Sickle cell anemia can cause the pain linked to an accumulation of misshapen red blood cells in the microscopic capillaries. Also, pain can occur from poor oxygenation of the tissues.
Iron deficiency anemia symptoms include: (9 )
CBD Oil and Sickle Cell Anemia
CBD has been shown to possess therapeutic benefits that might help with symptoms or conditions linked to some types of anemia.
A study used an animal model to look at how cannabinoids can be effective at treating the pain from sickle cell anemia (5 ) .
The American Society of Hematology recommends taking 150-200 mg of iron each day, or 2 to 5 mg of iron for every kilogram of body weight per day (8 ) .
The results were analyzed using SPSS 20.0 and expressed as means ± S.E.M. Levene’s test of equality of variances was used in order to test the assumption of homogeneity of variance. Variances were similar among the experimental groups for all tested variables. Statistical comparisons were performed using two-way analysis of variance (2-way-ANOVA), with neonatal treatment (vehicle or iron) and adult treatment (vehicle or CBD) as fixed factors. One-way ANOVA, followed by Tukey’s post hoc test, was used to test differences between the experimental groups. In all comparisons, p values below 0.05 were considered as indicative of statistical significance.
The present findings show that iron overload induced increases in APAF1, Caspase 3, Caspase 9, Cytochrome c, and cleaved PARP levels. Although we have not performed a direct measurement of apoptosis, increased cleaved PARP levels have been considered a marker of apoptosis because this protein is the substrate of activated caspases 43 . In agreement, upregulation of Caspase 3 gene expression in a model of cognitive impairment induced by sevoflurane was associated with increased cleaved PARP levels 44 . While Caspase 3 is an effector caspase, being part of the final common pathway of apoptosis, Cytochrome c, APAF 1, and Caspase 9 integrate the intrinsic apoptotic pathway. Interestingly, no alterations in cleaved-Caspase 8 levels were found, confirming that there was no activation of extrinsic apoptosis pathways. On the other hand, we found increases in Caspase 9, Cytochrome c and APAF1 levels, suggesting that the intrinsic pathway is most significantly affected by iron overload. Since mitochondria are the main source of ROS, they are expected to become an important target of oxidative damage, which could explain functional alterations in these organelles in pathological conditions 45 . Moreover, mitochondria play a key role in regulating the intrinsic apoptotic pathway, and there is evidence indicating that iron affects mitochondrial homeostasis 13,30 , thus supporting the concept that iron effects are most pronounced in the intrinsic pathway. Nonetheless, more studies on the effects of iron on the extrinsic pathway are warranted.
Western blotting analysis
Nowadays, many studies are being performed with CBD aiming to analyze its therapeutic properties and mechanisms of action. In this study, we showed the neuroprotective effects of the adult treatment with CBD on apoptotic markers in rats treated neonatally with iron. Considering that iron dyshomeostasis takes place throughout life, and is possibly related to an increased iron intake during early stages of life, CBD might represent a therapeutic option that ameliorates pathological processes previously initiated. We observed that adult treatment with CBD was able to rescue APAF1, Caspase 9, Caspase 3, and cleaved PARP levels. Only Cytochrome c levels were not rescued to control levels. Notwithstanding, taken together the present findings suggest that CBD was able to protect from apoptosis by reducing Caspase 3 and cleaved PARP levels, proteins that participate in the effector phase of apoptosis, which culminates in cell death. A previous study indicated that CBD attenuates the imbalance between Bcl-2/Bax, upregulating the anti-apoptotic protein Bcl-2, which in turn maintains the integrity of the outer mitochondrial membrane, in an animal model of multiple sclerosis 46 . In agreement, we have previously demonstrated that CBD protects against mitochondrial injury 13,30 , converging to prevent the activation of the intrinsic apoptotic pathway. Using a model of HD, Valdeolivas and coworkers 47 have suggested the involvement of CB1 and CB2 cannabinoid receptors as well as receptor-independent actions in the neuroprotective effects of a CBD-enriched botanical extract. Further studies are warranted in order to clarify whether CBD’s antiapoptotic effects are related or not to cannabinoid receptor agonism.
Iron accumulation in the brain has been recognized as a common feature of both normal aging and neurodegenerative diseases. Cognitive dysfunction has been associated to iron excess in brain regions in humans. We have previously described that iron overload leads to severe memory deficits, including spatial, recognition, and emotional memory impairments in adult rats. In the present study we investigated the effects of neonatal iron overload on proteins involved in apoptotic pathways, such as Caspase 8, Caspase 9, Caspase 3, Cytochrome c, APAF1, and PARP in the hippocampus of adult rats, in an attempt to establish a causative role of iron excess on cell death in the nervous system, leading to memory dysfunction. Cannabidiol (CBD), the main non-psychotropic component of Cannabis sativa, was examined as a potential drug to reverse iron-induced effects on the parameters analyzed. Male rats received vehicle or iron carbonyl (30 mg/kg) from the 12th to the 14th postnatal days and were treated with vehicle or CBD (10 mg/kg) for 14 days in adulthood. Iron increased Caspase 9, Cytochrome c, APAF1, Caspase 3 and cleaved PARP, without affecting cleaved Caspase 8 levels. CBD reversed iron-induced effects, recovering apoptotic proteins Caspase 9, APAF1, Caspase 3 and cleaved PARP to the levels found in controls. These results suggest that iron can trigger cell death pathways by inducing intrinsic apoptotic proteins. The reversal of iron-induced effects by CBD indicates that it has neuroprotective potential through its anti-apoptotic action.
We also analyzed the effects of neonatal iron loading and adult treatment with CBD on Caspases 8 and 9. Two-way ANOVA comparisons of cleaved-Caspase 8 measured by western blot revealed no significant main effects of neonatal treatment (F(1,14) = 0.13, p = 0.728, Fig. Fig.2a) 2a ) or adult treatment (F(1,14) = 0.015, p = 0.905, Fig. Fig.2a). 2a ). However, statistical comparisons of Caspase 9 levels using two-way ANOVA showed significant main effects of neonatal treatment (F(1,12) = 27.90, p < 0.0001, Fig. Fig.2b) 2b ) and adult treatment (F(1,12) = 6.79, p < 0.05, Fig. Fig.2b), 2b ), and a significant interaction (F(1,12) = 22.47, p < 0.0001, Fig. Fig.2b). 2b ). One-way ANOVA revealed significant differences in Caspase 9 protein levels among the groups (F(3, 12) = 19.05, p < 0.0001, Fig. Fig.2b). 2b ). Post hoc comparisons between groups indicated that neonatal iron treatment induced a significant increase in Caspase 9 protein levels in comparison to the control group, which received sorbitol in the neonatal period and vehicle in adulthood (p < 0.0001). The reversion effects of CBD were also observed, considering that the group that received iron in the neonatal period and CBD in adulthood (iron-CBD) showed statistically significant differences in Caspase 9 levels in comparison to the group that received iron in the neonatal period and vehicle in adult age (p = 0.001), and this group was not significantly different from the control group (p = 0.281).