双丹明目胶囊含药血清对高糖诱导的大鼠视网膜神经节细胞损伤的改善作用及其机制OA
Alleviating effect of medicated serum of Shuangdan Mingmu Capsules on high glucose-induced injury in rat retinal ganglion cells and its mechanism
目的 探讨双丹明目胶囊含药血清对高糖诱导的大鼠视网膜神经节细胞(RGCs)损伤的改善作用及其机制.方法 将体外培养的大鼠RGCs分为空白血清对照组(5.5 mmol·L-1葡萄糖+体积分数10％空白血清)、含药血清对照组(5.5 mmol·L-1葡萄糖+体积分数10％双丹明目胶囊含药血清)、空白血清高糖组(35.0 mmol·L-1葡萄糖+体积分数10％空白血清)和含药血清治疗组(35.0 mmol·L-1葡萄糖+体积分数10％双丹明目胶囊含药血清).干预48 h后,采用CCK-8法检测细胞存活率,流式细胞术检测细胞凋亡率,Western blot检测RGCs中内质网应激相关蛋白——活化转录因子6、蛋白激酶RNA样内质网激酶(PERK)、C/EBP同源蛋白(CHOP)及真核翻译起始因子2α(eIF2α)磷酸化水平.结果 与空白血清对照组及含药血清对照组相比,空白血清高糖组细胞存活率显著降低,细胞凋亡率显著升高(均为P＜0.01).与空白血清高糖组相比,含药血清治疗组细胞存活率显著上升,细胞凋亡率显著下降(均为P＜0.01).与空白血清对照组及含药血清对照组相比,空白血清高糖组RGCs中活化转录因子6、PERK、CHOP及p-eIF2α蛋白表达水平均显著升高(均为P＜0.05).与空白血清高糖组相比,含药血清治疗组上述各蛋白的表达水平均显著下降(均为P＜0.05);各组eIF2α蛋白表达水平差异无统计学意义(P＞0.05).结论 双丹明目胶囊含药血清可有效减轻高糖诱导的大鼠RGCs损伤与凋亡,其机制与调控PERK/CHOP/eIF2α信号通路、降低细胞内质网应激水平密切相关.
Objective To investigate the mechanism by which the medicated serum of Shuangdan Mingmu Capsules alleviates high glucose-induced injury in rat retinal ganglion cells(RGCs).Methods Rat RGCs cultured in vitro were di-vided into the blank serum control group(5.5 mmol·L-1 glucose+10％volume fraction of blank serum),medicated serum control group(5.5 mmol·L-1 glucose+10％volume fraction of medicated serum of Shuangdan Mingmu Capsules),blank serum high glucose group(35.0 mmol·L-1 glucose+10％volume fraction of blank serum),and medicated serum treat-ment group(35.0 mmol·L-1 glucose+10％volume fraction of medicated serum of Shuangdan Mingmu Capsules).After 48 hours of intervention,cell viability was assessed using the CCK-8 assay,and the apoptosis rate was measured using flow cytometry.Western blot was used to detect the expression levels of endoplasmic reticulum stress-related proteins,including activating transcription factor 6(ATF6),protein kinase RNA-like endoplasmic reticulum kinase(PERK),C/EBP homolo-gous protein(CHOP),and the phosphorylation level of the eukaryotic translation initiation factor 2 alpha(eIF2α).Results Compared with the blank serum control group and the medicated serum control group,the blank serum high glucose group exhibited a significantly decreased cell viability and a significantly increased apoptosis rate(all P＜0.01).Compared with the blank serum high glucose group,the medicated serum treatment group showed a significantly increased cell viabili-ty and a significantly decreased apoptosis rate(both P＜0.01).Compared with the blank serum control group and the medi-cated serum control group,the expression levels of ATF6,PERK,CHOP,and p-eIF2α in RGCs significantly increased in the blank serum high glucose group(all P＜0.05).Compared with the blank serum high glucose group,the expression levels of the above proteins significantly decreased in the medicated serum treatment group(all P＜0.05).No statistically significant difference was observed in the expression level of eIF2α among the groups(all P＞0.05).Conclusion The medicated serum of Shuangdan Mingmu Capsules significantly alleviates high glucose-induced RGC damage and apoptosis in rats,which may be closely related to its role in regulating the PERK/CHOP/eIF2α signaling pathway and reducing the level of en-doplasmic reticulum stress in cells.
刘峥嵘;杨婷;赵永旺;秦裕辉
410006 湖南省长沙市,湖南省中西医结合医院(湖南省中医药研究院附属医院)五官科410006 湖南省长沙市,湖南省中西医结合医院(湖南省中医药研究院附属医院)五官科201699 上海市,上海交通大学医学院附属松江医院眼科410006 湖南省长沙市,湖南省中西医结合医院(湖南省中医药研究院附属医院)五官科
医药卫生
双丹明目胶囊糖尿病视网膜病变内质网应激细胞凋亡
Shuangdan Mingmu Capsulesdiabetic retinopathyendoplasmic reticulum stressapoptosis
《眼科新进展》 2026 (6)
432-436,5
国家自然科学基金项目(编号:81874493)湖南省自然科学基金项目(编号:2022JJ40237)
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