摘要
目的:观察黄芪汤对^(12)C^(6+)辐射脑损伤大鼠下丘脑组织中miRNA的基因表达及TLR4/MyD88/NF-κB信号通路相关蛋白表达的影响,初步探讨黄芪汤对重离子辐射损伤的防护机制。方法:SPF级Wistar大鼠50只随机分为正常对照组,辐射模型组,黄芪汤高、中、低剂量组(18、9、4.5 g·kg^(-1)·d^(-1)),灌胃2周。干预7 d后除正常对照组外,其余各组脑组织给予4 Gy^(12)C^(6+)离子束单次照射,辐射后7 d处死各组大鼠。HE染色观察垂体组织的病理形态学改变;ELISA检测下丘脑组织IL-1β、IL-6的分泌情况;RT-PCR法检测下丘脑中miRNA与TLR4的基因表达;Western blot法检测下丘脑中TLR4、MyD88、NF-κB p65、p-NF-κB p65蛋白的表达。结果:与辐射模型组比较,黄芪汤高剂量组IL-1β水平显著降低(P<0.01),中、高剂量组IL-6水平及miR-93、TLR4的基因表达显著降低(P<0.01),miR-140-5p的基因表达显著增加(P<0.01),黄芪汤各剂量组中TLR4、MyD88、NF-κB p65以及p-NF-κB p65蛋白表达显著降低(P<0.01),黄芪汤高剂量组核固缩、深染现象明显减轻,偶见凋亡小体。Pearson分析表明,下丘脑中miR-93与TLR4蛋白、mRNA表达水平具有显著正线性相关性(P<0.01),而miR-140-5p与TLR4蛋白和mRNA表达水平呈现负线性相关性(P<0.01)。结论:黄芪汤可能是通过抑制miR-93的过度表达和促进miR-140-5p的表达抑制TLR4/MyD88/NF-κB信号通路的激活,从而抑制其下游的炎症级联反应,减轻重离子辐射后造成的下丘脑损伤,发挥抗辐射的作用。
Objective:To observe the effects of Huangqi Decoction on the expression of miRNA genes in the hypothalamus and the expression of related proteins in the TLR4/MyD88/NF-κB signaling pathway in^(12)C^(6+)radiation brain injury model rats,and to preliminarily explore the protective mechanism of Huangqi Decoction on heavy ion radiation damage.Methods:Fifty SPF Wistar rats were randomly divided into normal control group,radiation model group,Huangqi Decoction high,medium and low dose groups (18,9,4.5 g·kg^(-1)·d^(-1)) were administered orally for 2 weeks.After 7 days of intervention,except for the normal control group,the brain tissues of the other groups were irradiated with 4 Gy^(12)C^(6+)ion beam for a single time,and the rats in each group were sacrificed 7 days after the irradiation.HE staining to observe the pathological changes of pituitary tissue;ELISA to detect the secretion of IL-1β and IL-6 secretion in the hypothalamic tissue;RT-PCR method to detect the gene expression of miRNA and TLR4 in the hypothalamus;Western blot method to detect TLR4,MyD88,NF-κB p65,p-NF-κB p65 protein expression in the hypothalamus.Results:Compared with the radiation model group,the level of IL-1β in the high-dose group of Huangqi Decoction decreased (P<0.01);the level of IL-6 and the gene expression of miR-93 and TLR4 decreased significantly(P<0.01),while the gene expression of miR-140-5p in the medium and high dose groups was significantly increased (P<0.01).The protein expression of TLR4,MyD88,NF-κB p65 and p-NF-κB p65 in Huangqi Decoction group was significantly reduced(P<0.01).The high-dose Huangqi decoction group significantly reduced nuclear pyknosis and deep staining,and occasionally apoptotic bodies.Pearson analysis showed that the expression levels of miR-93 and TLR4 protein and mRNA in the hypothalamus had a significant positive linear correlation (P<0.01),while the protein and mRNA expression levels of miR-140-5p and TLR4showed negative linearity correlation (P<0.01).Conclusion:Huangqi Decoction may inhibit the
作者
蒋国凤
颜春鲁
安方玉
刘永琦
石瑶
吕栋辉
袁灵青
李孟翰
王嘉玉
JIANG Guo-feng;YAN Chun-lu;AN Fang-yu;LIU Yong-qi;SHI Yao;LYU Dong-hui;YUAN Ling-qing;LI Meng-han;WANG Jia-yu(Gansu University of Chinese Medicine,Lanzhou 730101,China;Gansu Provincial Research Center of Traditional Chinese Medicine,Lanzhou 730000,China;Gansu Provincial Key Laboratory of Chronic Diseases for Prevention and Treatment of Traditional Chinese Medicine,Lanzhou 730000,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2023年第2期805-810,共6页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81760797)
甘肃省“双一流”科研重点项目(No.GSSYLXM-05)
甘肃省优秀研究生“创新之星”项目(No.2021CXZX-746)
甘肃中医药大学科学研究与创新基金(No.2021KCYB-4)。
