人参皂甙Rd降低Nod样受体蛋白3表达保护神经细胞创伤性损伤的作用

Ginsenoside Rd Inhibits Nod-like Receptor Protein 3 Expression and Protects Neural Cells from Pyroptosis with Traumatic Brain Injury

目的探讨人参皂甙Rd(GSRd)治疗Nod样受体蛋白3(NLRP3)保护神经细胞创伤性损伤的作用和分子机制。方法培养HT22细胞系,随机分为空白组(Sham组)、生理盐水组(Con组)和GSRd治疗组(GSRd组),GSRd组又分为10μmol/L和50μmol/L两个亚组。先对GSRd组和Con组进行划伤6 h制成神经细胞创伤性脑损伤模型,再恢复正常培养液培养后加入GSRd培养48 h,最后取材。采用细胞增殖-毒性检测(CCK8法)检测细胞活力。利用荧光免疫组化法检测NLRP3的蛋白水平;利用蛋白质印迹技术检测NLRP3的表达变化。酶联免疫吸附测定检测白细胞介素-1β(IL-1β)和白细胞介素-18(IL-18)表达变化。结果Con组的细胞活力明显低于Sham组,不同剂量GSRd组的细胞活力高于Con组(P<0.05)。与Sham组比较,Con组HT22细胞中NLRP3的染色阳性率高;与Con组比较,GSRd组的NLRP3表达明显降低(P<0.05),且IL-1β和IL-18表达也明显减少(P<0.05)。结论GSRd可明显下调NLRP3表达水平,并减轻IL-1β和IL-18等炎性因子的释放,发挥治疗创伤性脑损伤的重要作用。

Objective To investigate the roll of ginsenoside Rd(GSRd)in regulating the expression of inflammatory related protein Nod-like receptor protein 3(NLRP3)and protecting neural cells from pyroptosis with traumatic brain injury and its molecular mechanism.Methods HT22 cell line was cultured and randomly divided into blank group(Sham group),normal saline control group(Con group)and GSRd treatment group(GSRd group).And the GSRd group was further divided into 10μmol/L and 50μmol/L subgroups.The GSRd groups and Con group were scratched for 6 h to make models of traumatic brain injury of neuronal cells.Then normal culture medium was restored and GSRd was added to culture for 48 h.And finally the samples were collected.Cell viability was detected by cell counting kit-8(CCK8)assay.The level of NLRP3 was detected by fluorescence immunohistochemistry.The changes of NLRP3 expression were detected by Western blotting.The changes of the expression of IL-1βand IL-18 was detected by standard enzyme linked immunosorbent assay(ELISA)kit.Results Cell viability of Con group was significantly lower than that of Sham group,and cell viability of GSRd groups with different dosages was higher than that of Con group(P<0.05).Compared with Sham group,the positive rate of NLRP3 staining in Con group was high.Compared with Con group,the expression of NLRP3,IL-1βand IL-18 in GSRd groups was significantly decreased(P<0.05).Conclusion GSRd can significantly down-regulate the expression level of NLRP3,and reduce the release of inflammatory factors such as IL-1βand IL-18,which plays an important role in the treatment of traumatic brain injury.