Effect of ABT-639 on Cav3.2 channel activity and its analgesic actions in mouse models of inflammatory and neuropathic pain.

Cav3.2 T-type calcium channels are important targets for pain relief in rodent models of inflammatory and neuropathic pain. Even though many T-type channel blockers have been tested in mice, only one molecule, ABT-639, has been tested in phase II clinical studies and did not produce analgesic effects over placebo. Here we examined the effects of ABT-639 on Cav3.2 channel activity in tsA-201 cells and dorsal root ganglion (DRG) neurons, in comparison with another established Cav3.2 inhibitor Z944. These experiments revealed that Z944 mediated ∼100-fold more potent inhibition of Cav3.2 currents than ABT-639, with the latter blocking channel activity by less than 15 percent when applied at a concentration of 30 µM. A slight increase in ABT-639 potency was observed at more depolarized holding potentials, suggesting that this compound may act preferentially on inactivated channels. We tested the effects of both compounds in the Complete Freund's Adjuvant (CFA) model of chronic inflammatory pain, and in partial sciatic nerve injury model of neuropathic pain in mice. In the neuropathic pain model, both Z944 and ABT-639 reversed mechanical hypersensitivity to similar degrees when delivered systemically, but remarkably, when delivered intrathecally, only Z944 was effective. In the CFA model, both compounds reversed thermal hyperalgesia upon systemic delivery, but only Z944 mediated pain relief upon intrathecal delivery, indicating that ABT-639 acts primarily at peripheral sites. ABT-639 lost its analgesic effects in CFA treated Cav3.2 null mice, indicating that these channels are essential for ABT-639-mediated pain relief despite its poor inhibition of Cav3.2 currents.

Analgesia by intrathecal delta-9-tetrahydrocannabinol is dependent on Cav3.2 calcium channels.

Delta-9-tetrahydrocannabinol (Δ9-THC) is known to produce systemic analgesia that involves CB1 and CB2 cannabinoid receptors. However, there is compelling evidence that Δ9-THC can potently inhibit Cav3.2T-type calcium channels which are highly expressed in dorsal root ganglion neurons and in the dorsal horn of the spinal cord. Here, we investigated whether spinal analgesia produced by Δ9-THC involves Cav3.2 channels vis a vis cannabinoid receptors. We show that spinally delivered Δ9-THC produced dose-dependent and long-lasting mechanical anti-hyperalgesia in neuropathic mice, and showed potent analgesic effects in models of inflammatory pain induced by formalin or Complete Freund's Adjuvant (CFA) injection into the hind paw, with the latter showing no overt sex differences. The Δ9-THC mediated reversal of thermal hyperalgesia in the CFA model was abolished in Cav3.2 null mice, but was unaltered in CB1 and CB2 null animals. Hence, the analgesic effects of spinally delivered Δ9-THC are due to an action on T-type calcium channels, rather than activation of spinal cannabinoid receptors.

Benefits of electroacupuncture and a swimming association when compared with isolated protocols in an osteoarthritis model.

Background and aim: Osteoarthritis (OA) is characterized by pain and inflammation. Electroacupuncture (EA) and swimming (SW) are non-pharmacological interventions recommended for treating OA. The study evaluated the benefits of electroacupuncture (EA) and swimming (SW) association when compared with isolated protocols in an OA rodent model. Experimental. Procedures: An ankle monoarthritis model was induced in rats by applying Complete Freund's Adjuvant (CFA). After seven days of induced OA, the groups were submitted to EA (ST36 and the GB 30 Acupoint), SW, or the EA + SW protocol. The nociceptive behavior was measured by the Von Frey test, the Cold Stimulation test, and the Paw Flick Immersion test. Inflammatory activity was evaluated by measuring TNF levels, myeloperoxidase, NAGase, immunological parameters and the histology from the subcutaneous tissue. Results: Compared to CFA group, EA decreased the nociceptive scores in the cold stimulation test (p < 0.05), and it also increased the latency time in thermal cold (p < 0.01) and heat hyperalgesia (p < 0.001). Also, EA reduced NAGase (p < 0.01). SW reduced the edema (p < 0.05) and did not increase the inflammatory infiltrates or congestion, neither in the histological measurements nor by analyzing the levels of TNF. The association of EA + SW decreased the neutrophils and the monocytes, MPO (p < 0.05), and the glutamate levels in the cerebrospinal fluid (CSF, p < 0.001). Conclusion: There were statistical differences between combination therapy and monotherapy as seen by the inflammatory parameters, which could be associate to the delay of the chronification osteoarthritis retardation. However, EA + SW did not show benefits when compared to isolated protocols in nociceptive behavior.

Evaluation of the efficacy and toxicity of oral and topical pumpkin oil on the hair growth of mice.

This study aimed to evaluate the efficacy and safety of a topical and oral administration of pumpkin seed oil (PSO) on the hair growth of BALB/c male mice. The animals had their dorsal area shaved (2 ×2 cm) and they were divided into 6 experimental groups. They received orally saline (OS), finasteride (F), or PSO (OP) for 14 days; or topically saline (TS), minoxidil (M), or PSO (TP) for 7 days. The euthanasia of all of the mice occurred on the 22nd day, and the histological slides from the skin area were analyzed. Lipoperoxidation in the liver was assessed through the TBARS method and was also evaluated by the antioxidant enzymes (SOD and CAT). The comet assay and the micronucleus tests were performed for genotoxic/mutagenic safety analyses. A significant increase in the number of hair follicles in the TP group was seen (8.8 ± 0.8) but it was disorganized, with loose dermal collagen. Finasteride presented a significant increase in the levels of the TBARS, SOD, and CAT in the liver, and M increased the DNA damage in the blood and the liver tissues. PSO did not induce any significant changes. In addition, PSO did not induce genotoxic or mutagenic effects. In conclusion, the oral PSO for 14 days acted in the proliferation of the hair follicles, without toxicity signals in the liver. DATA AVAILABILITY: The authors confirm that all of the relevant data is included in the article and/or in the supplementary information file.

Evaluation of lipolysis and toxicological parameters of low-level laser therapy at different wavelengths and doses in the abdominal subcutaneous tissue.

Investigate the effects of low-level lasers therapy (LLLT) aiming abdominal lipolysis. Female Wistar rats received applications of LLLT directly in the abdominal skin twice a week (5 weeks). Except the control group (n = 5), animals received treatments with red wavelength 660 nm being (I) R3.3 group (n = 5): 3.3 J/cm2, and (II) R5 group (n = 5): 5 J/cm2, or infrared wavelength 808 nm being (III) IR3.3 group (n = 5): 3.3 J/cm2, and (IV) IR5 group (n = 5): 5 J/cm2. Abdominal subcutaneous and liver tissues were evaluated histologically. Levels of thiobarbituric acid reactive substances (TBARS) and catalase (CAT) activity were analyzed in liver tissue. In the peripheral blood aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglycerides, and total cholesterol were investigated. Micronucleus assay was performed in the bone marrow. Except for the IR3.3 group, all treated groups reduced the body weight (p < 0.001). The R5 group reduced the abdominal subcutaneous tissue weight and thickness (p < 0.05), even though all treated groups reduced the number of adipocytes and its size (p < 0.001). No histological changes in the liver. There were no alterations in the triglycerides and LDL levels. The IR5 group increased the total cholesterol levels and decreased the HDL, ALT (both p < 0.05), and AST levels (p < 0.001). The group IR3.3 showed higher levels of ALP (p < 0.01). The R3.3 group increased the TBARS and CAT activity (p < 0.05). No mutagenic effects were found. The red laser treatment at 5 J/cm2 led to lipolysis and did not alter the liver's parameters.