糖络宁减轻大鼠背根神经节细胞炎症反应的作用机制研究

Mechanism of Tangluoning in Alleviating Inflammation of Dorsal Root Ganglion Cells in Rats

目的:以施万细胞外泌体为切入点,观察糖络宁含药血清对体外高糖诱导的大鼠背根神经节细胞炎症反应的影响。方法:采用糖络宁含药血清干预高糖培养的施万细胞,分为正常组、高糖组、糖络宁组,干预48 h后,提取各组细胞上清液中外泌体,将各组外泌体分别干预正常糖和高糖条件培养的大鼠背根神经节细胞,分为无外泌体-正常糖组,正常外泌体-正常糖组、高糖外泌体-正常糖组、糖络宁外泌体-正常糖组、无外泌体-高糖组、正常外泌体高糖组、高糖外泌体-高糖组、糖络宁外泌体-高糖组8个组,48 h后,收集各组细胞并进行相关检测。CCK-8法检测各组细胞活力,酶联免疫法检测各组细胞上清液中炎症介质(TNF-α、IL-6、IL-1β)的蛋白表达情况,实时荧光定量PCR检测各组细胞炎症介质的表达情况。结果:与正常外泌体-正常糖组比较,高糖外泌体-正常糖组细胞活力下降(P<0.05),炎症介质的基因和蛋白表达均增加(P<0.05);而与高糖外泌体-正常糖组比较,糖络宁外泌体-正常糖组细胞活力有所升高(P<0.05),各炎症介质的基因和蛋白表达均有所下调(P<0.05)。高糖条件培养的背根神经节细胞中,不同组外泌体干预表现出类似的作用效果;结论:糖络宁能通过调控施万细胞外泌体减轻大鼠背根神经节细胞炎症反应。

Objective:To observe the effect of Tangluoning-containing serum on inflammation of rat dorsal root ganglion cells exposed to high glucose in vitro and explore the mechanism of Tangluoning in protecting the dorsal root ganglion cells.Methods:The Schwann cells cultured with high glucose were classified into normal,high glucose,and Tangluoning groups.After 48 h of treatment,exosomes were extracted from the supernatant of cell culture.The exosomes of each group were used to intervene in the rat dorsal root ganglion cells cultured with and without high glucose,respectively.The cells were grouped as follows:no exosome+normal glucose,exosomes from normal cells+normal glucose,exosomes from cells cultured with high glucose+normal glucose,exosomes from cells cultured with Tangluoning+normal glucose,no exosome+high glucose,exosomes from normal cells+high glucose,exosomes from cells cultured with high glucose+high glucose,and exosomes from cells cultured with Tangluoning+high glucose.After 48 h,cells in each group were collected.The cell counting kit-8(CCK-8)assay was used to examine the cell viability in each group.Enzyme-linked immunosorbent assay was employed to measure the levels of inflammatory mediators[tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and IL-1β]in the supernatant of cell culture.Real-time fluorescence quantitative PCR was employed to determine the mRNA levels of TNF-α,IL-6,and IL-1β.Results:Compared with the group of exosomes from normal cells+normal glucose,the group of exosomes from cells cultured with high glucose+normal glucose showed decreased cell viability(P<0.05)and up-regulated expression of TNF-α,IL-6,and IL-1β(P<0.05).Compared with the group of exosomes from cells cultured with high glucose+normal glucose,the group of exosomes from cells cultured with Tangluoning+normal glucose showed increased cell viability(P<0.05)and down-regulated expression of inflammatory mediators(P<0.05).Different groups of exosomes showed similar effects on the dorsal root ganglion cells cultured with high glucose.Conclusion:Tangluoning can reduce the inflammation of dorsal root ganglion cells in rats by regulating the exosomes derived from Schwann cells.