresponsive strain

#Duration

• most animal studies focusing on rose odor essential oil effects, or its active constituents, have been carried out in naive animals during or immediately after acute odor inhalation.
• in clinical trials aromatherapy is rather prescribed over a long period of time, i.e. in a chronically administrated manner. 

#Administration

• inhalation allows avoiding the additional stress associated with intraperitoneal injection, the delivery method used in the majority of preclinical studies

#Female mice

• data concerning the behavioral and neural changes occurring in female mice are scarce.
• the present study did not reveal neither a strong anxiogenic- nor a marked depressant-like effect of chronic CORT administration in female C57BL/6JRJ mice, when they were evaluated in the OF and FST, respectively.
• dysfunction on motor performance

#Novelty-suppressed feeding test

• Besides being sensitive to anxiolytic drugs, the NSF task is also known to respond to chronic, but not acute, antidepressant treatments.
• Thus, it is considered to be a reliable behavioral paradigm with predictive validity for investigating the therapeutic action of antidepressants that requires several weeks to emerge 

#cFos expression in the olfactory bulb (OB).

• these behavioral and functional alterations are further associated with a pronounced reduction in hippocampal and OB neurogenesis, but reversed by prolonged antidepressant treatment with fluoxetine.
• Similarly, in the present study, we revealed that both anxio-depressive-like phenotype and decreased OB activity were reversed by chronic PEA inhalation, suggesting that the therapeutic action of PEA is mainly exerted via the olfactory pathway.
• PEA is known to possess no intranasal trigeminal properties and to activate selectively the olfactory system 

#appetite #food intake

• As part of the limbic corticostriatal circuitry, it is also known to be implicated in motivational-based appetitive behaviors [21], [88]. 
• olfactory processing involves a large number of strongly interconnected brain regions that incorporate complex overlapping networks [86], thus creating functional interactions between neural circuits controlling emotional and feeding processes [98], [99], [100], [101].

#mechanism #reserpine

• rose oil induces an anxiolytic-like effect similar to benzodiazepines [34], [35] but its mechanism of action seems to be independent of the recruitment of functional GABA-A receptors [34]. 
• It has been also suggested that the antidepressant-like activity of rose oil may result from a modulation of monoaminergic neurotransmission mediated through a pre-synaptic mechanism since its effect in the FST was inhibited by an injection of reserpine
• The antidepressant activity of fluoxetine in the NSF task in CORT-treated mice involves neurogenesis-dependent mechanisms

#Brain 

• The amygdala and the entorhinal cortex receive direct bulbar projections, thus participating actively in odor information processing [86]. Moreover, it has been shown that the amygdala plays a key role in encoding the emotional salience of the olfactory stimulus.
• olfactory processing involves a large number of strongly interconnected brain regions that incorporate complex overlapping networks [86], thus creating functional interactions between neural circuits controlling emotional and feeding processes [98], [99], [100], [101]. Therefore, it cannot be excluded that the OB neural activity changes could originate from a differential action of centrifugal systems

#Graph notation. (Bar chart) Fig. 2A

Yueju Pill Rapidly Induces Antidepressant-Like Effects and Acutely Enhances BDNF Expression in Mouse Brain. Evidence-Based Complementary and Alternative Medicine. 2013

#Rapid #Quick #long-lasting #sustained

• conventional antidepressants, this effect does not last long, and it takes a long period time of repeated administrations before the antidepressant effects manifest in depressive patients or animals 
• Conventional monoamine-based antidepressants also increase BDNF, but only after chronic drug administration
• Previously, it has been reported that ethanol, but not aqueous, extract of Yueju has an antidepressant effect after a week-long administration [14]. Here, we provide further evidence that a single dose of the ethanol extract of Yueju has a quick and lasting antidepressant effect.

#Learned Helplessness #rapid #Fig. 2

• For the induction of helplessness, mice received 120 inescapable shocks (18–44 s, average 30 s; 0.45 mA for 15 s) once daily for 2 consecutive training days. For screening of helpless mice, animals were subjected to 30 avoidance trials (18–44 s, average 30 s; 0.45 mA for 3 s). 
• we found improvement of active avoidance escape responses and reducing freezing time 
• Mice that developed helplessness (>10 escape failures) were treated with either Yueju or vehicle and were tested for helplessness 24 hours after the treatment.
• In LH paradigm, only after several days of repeated administration of conventional monoaminergic antidepressants, mice start to show response to the treatment

#Sustained #Fig. 3

• At 24 hours after administration
• At 30 minutes after drug administration in independent group
• By 48 hours after drug administration
• open field behavior did not differ (30 min, 24 h) → What is a good behavioral assay to test the quick rapid acting antidepressant effect?
• only higher doses (30 mg/kg) have been reported to induce lasting antidepressant effect using tail suspension test paradigm

#Mechanism underlying rapid effect #BDNF #Fig. 4

• The rapid and enduring antidepressant effect of ketamine is also dependent on BDNF.
• Previous report suggests that ketamine deactivates eukaryotic elongation factor 2 (eEF2) kinase, leading to reduced eEF2 phosphorylation and desuppression of translation of BDNF.
• Conventional monoamine-based antidepressants also increase BDNF, but only after chronic drug administration.
• we examined the expression of BDNF in the hippocampus at 30 minutes, 24 hours, and 48 hours after drug administrations. 
• We identified the fast induction of BDNF expression as the key molecule by which Yueju exerts rapid antidepressant effects, but the upstream signaling as well as the contributions of other forms of neuroplasticity remain unknown. For example, mTOR signaling is also implicated in rapid and lasting antidepressant response.
• Phosphorylation of eEF2 suppresses translation of proteins including BDNF. We found that Yueju rapidly downregulated the phosphorylation of eEF2. This dephosphorylation likely augments BDNF expression and thus exerts antidepressant effects 
• increase of BDNF expression is likely crucial for onset of antidepressant effects but may not be required for the sustaining antidepressant effects.
• In contrast to the increase in protein expression, the BDNF gene expression did not alter.
• Taken together, it is likely that the increase in BDNF protein expression resulted from posttranscriptional regulation.

#Mismatch between molecular level and behavior

• There was no change in open field test or cage activities in Yueju-treated mice at 24 hours with reduced hippocampal BDNF, which returned to normal level at 48 hours. It remained to know the other potential behavioral consequences from the temporary decrease of BDNF expression by Yueju.

#Strain

• The possibility of a longer antidepressant effect of Yueju in other mouse strains cannot be ruled out.

Hypidone Hydrochloride (YL-0919) Produces a Fast-Onset Reversal of the Behavioral and Synaptic Deficits Caused by Chronic Stress Exposure. Front Cell Neurosci. 2018

#Two models. #normal & CUS #fig 1. 

#sensitive behavioral test #TST, FST #SPT #(OPT)

• TST, FST → Good ways to measure the rapid-acting antidepressant effect? YL-0919 Rapidly Reversed Acute Behavioral Deficits in Mice. (Fig. 1)
• The SPT is a sensitive and reliable method to study the time course of antidepressant efficacy
• In contrast to acute treatment, chronic treatment with antidepressants reduces hyponeophagia.

#standard for fast/slow acting

• YL-0919 Rapidly Reversed the CUS-Induced Behavioral Deficits in Rats (Fig. 2)
• Treatment with YL-0919 (2.5 mg/kg, i.g.), but not with fluoxetine (Flx; 10 mg/kg, i.g.), caused a fast improvement in the SPT scores. In CUS-exposed rats, YL-0919 treatment for 5 days decreased the immobility time in a forced swimming test (FST), and a 10-day treatment decreased the latency to feed in a Novelty-Suppressed Feeding Test (NSFT)
• We found that YL-0919 was able to increase the sucrose preference rate after only 3 days of treatment, while the first-line SSRI Flx did not show this rapid action. Even 10 days of treatment with Flx could not increase the sucrose preference rate significantly. This is in accordance with studies reporting a time lag of 2–4 weeks before currently available medications exert an antidepressant effect (Penn and Tracy, 2012).

#sustained effects?? 

• YL-0919 showed a powerful effect in the SPT from the third day onward after drug treatment, which implies that it has sustained effects 
• Rats were exposed to CUS for 35 days and administered YL-0919 (2.5 mg/kg, i.g.) or Flx (10 mg/kg, i.g.) from day 21 onwards and lasting for 14 days

#Rapamycin #Fig. 3

• These effects of YL-0919 were completely blocked by rapamycin treatment.
• Rapamycin given for 35 days, in the absence of YL-0919 and in stressed rats, had no effect in either test.

#Western Blotting #appropriate timepoint?

• Hippocampi of rats from different groups were dissected on the 35th CUS day.

#5-HT

• indirect reduction in serotonergic transmission via the activation of somatodendritic 5-HT1A receptors in the raphe nuclei, with resulting reduction of 5-HT release, plays an important role in the delayed onset of therapeutic SSRI actions. It has been suggested that 5-HT partial agonist and reuptake inhibitor may theoretically speed up the onset of antidepressant-like activity

Vilazodone for the Treatment of Major Depressive Disorder: Focusing on Its Clinical Studies and Mechanism of Action. Psychiatry Investig. 2015

Opposite role of pre-and postsynpatic 5HT1A receptors in depression. 1) Activation of pre-synaptic 5HT1A receptors, autoreceptors, decrease the firing and secretion of 5-HT, where as 2) activation of post-synaptic 5HT1A receptors increase firing and secretion of 5HT, 3) Acute SSRI administration results in net increase of extracellular 5HT, 4) which is immediately counteracted by neuronal negative feedback mechanisms mediated by 5-HT1A autoreceptors causing therapeutic lag of SSRI. 5HT: serotonin, SERT: serotonin transporter. Vilazodone for the Treatment of Major Depressive Disorder: Focusing on Its Clinical Studies and Mechanism of Action. 2015

Vilazodone's mechanism of action. By acting only as a partial agonist at 5-HT1A autoreceptors, vilazodone may more rapidly desensitize 5-HT1A autoreceptors without causing excess activation of 5-HT1A autoreceptor-mediated serotonin inhibition. In 5-HT1A postsynaptic receptors, vilazodone will enhance 5-HT. Synergistic with its SSRI properties, vilazodone would yield even more serotonin release. 5HT: serotonin, SERT: serotonin transporter, SSRI: selective serotonin reuptake inhibitor. Vilazodone for the Treatment of Major Depressive Disorder: Focusing on Its Clinical Studies and Mechanism of Action. 2015

Mechanism of action of antidepressants. Psychopharmacol Bull. 2002

• It is hoped that the development of drugs with multiple sites of action will translate into improved clinical efficacy in the treatment of depression.

Effect of the fragrance inhalation of essential oil from Asarum heterotropoides on depression-like behaviors in mice. BMC Complement Altern Med. 2015

#Remove n-hexane

• The pulverized Asarum heterotropoides were used to extract essential oil at the room temperature with n-hexane (5 L x 2) for 24 h. The extracts (EOAH) were filtered and evaporated in the vacuum at 40°C to remove n-hexane.

#Inhalation method

• Male ICR mice received fragrance inhalation of EOAH (0.25, 0.5, 1.0, and 2.0 g) for 3 h in the special cage capped with a filter paper before start of the forced swimming test (FST) and tail suspension test (TST). 
• To induce saturation of the fragrance by EOAH in the transparent special cage (W 26 X L 22 X H 20 cm), EOAH put 30 min before the individual inhalation of EOAH for 3 h in the special cage under standardized condition (room temperature: 23 ± 2°C, relative humidity: 50 ± 10%). 

#fluoxetine #15 mg/kg #(i.p.)

• fluoxetine markedly reduced immobility time at both the FST and TST compared to saline group
• fluoxetine as positive control was administered at a dose of 15 mg/kg (i.p.) 30 min before start of behavioral testing.

#Immunohistochemistry #time

• At the end of the FST, mice were sacrificed and brains were prepared to measure the expressions of 5-HT, TH, and CRF, respectively.

#we will...

• Rapid-acting (immediately after acute odor inhalation. in Naive animal model) 
• SDE inhalation (or i.n) vs Aqueous extract p.o vs fluoxetine i.p
• EPM, OFT, TST, FST, Y-maze, Social interaction
• Mechanism 
1. A much larger decrease in the mean number of 5-HT-immunoreactive neurons in the dorsal raphe nucleus was produced in mice exposed to the FST compared with normal mice. Effect of the fragrance inhalation of essential oil from Asarum heterotropoides on depression-like behaviors in mice. 2015
2. Olfactory Input *anosmic mice Linalool Odor-Induced Anxiolytic Effects in Mice. 2015
3. inhibitor eg. flumazenil(GABA) WAY100635(5-HT) Linalool Odor-Induced Anxiolytic Effects in Mice. 2015
4. in vitro study? 
5. To understand the molecular mechanism underlying rapid antidepressant effect of Yueju, we examined the expression of BDNF in the hippocampus at 30 minutes, 24 hours, and 48 hours after drug administrations. Yueju Pill Rapidly Induces Antidepressant-Like Effects and Acutely Enhances BDNF Expression in Mouse Brain. 2013
• ref. 
1. Linalool Odor-Induced Anxiolytic Effects in Mice. 2018
2. Anxiolytic Effect of Two Tobacco Essential Oils (Nicotiana tabacum Linn.) on Mice. 2021
3. Behavioural effects of inhalation exposure to dizocilpine (MK-801) in mice. 2019
4. Anti-depressive-like effect of 2-phenylethanol inhalation in mice. 2019
5. Effect of the fragrance inhalation of essential oil from Asarum heterotropoides on depression-like behaviors in mice. 2015
6. Yueju Pill Rapidly Induces Antidepressant-Like Effects and Acutely Enhances BDNF Expression in Mouse Brain. 2013

• Prolonged effect
• LPS model(?) CORT model(?) 
• NSF task is also known to respond to chronic, but not acute, antidepressant treatments. Beneficial effects of prolonged 2-phenylethyl alcohol inhalation on chronic distress-induced anxio-depressive-like phenotype in female mice. 2022
• Dose effect only higher doses (30 mg/kg) have been reported to induce lasting antidepressant effect using tail suspension test paradigm. Yueju Pill Rapidly Induces Antidepressant-Like Effects and Acutely Enhances BDNF Expression in Mouse Brain. 2013

Sleep-enhancing Effects of Phytoncide Via Behavioral, Electrophysiological, and Molecular Modeling Approaches

• 서수현(?) 

Inhalation chamber

1. Inhalation treatment of lung cancer: the influence of composition, size and shape of nanocarriers on their lung accumulation and retention
2. Ethnopharmacological Evaluation of Breu Essential Oils from Protium Species Administered by Inhalation