摘要
目的探究3-乙酰基-11-酮基-β-乳香酸(AKBA)对放射性脊髓损伤(RI-SCI)的治疗作用及其机制。方法HT22细胞购自上海富衡生物科技有限公司。未处理细胞为NC组,照射后为RT组,照射后加AKBA处理为RA组,将转染的阴性对照、核因子E2相关因子2(NRF2)的小干扰RNA分别转染至细胞中,照射后加入AKBA,分为RA+si-NC组和RA+si-NRF2组,30只SD大鼠采用随机数字表法分为3组:未处理为Control组(10例)、照射后为IR组(10例),照射后AKBA处理为Treat组(10例)。细胞计数试剂盒(CCK-8)检测细胞活性;记录大鼠照射后瘫痪潜伏期,苏木精-伊红(HE)染色观察脊髓组织病理变化,蛋白质印迹法(Western blot)检测NRF2、血红素氧合酶1(HO-1)、白细胞介素(IL)-1β、IL-6蛋白的表达,荧光定量聚合酶链反应(PCR)检测脊髓组织中NRF2、HO-1的mRNA水平,计算分子对接的结合能评估AKBA和NRF2的结合状态。组间比较采用t检验。结果RA组细胞活性高于RT组(0.74±0.03比0.49±0.08,t=4.990、5.789,P<0.05),RA+si-NRF2组与RT+si-NRF2组之间差异无统计学意义(0.39±0.10比0.31±0.03,t=1.368,P>0.05),RT组IL-1β、IL-6蛋白高于RA组(1.16±0.11比0.49±0.03、1.35±0.18比0.92±0.04,t=10.565、4.061,P<0.05),RA组NRF2、HO-1蛋白高于RT组(1.10±0.05比0.84±0.09、0.93±0.03比0.82±0.03,t=4.498、4.450,P<0.05);RA+si-NRF2组IL-1β蛋白显著高于RA+si-NC组(0.86±0.03比0.51±0.11,t=5.345,P<0.05)。Treat组瘫痪潜伏期长于IR组[(133.20±3.90)d比(121.60±3.58)d,χ^(2)=21.54,P<0.05],IR组IL-1β、IL-6高于Treat组(1.15±0.17比0.57±0.10、1.31±0.16比0.47±0.12,t=4.983、7.173,P<0.05),Treat组NRF2、HO-1蛋白、mRNA高于IR组(1.11±0.14比0.60±0.17、1.09±0.12比0.43±0.10、10.41±0.65比6.78±0.34、50.61±5.45比12.84±2.22,t=4.060、7.311、11.067、14.361,P<0.05)。分子对接结合能为-11.36 kcal/mol。结论AKBA通过激活NRF2/HO-1信号通路抑制脊髓炎性反应,从而保护脊髓神经元,改善RI-SCI。
Objective To explore the therapeutic effect of 3-acetyl-11-keto-β-boswellia acid on radiation-induced spinal cord injury and its mechanism.Methods HT22 cell was purchased from Shanghai Fuheng Biotechnology Co.Cells after irradiation were RT group,and RA group after irradiation with AKBA treatment.Negative control and nuclear factor erythroid2-related factor 2(NRF2)small interfering RNA were transfected into irradiated cells,respectively.AKBA was added and divided into RA+si-NC group and RA+si-NRF2 group.30 SD rats were divided into 3 groups using the random number table method:Control group(n=10),irradiated as IR group(n=10),and AKBA treated after irradiation as Treat group(n=10).Cell counting kit(CCK-8)was use to detect cell proliferation and hematoxylin and eosin(HE)staining of spinal cord tissue were performed;Record the latency of paralysis after irradiation,and detect NRF2,HO-1(Heme Oxygenase-1),Interleukin(IL)-6,and IL-1βprotein with Western blotting.The mRNA levels of NRF2 and HO-1 in spinal cord tissue were detected by fluorescence quantitative polymerase chain reaction.The binding state of AKBA and NRF2 was evaluated by calculating the binding energy of molecular docking.The t-test was used for comparison between groups.Results Cell activity of the RA group was higher than that RT group(0.74±0.03 vs.0.49±0.08,t=4.990,5.789,P<0.05)and there was no significant difference between the RA+si-NRF2 group and the RT+si-NRF2 group(0.39±0.10 vs.0.31±0.03,t=1.368,P>0.05),IL-1βand IL-6 proteins in the RT group were higher than RA group(1.16±0.11 to 0.49±0.03,1.35±0.18 to 0.92±0.04,t=10.565,4.061,P<0.05).NRF2 and HO-1 proteins in the RA group were higher than RT group(1.10±0.05 vs.0.84±0.09,0.93±0.03 vs.0.82±0.03,t=4.498,4.450,P<0.05).IL-1βlevels of RA+si-NRF2 group were higher than RA+si-NC group(0.86±0.03 vs.0.51±0.11,t=5.345,P<0.05).The latency of paralysis in Treat group was longer than IR group[(133.20±3.90)d vs.(121.60±3.58)d,χ^(2)=21.54,P<0.05],IL-1βand IL-6 protein in IR group were highe
作者
吴寒
钱泽宇
周志强
崔红霞
孙彦泽
畅磊
张鹏
Wu Han;Qian Zeyu;Zhou Zhiqiang;Cui Hongxia;Sun Yanze;Chang Lei;Zhang Peng(Department of Orthopedics,the Second Affiliated Hospital of Soochow University,Suzhou 215004,China;Department of Pathology,the Second Affiliated Hospital of Soochow University,Suzhou 215004,China;Department of Radiotherapy,the Second Affiliated Hospital of Suzhou University,Soochow 215004,China;State Key Laboratory of Radiation Medicine and Protection of Soochow University,Suzhou 215123,China)
出处
《中华实验外科杂志》
CAS
北大核心
2023年第12期2559-2562,共4页
Chinese Journal of Experimental Surgery
基金
省部共建放射医学与辐射防护国家重点实验室开放课题(GZK1202128)
苏州市中西医结合科研基金(SKJYD2021220)。
