## 4 Biological Activity of Cannabis Metabolites
### Biological Activity of Cannabis Alkaloids and Other Nitrogen-Containing Compounds
#### 4.2.2 In Vivo Activity of Cannabis Nitrogen-Containing Compounds  
Hordenine was tested against an ulcerative colitis mouse model by [PERSON] et al. in 2023. Mice were given a 4% dextran sodium sulphate (DSS) solution during the first 7 days to induce ulcerative colitis and Nordenine-treated mice received 12.5 mg/kg, 25 mg/kg, or 50 mg/kg from day 1 to 12 [94]. Mice were harvested on day 13. They found that mice treated with 50 mg/kg of Nordenine had a diminution in production of IL-6, IL-18, and TNF-\(\alpha\). Furthermore, the anti-inflammatory activity of Nordenine was observed through the inhibition of expression of SPHK1, SIPR1, Rac1, and p-STAT3. As well, the enhanced expression of tight junction proteins ZO-1 and occludin suggest a role in colon epithelial cell healing [94]. Hordenine was reported as a \(\beta_{2}\)-AR agonist in mice skeletal muscle by [PERSON] and [PERSON] in 2018. Hordenine (20 mg/kg) diluted in 10% DMSO in water was injected directly in mice quadriceps three times, and RNA of quadricp muscle cells was extracted six hours after injections [66]. RT-PCR was performed on the muscle extracts, and nuclear receptor-4A1 (Nr4A1) and PPAR coactivator 1 alpha 4 (PGC-1\(\alpha\)4) mRNA levels were increased when compared to the control, suggesting a potential role of hardening in muscle hypertrophic gene expression and positive skeletal muscle function. Hordenine was investigated for its anti-prolactinoma effects in rats, compared to the first-line therapy compound bromocriptine by [PERSON] et al. (2020). The rats were injected in their abdominal cavity with 17\(\alpha\)-estradiol (2 mg) to induce prolactinoma, once a day, every five days, for a total of 50 days [95]. Aqueous shoreline was then daily administered intragastrically for 30 days. Hordenine-treated rats saw a reduction in pituitary gland volume that was comparable to bromocriptine when administered with 38.2 mg/kg of hardening. Rats treated with 76.4 mg/kg and 152.8 mg/kg of hardening saw an even greater reduction in the volume of their pituitary gland than with bromocriptine. All tested concentrations of hordenine reduced prolactin serum levels, but less than bromocriptine. They concluded that hordenine seems to lessen the effects of prolactinoma through MAPK regulation, by lowering p-p38, p-ERK1/2, and p-JNK protein expression, and diminishing TNF-\(\alpha\), IL-6, and IL-18 production [95]. Similarly, hordenine's anti-inflammatory capacity was investigated in a lipopolysaccharide-induced acute lung injury (ALI) model in mice. The mouse model for ALI was established by the intranasal instillation of 1.25 mg/kg of LPS in a phosphate-buffered saline solution. Mice were pretreated with hordenine (10 mg/kg or 15 mg/kg) through intraperitoneal injections for 1 h [96]. [PERSON] et al. observed a diminution in pro-inflammatory cytokine expression, such as IL-6, IL-18, and TNF-\(\alpha\). Following the same trend, [PERSON] et al. (2022) published an article regarding hordenine's anti-inflammatory mechanism in a Parkinson's disease rat model. Neuroinflammation was induced by an injection of 6-hydroxyopamine (3 \(\mu\)L of 4 \(\mu\)g/\(\mu\)L) in the rats' right substantia nigra compact [97]. Rats were pretreated with hordenine (50 mg/kg) dissolved in distilled water, once a day for three days, by gavage, and for 5 weeks following the neuroinflammation-inducing injection. They found, through immunohistochemistry and Western blotting, that hordenine inhibits the loss of dopaminergic neurons. Furthermore, they brought evidence that hordenine's anti-neuroinflammatory activity is being mediated, at least in part, by the suppression of microglia activation, which is a considerable contributor to [PERSON]'s disease development, through TNF-\(\alpha\), IL-6, COX-2, and iNOS production, and its consequent neuroinflammation [97]. Hordenine was studied in the context of [PERSON]'s disease in rats by [PERSON] et al. (2024).