文献类型：期刊文献

中文题名：红芪多糖对高糖培养雪旺细胞IL-1β/NF-κB信号通路的影响

英文题名：Effect of Hedysarum Polysaccharides on IL-1β/NF-κB Signal Pathway of Schwann Cells Cultured in High Glucose

作者：何流[1];金智生[2];陈彦旭[2];徐长青[2];张磊[2];李圆[2]

第一作者：何流

机构：[1]贵州中医药大学第二附属医院,贵州贵阳550000;[2]甘肃中医药大学,甘肃兰州730000

第一机构：贵州中医药大学第二附属医院,贵州贵阳550000

年份：2022

卷号：33

期号：8

起止页码：1801

中文期刊名：时珍国医国药

外文期刊名：Lishizhen Medicine and Materia Medica Research

收录：北大核心:【北大核心2020】；CSCD:【CSCD_E2021_2022】；

基金：国家自然科学基金(81660777)。

语种：中文

中文关键词：糖尿病周围神经病变;雪旺细胞;炎症反应;凋亡反应;NF-κB;IL-1β;Cleaved caspase-3

外文关键词：Diabetic peripheral neuropathy;Schwann cells;Inflammatory reaction;Apoptotic response;NF-κB;IL-1β;Cleaved caspase-3

摘要：目的通过观察红芪多糖(Hedysarum Polysacchcrides,HPS)对高糖培养下雪旺细胞(Schwann Cells,SCs)核转录因子-κB(nuclear factor kappa-B,NF-κB)、白细胞介素-1β(IL-1β)、切割型半胱氨酸天冬氨酸蛋白水解酶-3(Cleaved caspase-3)信号通路的影响,探讨红芪多糖在治疗糖尿病周围神经病变(DPN)过程中可能作用机制。方法选取出生5~7 d的SD乳鼠4只,从坐骨神经部位提取雪旺细胞,常规培养后,CCK8法筛高糖、HPS、α-LA实验浓度,随机分为正常组、高糖组、红芪多糖组、α-硫辛酸(α-LA)组,免疫蛋白印迹法(Western-blot)、逆转录-免疫酶联反应(RT-PCR)法检测NF-κB、IL-1β、Cleaved caspase-3蛋白与mRNA表达情况,免疫荧光法检测特异性蛋白S-100β及GFAP表达,Annexin V/PI双染法检测各组雪旺细胞的凋亡率。结果Western-blotting、RT-PCR结果显示,与正常组比较,高糖组NF-κB、IL-1β、Cleaved Caspase-3蛋白与mRNA表达显著升高,差异有统计学意义(P<0.01);与高糖组比较,HPS组、α-LA组NF-κB、IL-1β、Cleaved caspase-3蛋白与mRNA表达显著降低,差异有统计学意义(P<0.01)。免疫荧光法检测结果显示,雪旺细胞质中S-100β和GFAP染色呈绿色荧光表达,而阳性对照组只见DAPI核染,未见绿色荧光染色。Annexin V/PI双染法检测结果显示,与正常组比较,高糖组雪旺细胞凋亡率明显增加;与高糖组比较,HPS组和α-LA组雪旺细胞凋亡率显著降低,差异有统计学意义(P<0.01)。结论红芪多糖可以减轻雪旺细胞炎症状态下的凋亡反应,其作用机制可能与抑制IL-1β/NF-κB信号通路激活有关。
Objective To observe the effects of Hedysarum Polysacchcrides(HPS)on Schwann Cells(SCs)nuclear factor kappa-B(NF-κB),interleukin-1β(IL-1β)and cleaved cysteinyl aspartate-specific protease-3(Cleaved caspase-3)signaling pathway in high glucose culture,and explore the possible mechanism of Hedysarum Polysacchcrides in the treatment of diabetic peripheral neuropathy(DPN).Methods Schwann cells were extracted from the sciatic nerve of four SD sucklimg rats aged 5 to 7 days.After routine culture,the suitable concentrations of high glucose,HPS andα-LA were screened by CCK8 method,and the cells were randomly divided in to nomal group,high hlucose group,HPS group andα-LA group.The protein and mRNA expres?sions of NF-κB,IL-1βand Cleaved caspase-3 were detected by Western-blot and RT-PCR.Immunofluorescence staining was used to detect the expression of specific protein Smuri 100βand GFAP,and AnnexinV/PI flow cytometry was used to detect the apoptosis rate of Schwann cells in each group.Results The results of Western-blotting and RT-PCR showed that the protein and mRNA expression levels of NF-κB,IL-1βand Cleaved Caspase-3 in high glucose group were significantly higher than those in normal group,while the expression levels of NF-κB,IL-1βand Cleaved caspase-3 in HPS group andα-LA group were significantly lower than those in high glucose group(P<0.01).The results of immunofluorescence staining showed that Smur100βand GFAP staining were green fluorescent staining in Schwann cytoplasm,while only DAPI nuclear staining was found in the positive control group,but no green fluorescent staining was found in the positive control group.The results of AnnexinV/PI flow cytometry showed that the apoptosis rate of Schwann cells in high glucose group was significantly higher than that in normal group,while that in HPS group andα-LA group was significantly lower than that in high glucose group(P<0.01).Conclusion Hedysarum polysaccharides can reduce the apoptosis of Schwann cells in inflammatory state,and its mechanism may be related to the inhibition of the activation of IL-1β/NF-κB signal pathway.