摘要
目的:探讨姜黄素通过调控TLRs/NF-κB信号通路对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠的抗凋亡作用及其机制。方法:将45只C57BL/6雌性小鼠随机分为空白组、EAE组、姜黄素组,每组15只。空白组不做处理。EAE组及姜黄素组通过经典造模法建立EAE模型。自造模当日起空白组与EAE组腹腔注射生理盐水1mL·kg^(-1)·d^(-1),姜黄素组予以100mg·kg^(-1)·d^(-1)连续腹腔注射姜黄素提取物。以Benson 5评分标准每天对小鼠进行神经功能障碍评分,EAE组及姜黄素组小鼠于发病高峰期进行处死取材,空白组和其余未发病小鼠在饲养4周后处死并取出脊髓组织,取腰膨大段组织,通过HE染色、TUNEL染色观察姜黄素对EAE小鼠脊髓组织病理变化的影响以并计算其凋亡细胞数表达;免疫荧光双标染色观察凋亡相关蛋白Bcl-2、Bax与脊髓组织中凋亡细胞分布情况及共聚集现象;WesternBlot检测各组小鼠脊髓组织TLR4、NF-κBp65、MyD88蛋白表达水平。结果:与空白组相比,EAE组TUNEL染色提示脊髓组织凋亡细胞数增加(P<0.05),凋亡相关蛋白Bcl-2的表达下降,Bax的表达上升,Westernblot检测可见小鼠脊髓组织TLR4、NF-κBp65、MyD88蛋白表达水平上升(P<0.05);与EAE组相比,姜黄素组TUNEL染色脊髓组织凋亡细胞数减少(P<0.05),凋亡相关蛋白Bcl-2的表达上升,Bax蛋白表达下降,Westernblot检测可见小鼠脊髓组织TLR4、NF-κBp65、MyD88蛋白表达水平下降(P<0.05)。结论:姜黄素对EAE小鼠具有抗凋亡作用,其机制可能与其抑制TLRs/NF-κB信号通路,从而减少凋亡蛋白产生有关。
Objective:To investigate the mechanism and anti-apoptosis effect of curcumin(cur)in experimental autoim-mune encephalomyelitis(EAE)mice by regulating TLRs/NF-κB signaling pathway.Methods:In total of 45 C57BL/6 mice were randomly divided into the control group,the EAE group,the curcumin group,with 15 mice in each group.The EAE model was established by classical modeling method in the EAE group and the curcumin group.From the day of modeling,the control group and the EAE group wereintraperitoneally injected with 1mL·kg^(-1)·d^(-1) ofnormalsaline,and the curcumin group was given100mg·kg^(-1)·d^(-1) intraperitoneal injection of curcumin extract.Benson 5 was used to assess the dysneuria of the mice each day.Mice in the EAE group and curcumin group were sacrificed at the peak period of the disease.Mice in the control group and the rest of the mice were sacrificed after 4 weeks of feeding,and the spinal cord tissue was taken out to separate the lumbar enlargement segment.The effects of curcumin on the pathological changes of spinal cord tissue in EAE mice were observed by HE staining and TUNEL staining,and the expression of apoptosis positive cells was calculated.The distribution and co-aggregation of apoptosis related proteins Bcl-2 and Bax,spinal cord tissue were observed by double immunofluorescence staining.The protein levels of TLR4,NF-κBp65 and MyD88 were detected by Western blot.Results:Compared with the control group,TUNEL staining showed higher number of apoptotic cells(P<0.05),and the expression of apoptosis related protein Bcl-2 decreased and the expression of Bax increased in the EAE group(P<0.05);Beisdes,result of Western bloting showed that the protein of TLR4,NF-κBp65 and MyD88 in spinal cord tissue of mice increased in the EAE group(P<0.05).Compared with the EAE group,the number of apoptotic cells in spinal cord tissue of the curcumin group decreased(P<0.05);The expression of apoptosis related protein Bcl-2 increased,while the expression of Bax protein was decreased.Western blot result showed that the protein of TLR4,NF-κBp65 and MyD88 in spinal cord tissue of mice decreased in the curcumin group(P<0.05).Conclusion:Curcumin has anti-apoptotic effect on EAE mice,and its mechanism may be related to the inhibition of TLRs/NF-κB signaling pathway and the reduction of apoptotic protein production.
作者
刘梦兰
谢阳
曾春蓉
李作孝
LIU Meng-lan;XIE Yang;ZENG Chun-rong;LI Zuo-xiao(The Affiliated Hospital of Southwest Medical University,Luzhou 646000,China)
出处
《海南医学院学报》
CAS
2021年第20期1527-1533,共7页
Journal of Hainan Medical University
基金
泸州市人民政府基金课题(2018LZXNYD-ZK17)。
