Antiallodynic and antihyperalgesic effects of LMH-2, a new sigma-1 receptor antagonist, on the neuropathic pain of diabetic mice

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Mechanical allodynia was obtained through the up-and-down method using a sequence of von Frey filaments with different forces and logarithmic increments (0.008 to 10 g). Each von Frey filament (starting with 0.008 g) was placed perpendicular to the midplantar surface of the right hind paw with enough force to cause a slight curvature of the filament against the paw. It was maintained from 5 to 8 sec. In the case of a positive response (paw withdrawal), the next filament with a smaller force was applied; in the case of a negative response (no paw withdrawal response), the next filament with greater force was applied This was repeated until a total of 5 applications [21]. The 50% of the withdrawal threshold was expressed in grams (g), and it was calculated according to the following formula 50% of the withdrawal threshold (g) = (10^[Xf+Kδ]/10,000) Where: Xf: Value in grams of the last Von Frey filament used in logarithmic units K: Correction factor indicated in the table published by Dixon for the pattern of positive and/or negative responses obtained in the test δ: Mean difference between the response pattern obtained in logarithmic units To determine hyperalgesia in the hyperglycemic mice, the formalin test with a brief modification was used one day after the mechanical allodynia determination. For this, mice received 20 µL of 0.5% formalin solution subcutaneously (s.c.) into the right hind paw, and the number of flinches of the injected paw was quantified every 5 min for 30 min.

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