基于UPR/GRP78/IL-6信号通路探讨温通活血乳膏治疗糖尿病周围神经病变的作用机制  

Mechanism of Wentong Huoxue cream for diabetic peripheral neuropathy via UPR/GRP78/IL-6 signal pathway

作　　者：严琦 刘珊珊 古丽逊·买买提朱玛 徐利娟 马静[2] 刘新[3] 李惠[2] 王晶[4] 王先敏[2] 马丽[2] YAN Qi;LIU Shan-shan;Gulixun·Maimaitizhuma;XU Li-juan;MA Jing;LIU Xin;LI Hui;WANG Jing;WANG Xian-min;MA Li(The Fourth Clinical Medical College of XinjiangMedical University,Urumqi 830000;Hospital of Traditional Chinese Medicine Affiliated toXinjiang Medical University,Urumqi 830000;Xinjiang Medical University,Urumqi 830054;Key Laboratory of Respiratory Diseases Research,Affiliated Hospital of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi 830000)

机构地区：[1]新疆医科大学第四临床医学院,乌鲁木齐830000 [2]新疆医科大学附属中医医院,乌鲁木齐830000 [3]新疆医科大学,乌鲁木齐830054 [4]新疆医科大学附属中医医院呼吸病研究重点实验室,乌鲁木齐830000

出　　处：《中南药学》2025年第2期319-327,共9页Central South Pharmacy

基　　金：国家自然科学基金项目(No.82060842);新疆维吾尔自治区重点研发计划项目(No.2023B03002)。

摘　　要：目的探讨中药复方温通活血乳膏调控糖尿病周围神经病变(DPN)在施万细胞中内质网应激的分子机制。方法选取雄性SD大鼠建立DPN模型。分为空白组、模型组、温通活血乳膏组,连续给药28 d后评价治疗效果;并基于全转录测序技术,鉴定各组间差异表达基因。然后在高糖环境下培养大鼠施万细胞(RSC96),采用CCK-8法检测各组细胞的增殖情况,Western blot法检测UPR通路下的3种重要跨膜蛋白[蛋白激酶R样内质网激酶(PERK)、激活转录因子6(ATF6)、肌醇需求酶1(IRE1)]以及葡萄糖调节蛋白(GRP78)、凋亡蛋白C/EBP同源蛋白(CHOP)、白细胞介素(IL)-6的表达。结果与模型组相比,温通活血乳膏组可明显改善大鼠的热痛觉。透射电镜结果显示温通活血乳膏处理改善了坐骨神经的神经纤维、内质网及髓鞘的形态。此外通过全转录组测序技术发现了379个共有的差异表达基因(DEGs),共有DEGs的富集分析和蛋白-蛋白相互作用网络(PPI)显示IL-6作为核心枢纽基因在细胞因子-细胞因子受体通路上富集。CCK-8法检测结果显示25 mg·mL^(-1)温通活血乳膏能明显抑制细胞增殖(P<0.01)。Western blot实验结果表明温通活血乳膏显著降低了内质网应激通路蛋白PERK、ATF6、IRE1、GRP78和凋亡蛋白CHOP及炎症因子IL-6的表达。结论温通活血乳膏可能通过抑制UPR/GRP78/IL-6信号通路的激活,减轻施万细胞中内质网应激、细胞凋亡和炎症反应,保护DPN大鼠的坐骨神经的形态和功能,以预防及改善DPN。Objective To determine the molecular mechanism through which Wentong Huoxue cream modulates endoplasmic reticulum(ER)stress in Schwann cells associated with diabetic peripheral neuropathy(DPN).Methods Male Sprague-Dawley rats were selected to establish a DPN model.The rats were divided into three groups:a control group,a model group,and a Wentong Huoxue cream group.After 28 days of continuous treatment,the effects were evaluated.Differentially expressed genes(DEGs)among the groups were identified with whole transcriptome sequencing technology.In addition,rat Schwann cells(RSC96)were cultured in a high-glucose environment,and CCK-8 method was used to assess cell proliferation at various drug concentrations.The expression of three key transmembrane proteins in the unfolded protein response(UPR)pathway[protein kinase R-like ER kinase(PERK),activating transcription factor 6(ATF6),and inositol-requiring enzyme 1(IRE1)],along with glucose-regulated protein 78(GRP78/Bip),C/EBP homologous protein(CHOP),and interleukin-6(IL-6)was measured by Western blot.Results Compared with the model group,the Wentong Huoxue cream group obviously improved the thermal pain sensitivity.Transmission electron microscopy revealed that Wentong Huoxue cream enhanced the morphology of nerve fibers,ER,and myelin in the sciatic nerve.Whole transcriptome sequencing identified 379 shared DEGs.Enrichment analysis and protein-protein interaction network indicated that IL-6 served as a central hub gene and enriched in the cytokine-cytokine receptor pathway.CCK-8 showed that Wentong Huoxue cream at 25 mg·mL^(-1)inhibited the cell proliferation(P<0.01).Western blot showed that Wentong Huoxue cream reduced the expressions of ER stress pathway proteins PERK,ATF6,IRE1,GRP78,apoptotic protein CHOP and inflammatory factor IL-6.Conclusion Wentong Huoxue cream may alleviate ER stress,apoptosis,and inflammation in Schwann cells by inhibiting the activation of the UPR/GRP78/IL-6 signaling pathway,which helps protect the morphology and function of the sciatic ne

关 键 词：糖尿病周围神经病变 内质网应激 UPR信号通路 全转录组测序 差异表达基因 

分 类 号：R285[医药卫生—中药学]