Zhou Xin-Yu, Cui Yang, Sun Zhong-Ren, Zhang Shuo, Zhu Jia-Min, Wang Yu-Xin, Zhong Si-Tong, Yang Juan-di, Yin Hong-Na
Heilongjiang University of Chinese Medicine, Harbin 150040, China.
Second Affiliated Hospital of Heilongjiang University of Chinese Medicine, Harbin 150001.
Zhen Ci Yan Jiu. 2024 Nov 25;49(11):1129-1137. doi: 10.13702/j.1000-0607.20230902.
To observe the effect of electroacupuncture (EA) of "Jiaji" (EX-B2) on expression of high mobility group box 1 (HMGB1)/Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) related proteins and inflammation-related factors in spinal cord injury (SCI) rats, so as to explore the dynamic process of inhibiting inflammatory response in rats with SCI, and to provide a new idea and theoretical basis of molecular biology for the treatment of SCI.
Fifty-four SD rats were randomly divided into sham operation, model, and EA groups (=18 in each group), which were further divided into 3 d, 7 d and 14 d subgroups, with 6 rats at each time point. The SCI model was established according to the Allen's method. Rats in EA group received EA (1 mA, 100 Hz) intervention at EX-B2 of T9 and T11 24 hours after modeling for 30 min, once a day for 3 d, 7 d, or 14 d, respectively. The motor function of rats' hind limbs was evaluated using BBB scale, the morphological structure of rats' spinal cord tissue was observed by H.E. staining. The contents of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in spinal cord tissue were detected by ELISA. Western blot and immunofluorescence staining were used to detect the relative protein expression and fluorescence positive expression of HMGB1, TLR4 and NF-κB p65 in spinal cord tissue, respectively.
Compared with the sham operation group, the BBB scores after modeling and on day 3, 7 and 14 were all decreased (<0.05), while the contents of iNOS and COX-2 in spinal cord tissue, the protein expression and fluorescence positive expression of HMGB1, TLR4, and NF-κB p65 were increased (<0.05) in the model group. In comparison with the model group, the BBB scores on day 3, 7 and 14 were obviously increased (<0.05), while the contents of iNOS and COX-2, the protein expression and fluorescence positive expression of HMGB1, TLR4, and NF-κB p65 were significantly down-regulated in the EA group (<0.05).
EA of "Jiaji" can promote the recovery of SCI neurological function, which may be related to its function in regulating the HMGB1/TLR4/NF-κB signaling pathway, down-regulating the expression of iNOS and COX-2, and inhibiting the neuroinflammatory response after SCI.
观察电针“夹脊”（EX - B2）对脊髓损伤（SCI）大鼠高迁移率族蛋白B1（HMGB1）/Toll样受体4（TLR4）/核因子κB（NF -κB）相关蛋白及炎症相关因子表达的影响，以探讨其抑制SCI大鼠炎症反应的动态过程，为SCI的治疗提供分子生物学新思路及理论依据。
将54只SD大鼠随机分为假手术组、模型组和电针组（每组18只），每组再分为3 d、7 d和14 d亚组，每个时间点6只大鼠。采用Allen法建立SCI模型。电针组大鼠于造模后24小时在T9和T11夹脊穴接受电针干预（1 mA，100 Hz），每次30分钟，分别连续干预3 d、7 d或14 d。采用BBB评分法评估大鼠后肢运动功能，通过苏木精-伊红（H.E.）染色观察大鼠脊髓组织的形态结构。采用酶联免疫吸附测定（ELISA）法检测脊髓组织中诱导型一氧化氮合酶（iNOS）和环氧化酶-2（COX-2）的含量。分别采用蛋白质免疫印迹法（Western blot）和免疫荧光染色法检测脊髓组织中HMGB1、TLR4和NF-κB p65的相对蛋白表达和荧光阳性表达。
与假手术组相比，模型组造模后及术后3 d、7 d和14 d的BBB评分均降低（<0.05），脊髓组织中iNOS和COX-2的含量、HMGB1、TLR4和NF-κB p65的蛋白表达及荧光阳性表达均升高（<0.05）。与模型组相比，电针组术后3 d、7 d和14 d的BBB评分明显升高（<0.05），脊髓组织中iNOS和COX-2的含量、HMGB1、TLR4和NF-κB p65的蛋白表达及荧光阳性表达均显著下调（<0.05）。
电针“夹脊”可促进SCI神经功能恢复，其机制可能与调节HMGB1/TLR4/NF-κB信号通路、下调iNOS和COX-2表达、抑制SCI后的神经炎症反应有关。
