ThePineapple - Journal Club #36: Ischemia | Diabetes | Drug-tests | Senescence

Journal Club #36: Ischemia, Diabetes, Drug-tests, Senescence

Week of 04-03-22

Welcome to journal club. A weekly curation of scientific research papers in the field of cannabis science. Fair warning, these are unedited, freshly destilled scientifc publications. In other words, this post is for the cannabis scientists among you, who are likely going to spent at least one day this week eating free pizza and discussing new publications in your respective biomedical graduate departments. So, if you are a graduate student, feel free to save yourself some time and pick one of these :)

Also, if you feel we missed a particularly juicy paper, then leave a comment with the doi: number or pubmed ID ! We'll update the "journal club" as the week goes on.

Role of PI3K/Akt axis in mitigating hippocampal ischemia-reperfusion injury via CB1 receptor stimulation by paracetamol and FAAH inhibitor in rat.

Abdel Mageed SS, Ammar RM, Nassar NN, Moawad H, Kamel AS.

Neuropharmacology. 2022 Apr 1;207:108935. 

Acetaminophen or paracetamol (PAR), the recommended antipyretic in COVID-19 and clinically used to alleviate stroke-associated hyperthermia interestingly activates cannabinoid receptor (CB1) through its AM404 metabolite, however, to date, no study reports the in vivo activation of PAR/AM404/CB1 axis in stroke. The current study deciphers the neuroprotective effect off PAR in cerebral ischemia/reperfusion (IR) rat model and unmasks its link with AM404/CB1/PI3K/Akt axis. Animals were allocated into 5 groups: (I) sham-operated (SO), (II) IR, (III) IR + PAR (100 mg/kg), (IV) IR + PAR (100 mg/kg) + URB597; anandamide degradation inhibitor (0.3 mg/kg) and (V) IR + PAR (100 mg/kg) + AM4113; CB1 Blocker (5 mg/kg). All drugs were intraperitoneally administered at the inception of the reperfusion period. PAR administration alleviated the cognitive impairment in the Morris Water Maze as well as hippocampal histopathological and immunohistochemical examination of GFAP. The PAR signaling was associated with elevation of anandamide level, CB1 receptor expression and survival proteins as pS473-Akt. P(tyr202/thr204)-ERK1/2 and pS9-GSK3β. Simultaneously, PAR increased hippocampal BDNF and β-arrestin1 levels and decreased glutamate level. PAR restores the deranged redox milieu induced by IR Injury, by reducing lipid peroxides, myeloperoxidase activity and NF-κB and increasing NPSH, total antioxidant capacity, nitric oxide and Nrf2 levels. The pre-administration of AM4113 reversed PAR effects, while URB597 potentiated them.

Significance: PAR poses a significant neuroprotective effect which may be mediated, at least in part, via activation of anandamide/CB1/PI3K/Akt pathway in the IR rat model.

doi: 10.1016/j.neuropharm.2021.108935. Epub 2021 Dec 28. PMID: 34968475.

Spinal cannabinoid CB1 or CB2 receptors activation attenuates mechanical allodynia in streptozotocin-induced diabetic rats.

Gonçalves MR, da Conceição MS, Jesus CHA, Gasparin AT, Rosa ES, da Cunha JM. 

Behav Pharmacol. 2022 Apr 1;33(2&3):158-164. 

Diabetes is a chronic disease associated with a high number of complications such as peripheral neuropathy, which causes sensorial disturbances and may lead to the development of diabetic neuropathic pain (DNP). The current treatment for DNP is just palliative and the drugs may cause severe adverse effects, leading to discontinuation of treatment. Thus, new therapeutic targets need to be urgently investigated. Studies have shown that cannabinoids have promising effects in the treatment of several pathological conditions, including chronic pain. Thus, we aimed to investigate the acute effect of the intrathecal injection of CB1 or CB2 cannabinoid receptor agonists N-(2-chloroethyl)-5Z, 8Z, 11Z, 14Z-eicosatetraenamide (ACEA) or JWH 133, respectively (10, 30 or 100 μg/rat) on the mechanical allodynia associated with experimental diabetes induced by streptozotocin (60 mg/kg; intraperitoneal) in rats. Cannabinoid receptor antagonists CB1 AM251 or CB2 AM630 (1 mg/kg) were given before treatment with respective agonists to confirm the involvement of cannabinoid CB1 or CB2 receptors. Rats with diabetes exhibited a significant reduction on the paw mechanical threshold 2 weeks after diabetes induction, having the maximum effect observed 4 weeks after the streptozotocin injection. This mechanical allodynia was significantly improved by intrathecal treatment with ACEA or JWH 133 (only at the higher dose of 100 μg). Pre-treatment with AM251 or AM630 significantly reverted the anti-allodynic effect of the ACEA or JWH 133, respectively. Considering the clinical challenge that the treatment of DPN represents, this study showed for the first time, that the intrathecal cannabinoid receptors agonists may represent an alternative for the treatment of DNP.

doi: 10.1097/FBP.0000000000000580. PMID: 32804775.

Characterizing and Quantifying Extrahepatic Metabolism of (-)-∆9-Tetrahydrocannabinol (THC) and Its Psychoactive Metabolite, 11-OH-THC.

Kumar AR, Patilea-Vrana GI, Anoshchenko O, Unadkat JD. 

Drug Metab Dispos. 2022 Apr 3:DMD-AR-2022-000868. 

(-)-∆9-Tetrahydrocannabinol (THC) is the psychoactive constituent of cannabis, a drug recreationally consumed by inhalation and orally. Physiologically based pharmacokinetic (PBPK) modeling can be used to predict systemic and tissue exposure to THC and its psychoactive metabolite, 11-OH-THC. To populate a THC/11-OH-THC PBPK model, we previously characterized the depletion clearance of THC (by CYP2C9) and 11-OH-THC (by UGT, CYP3A, and CYP2C9) in adult human liver microsomes. Here we focused on quantifying extrahepatic depletion clearance of THC/11-OH-THC, important after oral (intestine) and inhalational (lung) consumption of THC as well as prenatal THC use (placenta and fetal liver). THC (500 nM) was metabolized in adult human intestinal microsomes (n = 3-5) by CYP2C9 (Vmax: 1.1 {plus minus} 0.38 nmol/min/mg; Km: 70 nM; CLint: 15 {plus minus} 5.4 mL/min/mg and fm: 0.89 {plus minus} 0.31 at concentration << 70 nM) and CYP3A (CLint: 2.0 {plus minus} 0.86 mL/min/mg; fm: 0.11 {plus minus} 0.050). 11-OH-THC (50 nM) was metabolized by CYP3A (CLint: 0.26 {plus minus} 0.058 mL/min/mg; fm: 0.51 {plus minus} 0.11) and UGT2B7 (CLint: 0.13 {plus minus} 0.027 mL/min/mg; fm: 0.25 {plus minus} 0.053). THC at 500 nM (CLint: 4.7 {plus minus} 0.22 mL/min/mg) and 11-OH-THC at 50 nM (CLint: 2.4 {plus minus} 0.13 mL/min/mg) were predominately (fm: 0.99 and 0.80, respectively) metabolized by CYP3A in human fetal liver microsomes (n = 3). However, we did not observe significant depletion of THC/11-OH-THC in adult lung, 1st, 2nd trimester or term placentae microsomes. Using PBPK M&S, these data could be used in the future to predict systemic and tissue THC/11-OH-THC exposure in healthy and special populations. 

Significance : This is the first characterization and quantification of THC and 11-OH-THC depletion clearance by CYP and UGT enzymes in extrahepatic human tissues: intestine, fetal liver, lung, and placenta. These data can be used to predict, through PBPK M&S, systemic and tissue THC/11-OH-THC exposure, after inhalational and oral THC use, in both healthy and special populations (e.g. pregnant women).

doi: 10.1124/dmd.122.000868. Epub ahead of print. PMID: 35370140.

CB2R Attenuates Intervertebral Disc Degeneration by Delaying Nucleus Pulposus Cell Senescence through AMPK/GSK3β Pathway.

Du J, Xu M, Kong F, Zhu P, Mao Y, Liu Y, Zhou H, Dong Z, Yu Z, Du T, Gu Y, Wu X, Geng D, Mao H. 

Aging Dis. 2022 Apr 1;13(2):552-567.

Nucleus pulposus (NP) cell (NPC) senescence is one of the main causes of intervertebral disc degeneration (IVDD). However, the underlying mechanism of NPC senescence is still unclear. The cannabinoid type 2 receptor (CB2R) is a member of the cannabinoid system and plays an important role in antioxidative stress, anti-inflammatory and antisenescence activities. In this study, we used a hydrogen peroxide (H2O2)-induced NPC senescence model and a rat acupuncture IVDD model to explore the role of CB2R in IVDD in vitro and in vivo. First, we confirmed that the expression of p16INK4a in the NP tissues of IVDD patients and rat acupuncture IVDD models obviously increased accompanied by a decrease in CB2R expression. Subsequently, we found that activation of CB2R significantly reduced the number of SA-β-gal positive cells and suppressed the expression of p16INK4a and senescence-related secretory phenotypes [SASP, including matrix metalloproteinase 9 and 13 (MMP9, MMP13) and high mobility group protein b1 (HMGB1)]. In addition, activation of CB2R promoted the expression of collagen type II (Col-2) and SRY-Box transcription factor 9 (SOX9), inhibit the expression of collagen type X (Col-X), and restore the balance of extracellular matrix (ECM) metabolism. In addition, the AMPK/GSK3β pathway was shown to play an important role in CB2R regulation of NPC senescence. Inhibition of AMPK expression reversed the effect of JWH015 (a CB2R agonist). Finally, we further demonstrated that in the rat IVDD model, the AMPK/GSK3β pathway was involved in the regulation of CB2R on NPC senescence. In conclusion, our experimental results prove that CB2R plays an important role in NPC senescence. Activation of CB2R can delay NPC senescence, restore the balance of ECM metabolism, and attenuate IVDD.

doi: 10.14336/AD.2021.1025. PMID: 35371598; PMCID: PMC8947828.