Clemastine Fumarate

Cell lines

Preparation Method

RBL-2H3 cells were seeded into 96-well flat bottom culture plates at a density of 1×10⁴ cells/well and incubated for 24h. Before stimulation with C48/80 (10μg/ml, used to promote MC degranulation) for 30min, cells were treated with Clemastine Fumarate (1ng/ml - 10μg/ml). Tryptase is a specific mediator of MC degranulation. The released tryptase was obtained by centrifuging the cells at 2000rpm for 10min at 4°C. Supernatants (100μl) were added to 100μl of 0.8mmol/l α-N-benzoyl-L-arginine-p-nitroanilide in Tris buffer and incubated at 37°C for 72h. C48/80 induces lethal degranulation of MCs. Therefore, Cell Counting Kit-8 (CCK-8) was used as one of the indicators for measuring MC degranulation.

Reaction Conditions

Applications

Animal models

Preparation Method

After the abdominal hair was removed, a 1.5-cm incision was made along the midline of the abdomen in adult male Sprague Dawley rats to fully expose the cecum. Subsequently, the cecum was ligated and perforated using a sterile 22-gauge needle. Finally, the abdominal musculature and skin were sutured. The sham group only underwent the previously described laparotomy and cecum exposure.
Three different doses of Clemastine Fumarate (10mg/kg, 30mg/kg and 50mg/kg) were dissolved in saline solution and administered via intraperitoneal injection 30min after CLP surgery. 15mg/kg 3-methyladenine (3-MA) was administered via intraperitoneal injection to rats in the CLP + Clemastine Fumarate group immediately after CLP surgery. Equal volumes of normal saline (NS) were used as controls.
After acclimating to the housing environment for one week, the rats were randomly assigned to groups using a random number table as follows: 1) To evaluate whether Clemastine Fumarate improves survival rates and decreases serum cTnI levels in septic rats and to observe the effects of different doses of Clemastine Fumarate, the rats were randomly divided into the following groups: (1) Sham + NS; (2) CLP + NS; (3) CLP + Clemastine Fumarate-10; (4) CLP + Clemastine Fumarate-30; and (5) CLP + Clemastine Fumarate-50; 2) After determining the optimal dose, to assess the effects of Clemastine Fumarate treatment on echocardiography, hematoxylin-eosin (H&E) staining, apoptosis, autophagy, and mitochondrial damage, the rats were randomly divided into another three groups: (1) Sham + NS; (2) CLP + NS; and (3) CLP + Clemastine Fumarate group; 3) To evaluate whether Clemastine Fumarate functions in myocardial apoptosis via promoting autophagy, the remaining rats were randomly divided into another four groups: (1) Sham + NS; (2) CLP + NS; (3) CLP + Clemastine Fumarate; and (4) CLP + Clemastine Fumarate + 3-MA group.

Dosage form

Applications

References:
[1] Meng S, Sun X, Juan Z, et al. Clemastine Fumarate Attenuates Myocardial Ischemia Reperfusion Injury Through Inhibition of Mast Cell Degranulation. Front Pharmacol. 2021 Aug 27;12:704852.
[2] Wang X, Xie D, Dai H, et al. Clemastine protects against sepsis-induced myocardial injury in vivo and in vitro. Bioengineered. 2022 Mar;13(3):7134-7146.