Landucci Elisa, Mazzantini Costanza, Lana Daniele, Giovannini Maria Grazia, Pellegrini-Giampietro Domenico E
Department of Health Sciences, Section of Clinical Pharmacology and Oncology, University of Florence, Viale Pieraccini 6, 50139 Florence, Italy.
Toxics. 2022 Jan 21;10(2):48. doi: 10.3390/toxics10020048.
Cannabis derivatives are largely used in the general population for recreational and medical purposes, with the highest prevalence among adolescents, but chronic use and abuse has raised medical concerns. We investigated the prolonged effects of Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD) in organotypic hippocampal slices from P7 rats cultured for 2 weeks. Cell death in the CA1 subregion of slices was quantified by propidium iodide (PI) fluorescence, pre-synaptic and post-synaptic marker proteins were analysed by Western blotting and neurodegeneration and astrocytic alterations by NeuN and GFAP by immunofluorescence and confocal laser microscopy. The statistical significance of differences was analysed using ANOVA with a post hoc Dunnett w-test (PI fluorescence intensities and Western blots) or Newman-Keuls (immunohistochemistry data) for multiple comparisons. A probability value () of < 0.05 was considered significant. Prolonged (72 h) THC or CBD incubation did not induce cell death but caused modifications in the expression of synaptic proteins and morphological alterations in neurons and astrocytes. In particular, the expression of PSD95 was reduced following incubation for 72 h with THC and was increased following incubation with CBD. THC for 72 h caused disorganisation of CA1 stratum pyramidalis (SP) and complex morphological modifications in a significant number of pyramidal neurons and in astrocytes. Our results suggest that THC or CBD prolonged exposure induce different effects in the hippocampus. In particular, 72 h of THC exposure induced neuronal and glia alterations that must draw our attention to the effects that relatively prolonged use might cause, especially in adolescents.
大麻衍生物在普通人群中大量用于娱乐和医疗目的，在青少年中使用率最高，但长期使用和滥用引发了医学关注。我们研究了Δ9-四氢大麻酚（THC）和大麻二酚（CBD）对培养2周的P7大鼠海马脑片的长期影响。通过碘化丙啶（PI）荧光定量脑片CA1亚区的细胞死亡，通过蛋白质免疫印迹分析突触前和突触后标记蛋白，通过免疫荧光和共聚焦激光显微镜用NeuN和GFAP分析神经变性和星形细胞改变。使用方差分析（ANOVA）及事后Dunnett w检验（PI荧光强度和蛋白质免疫印迹）或Newman-Keuls检验（免疫组织化学数据）分析差异的统计学意义，用于多重比较。概率值（）<0.05被认为具有统计学意义。THC或CBD长时间（72小时）孵育未诱导细胞死亡，但导致突触蛋白表达改变以及神经元和星形细胞的形态改变。特别是，与THC孵育72小时后PSD95的表达降低，与CBD孵育后PSD95的表达增加。THC处理72小时导致CA1锥体层（SP）紊乱，并在大量锥体神经元和星形细胞中引起复杂的形态学改变。我们的结果表明，THC或CBD长时间暴露在海马体中会产生不同的影响。特别是，72小时的THC暴露会引起神经元和神经胶质细胞的改变，这必须引起我们对相对长时间使用可能产生的影响的关注，尤其是在青少年中。
