|
| LL-37抗菌肽对慢性炎性疼痛大鼠炎症和氧化应激的影响 |
| Effects of LL-37 antimicrobial peptide on inflammation and oxidative stress in rats with chronic inflammatory pain |
| |
| DOI：10.12089/jca.2026.02.012 |
| 中文关键词: LL-37抗菌肽 炎性疼痛 小胶质细胞 Toll样受体4 核因子κB |
| 英文关键词: LL-37 antimicrobial peptide Chronic inflammatory pain Microglia Toll-like receptor 4 Nuclear factor kappa-light-chain-enhancer of activated B cells |
| 基金项目:湖南省人民医院医联体专项科研基金项目（2023YLT010） |
|
| 摘要点击次数: 823 |
| 全文下载次数: 254 |
| 中文摘要: |
目的：探讨LL-37抗菌肽对慢性炎性疼痛（CIP）大鼠炎症和氧化应激的影响，以及背根神经节（DRG）组织中Toll样受体4（TLR4）/核因子κB（NF-κB）信号通路的机制作用。
方法：选择SPF级SD大鼠36只，8周龄，体重190～210 g。采用随机数字表法将大鼠分为三组：对照组（Con组）、模型组（CIP组）和模型+LL-37抗菌肽组（LL37组），每组12只。Con组为正常大鼠，鞘内注射生理盐水10 μl，每日1次，连续注射7 d；CIP组于CIP大鼠模型建立成功次日，鞘内注射生理盐水10 μl，每日1次，连续注射7 d；LL37组于CIP大鼠模型建立成功次日，鞘内注射LL-37抗菌肽2 mg/kg，每日1次，连续注射7 d。于药物干预前1 d、药物干预后3、5、7 d进行疼痛行为学实验，测定机械性缩足反射阈值和热缩足反射潜伏期。后处死大鼠，分离DRG组织，采用ELISA法检测白细胞介素（IL）-1β、IL-6、肿瘤坏死因子-α（TNF-α）、活性氧（ROS）、丙二醛（MDA）和超氧化物歧化酶（SOD）浓度。采用Western blot法检测TLR4、NF-κB p65、诱导型一氧化氮合酶（iNOS）、CD86、CD206、IL-10蛋白含量。采用免疫荧光染色记录Iba-1阳性细胞数量。
结果：与Con组比较，CIP组术后3、5、7 d机械性缩足反射阈值明显降低、热缩足反射潜伏期明显缩短，IL-1β、IL-6、TNF-α、ROS和MDA浓度明显升高，SOD浓度明显降低，TLR4、NF-κB p65、iNOS和CD86蛋白含量明显升高，CD206和IL-10蛋白含量明显降低，Iba-1阳性细胞数量明显增加（P<0.05）。与CIP组比较，LL37组术后3、5、7 d机械性缩足反射阈值明显升高、热缩足反射潜伏期明显延长，IL-1β、IL-6、TNF-α、ROS和MDA浓度明显降低，SOD浓度明显升高，TLR4、NF-κB p65、iNOS和CD86蛋白含量明显降低，CD206和IL-10蛋白含量明显升高，Iba-1阳性细胞数量明显减少（P<0.05）。
结论：LL-37抗菌肽通过诱导DRG组织中小胶质细胞由M1向M2型极化，抑制炎症及氧化应激，从而减轻CIP，其作用机制可能与抑制TLR4/NF-κB信号通路相关。 |
| 英文摘要: |
Objective: To investigate the effects of LL-37 antimicrobial peptide on inflammation and oxidative stress in rats with chronic inflammatory pain (CIP), and the mechanism of Toll-like receptor 4 (TLR4) / nuclear factor-κB (NF-κB) signaling pathway in dorsal root ganglion (DRG).
Methods: Thirty-six SPF grade SD rats, 8 weeks old, weighing 190-210 g, were randomly divided into three groups: control group (group Con), model group (group CIP), and model + LL-37 antimicrobial peptide group (group LL37), 12 rats in each group. Group Con was normal rats, intrathecal injection of normal saline 10 μl, once a day for 7 days. Group CIP got intrathecal injection of normal saline 10 μl 1 day after CIP model was successfully established, once a day for 7 days. Group LL37 got intrathecal injection of LL-37 antimicrobial peptide 2 mg/kg 1 day after CIP model was successfully established, once a day for 7 days. Pain behavioral experiments were performed 1 day before drug intervention, 3, 5, and 7 days after drug intervention to determine the mechanical paw withdrawal threshold and thermal paw withdrawal latency. The rats were sacrificed and DRG tissues were isolated. The concentrations of interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF-α), reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD) were detected by ELISA. The protein levels of TLR4, NF-κB p65, inducible nitric oxide synthase (iNOS), CD86, CD206, and IL-10 were detected by Western blot. The number of Iba-1 positive cells was recorded by immunofluorescence staining.
Results: Compared with group Con, the mechanical withdrawal threshold was significantly decreased, the thermal withdrawal latency was significantly shortened, the concentrations of IL-1β, IL-6, TNF-α, ROS, and MDA were significantly increased, the concentration of SOD was significantly decreased, the contents of TLR4, NF-κB p65, iNOS, and CD86 proteins were significantly increased, the contents of CD206 and IL-10 proteins were significantly decreased, and the number of Iba-1 positive cells was significantly increased 3, 5, and 7 days after operation in group CIP (P < 0.05). Compared with group CIP, the mechanical withdrawal threshold was significantly increased, the thermal withdrawal latency was significantly prolonged, the concentrations of IL-1β, IL-6, TNF-α, ROS, and MDA were significantly decreased, the concentration of SOD was significantly increased, the protein levels of TLR4, NF-κB p65, iNOS, and CD86 were significantly decreased, the protein levels of CD206 and IL-10 were significantly increased, and the number of Iba-1 positive cells was significantly decreased 3, 5, and 7 days after operation in group LL37 (P < 0.05).
Conclusion: LL-37 antimicrobial peptide can inhibit inflammation and oxidative stress by inducing microglia polarization from M1 to M2 in DRG tissue, thereby reducing CIP. The mechanism may be related to the inhibition of TLR4/NF-κB signaling pathway. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
