四氢大麻酚对C57BL/6小鼠自身免疫性肝炎的保护作用
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摘要
为研究大麻素成分[四氢大麻酚(tetrahydrocannabinol, THC)]对C57BL/6小鼠自身免疫性肝炎(autoimmune hepatitis, AIH)的影响,尾静脉注射质粒pCYP2D6和pcDNA3.1,腹腔重复注射四氯化碳(carbon tetrachloride, CCl_4)和THC,ELISA检测自身免疫应答程度,微板法检测血清中谷丙转氨酶(alanine aminotransferase, ALT)活性,HE染色检测肝脏炎症反应程度,天狼猩红染色检测肝脏纤维化程度。结果显示,THC对由CCl_4/CYP2D6引起的AIH造成的肝损伤有显著改善,而且对自身免疫应答有显著抑制作用,自身抗体和抗CYP2D6抗体水平显著降低,对于单独由CYP2D6引起的抗CYP2D6抗体能够完全抑制。THC对AIH和肝纤维化也有显著的抑制作用,且THC的抑制作用与其浓度有关。由此THC对由CCl_4/CYP2D6引起的C57BL/6小鼠的AIH有显著抑制作用,并且呈剂量依赖性。由此大麻素类药物THC对AIH可能有保护作用。
To investigate the effect of cannabinoids(tetrahydrocannabinol, THC) on autoimmune hepatitis in C57BL/6 mice, the plasmids pCYP2D6 and pcDNA3.1 were injected intravenously and the carbon tetrachloride(CCl_4) and THC were repeatedly injected intraperitoneally. The autoimmune response was detected in vivo by ELISA. The serum alanine aminotransferase(ALT) enzyme activity was measured. HE staining was used to detect liver inflammation. Liver fibrosis was detected by Sirius red staining. The results showed that THC had significant inhibitory effect on liver damage and autoimmune responses caused by CCl_4/CYP2D6. Autoantibodies and anti-CYP2D6 antibodies were significantly reduced by THC. Anti-CYP2D6 antibody caused by CYP2D6 alone was completely inhibited by THC. THC also had significant inhibitory effect on autoimmune hepatitis and liver fibrosis, and the inhibitory effect depended on its concentrations. Thus, THC has a significant dose-dependent inhibitory effect on autoimmune hepatitis caused by CCl_4/CYP2D6 in C57BL/6 mice, and could have a therapeutic effect on autoimmune hepatitis.
To investigate the effect of cannabinoids(tetrahydrocannabinol, THC) on autoimmune hepatitis in C57BL/6 mice, the plasmids pCYP2D6 and pcDNA3.1 were injected intravenously and the carbon tetrachloride(CCl_4) and THC were repeatedly injected intraperitoneally. The autoimmune response was detected in vivo by ELISA. The serum alanine aminotransferase(ALT) enzyme activity was measured. HE staining was used to detect liver inflammation. Liver fibrosis was detected by Sirius red staining. The results showed that THC had significant inhibitory effect on liver damage and autoimmune responses caused by CCl_4/CYP2D6. Autoantibodies and anti-CYP2D6 antibodies were significantly reduced by THC. Anti-CYP2D6 antibody caused by CYP2D6 alone was completely inhibited by THC. THC also had significant inhibitory effect on autoimmune hepatitis and liver fibrosis, and the inhibitory effect depended on its concentrations. Thus, THC has a significant dose-dependent inhibitory effect on autoimmune hepatitis caused by CCl_4/CYP2D6 in C57BL/6 mice, and could have a therapeutic effect on autoimmune hepatitis.
引文
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[13]Hegde VL,Hegde S,Cravatt BF,et al.Attenuation of experimental autoimmune hepatitis by exogenous and endogenous cannabinoids:Involvement of regulatory T cells[J].Mol Pharmacol,2008,74(1):20-33.
[2]Manns MP,Strassburg CP.Autoimmune hepatitis:Clinical challenges[J].Gastroenterology,2001,120(6):1502-1517.
[3]Sebode M,Hartl J,Vergani D,et al.Autoimmune hepatitis:From current knowledge and clinical practice to future research agenda[J].Liver Int,2018,38(1):15-22.
[4]Li XG,Kaminski NE,Fischer LJ.Examination of the immunosuppressive effect of delta(9)-tetrahydrocannabinol in streptozotocin-induced autoimmune diabetes[J].Int Immunopharmacol,2001,1(4):699-712.
[5]Dalton GD,Howlett AC.Cannabinoid CB1receptors transactivate multiple receptor tyrosine kinases and regulate serine/threonine kinases to activate ERK in neuronal cells[J].Br J Pharmacol,2012,165(8,SI):2497-2511.
[6]Ribeiro R,Yu F,Wen J,et al.Therapeutic potential of a novel cannabinoid agent CB52in the mouse model of experimental autoimmune encephalomyelitis[J].Neuroscience,2013,254:427-442.
[7]Ismail M,Hasan H,El-Orfali Y,et al.Anti-inflammatory,antioxidative,and hepatoprotective effects of transΔ9-tetrahydrocannabinol/sesame oil on adjuvant-induced arthritis in rats[J].Evid Based Complement Alternat Med,2018:9365464.
[8]Manns MP,Lohse AW,Vergani D.Autoimmune hepatitisUpdate 2015[J].J Hepatol,2015,62(1):S100-S111.
[9]Christen U.Animal models of autoimmune hepatitis[J].Biochim Biophys Acta Mol Basis Dis,2019,1865(5):970-981.
[10]Chi G,Feng XX,Ru YX,et al.TLR2/4ligand-amplified liver inflammation promotes initiation of autoimmune hepatitis due to sustained IL-6/IL-12/IL-4/IL-25expression[J].Mol Immunol,2018,99:171-181.
[11]Harada K,Hiep NC,Ohira H.Challenges and difficulties in pathological diagnosis of autoimmune hepatitis[J].Hepatol Res,2017,47(10):963-971.
[12]De Lago E,Moreno-Martet M,Cabranes A,et al.Cannabinoids ameliorate disease progression in a model of multiple sclerosis in mice,acting preferentially through CB1receptormediated anti-inflammatory effects[J].Neuropharmacology,2012,62(7):2299-2308.
[13]Hegde VL,Hegde S,Cravatt BF,et al.Attenuation of experimental autoimmune hepatitis by exogenous and endogenous cannabinoids:Involvement of regulatory T cells[J].Mol Pharmacol,2008,74(1):20-33.