Male Sprague Dawley rats (180 ± 10 g) were supplied by Qinglongshan Laboratory Animal Company (Nanjing, Jiangsu, China). The animals were housed in a specific pathogen free (SPF) laboratory with standard environmental conditions. All the rats were fed with sterile rodent chow and water ad libitum. After 7 days acclimatization, AIA was induced in all the rats except for normal healthy controls. Prior to this procedure, heating-inactivated BCG was carefully grinded in IFA, and subsequently homogenized with the same volume of water to obtain complete Freund’s adjuvant (CFA, 20 mg/ml). On day 0, an intradermal injection of 0.1 ml CFA was carried out on the right hind paw, which was followed by a boost CFA injection at the base of tail 7 days later. Thereafter, the rats were divided into three groups with six rats each, and received different oral treatments:
Group 1: Normal healthy control (normal) administered with 0.5% sodium carboxymethyl cellulose solution (CMC-Na); Group 2: AIA model control (AIA) administered with 0.5% CMC-Na; Group 3: MAN-treated AIA rats (MAN + AIA) administered with MAN (dispersed in 0.5% CMC-Na by the aid of ethanol and tween-80).
The therapeutic dose of MAN was designated as 50 mg/kg, as it can result in effective concentrations in both blood and tissues (Xu et al., 2017 (link)). Also, MAN at this dose can effectively cure experimental arthritis (Yang et al., 2019 (link)). The treatment duration was 30 days, and body weight of all the rats was periodically recorded. All the animal experiment procedures were performed in accordance with the national institutes of health guide for the care and use of laboratory animals (NIH Publications No. 8023, revised 1978) and the approval for animal studies in this study was obtained from the Ethics Committee of Wannan Medical College.
Group 1: Normal healthy control (normal) administered with 0.5% sodium carboxymethyl cellulose solution (CMC-Na); Group 2: AIA model control (AIA) administered with 0.5% CMC-Na; Group 3: MAN-treated AIA rats (MAN + AIA) administered with MAN (dispersed in 0.5% CMC-Na by the aid of ethanol and tween-80).
The therapeutic dose of MAN was designated as 50 mg/kg, as it can result in effective concentrations in both blood and tissues (Xu et al., 2017 (link)). Also, MAN at this dose can effectively cure experimental arthritis (Yang et al., 2019 (link)). The treatment duration was 30 days, and body weight of all the rats was periodically recorded. All the animal experiment procedures were performed in accordance with the national institutes of health guide for the care and use of laboratory animals (NIH Publications No. 8023, revised 1978) and the approval for animal studies in this study was obtained from the Ethics Committee of Wannan Medical College.
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