摘要
目的:探讨银杏黄酮苷元(GA)对雨蛙素诱导的急性胰腺炎(AP)大鼠腺泡细胞炎症因子分泌和凋亡的影响。方法:采用雨蛙素诱导大鼠胰腺腺泡细胞株AR42J建立AP腺泡细胞模型。①正常培养的AR42J细胞作为空白对照组,AP模型细胞为AP组,以用15、30、60 mg/L GA处理的AP模型细胞作为GA低、中、高浓度组。②将pcDNA、pcDNA-双特异性磷酸酶1(DUSP1)转染至AR42J细胞6 h后,再用100 nmol/L雨蛙素刺激,分别作为AP+pcDNA组、AP+pcDNA-DUSP1组。③将si-NC、si-DUSP1转染至AR42J细胞6 h后,再用100 nmol/L雨蛙素和60 mg/L的GA处理,分别作为AP+GA+si-NC组、AP+GA+si-DUSP1组,另取AP模型细胞作为AP组、用100 nmol/L雨蛙素和60 mg/L的GA处理的细胞作为AP+GA组。ELISA法检测培养上清中炎症因子IL-6、TNF-α水平;Annexin V-FITC/PI双染法检测细胞凋亡;Western blot法检测蛋白表达;qRT-PCR法检测DUSP1 mRNA的表达水平。结果:GA处理后,雨蛙素诱导的AR42J细胞中IL-6、TNF-α水平降低,细胞凋亡率升高,Bcl-2表达水平降低,Bax表达水平升高,DUSP1表达水平升高(P<0.05)。过表达DUSP1可降低雨蛙素诱导的AR42J细胞炎症因子水平,促进细胞凋亡(P<0.05)。干扰DUSP1表达逆转了GA对雨蛙素诱导的AP腺泡细胞炎症因子和凋亡的影响(P<0.05)。结论:GA可通过上调DUSP1表达抑制雨蛙素诱导的AP腺泡细胞炎症反应,促进细胞凋亡。
Aim:To investigate the effects of ginkgo flavone aglycone(GA)on secretion of inflammatory factors and apoptosis of caerulein-induced acute pancreatitis(AP)acinar cells.Methods:AP model was established by inducing AR42J cells with caerulein.Normal cultured cells were used as blank control group,AP model cells as AP group,AP model cells treated with 15,30 and 60 mg/L GA as GA low,medium and high concentration groups.pcDNA and pcDNA-dual specificity phosphatase 1(DUSP1)were transfected into AR42J cells for 6 hours,then stimulated by 100 nmol/L caerulein as AP+pcDNA and AP+pcDNA-DUSP1 group.si-NC,si-DUSP1 were transfected into AR42J cells for 6 hours,then stimulated by caerulein and GA(60 mg/L)as AP+GA+si-NC group and AP+GA+si-DUSP1 group;in addition,AP model cells were used as AP group,and cells treated with cerulein and GA were used as AP+GA group.The levels of IL-6 and TNF-αsupernatant were detected by ELISA;cell apoptosis was detected by Annexin V-FITC/PI double staining;the protein expressions were detected by Western blot;DUSP1 mRNA expression level was detected by qRT-PCR.Results:After GA treatment,the levels of IL-6 and TNF-αin the caerulein-induced AR42J cells were significantly reduced,the apoptosis rate was significantly increased,the expression of Bcl-2 was significantly reduced,the expression of Bax was significantly increased,and the expression of DUSP1 was significantly increased(P<0.05).Overexpression of DUSP1 could reduce the levels of inflammatory factors in caerulein-induced AR42J cells and promote cell apoptosis(P<0.05).Interference with DUSP1 expression reversed the effect of GA on the inflammatory factors and apoptosis of caerulein-induced acute AR42J cells(P<0.05).Conclusions:GA could inhibit AP acinar cells′inflammation reaction and promote apoptosis by up-regulating the expression of DUSP1.
作者
杜健磊
王俊苹
刘伟
DU Jianlei;WANG Junping;LIU Wei(Department of Pharmacy,Zhumadian Central Hospital,Zhumadian,Henan 463000;Department of Clinical Pharmacy,the Third Affiliated Hospital,Xinxiang Medical College,Xinxiang,Henan 453003)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2021年第2期270-274,共5页
Journal of Zhengzhou University(Medical Sciences)
基金
河南省重点科技攻关计划项目(172106310062)。
关键词
银杏黄酮苷元
双特异性磷酸酶1
急性胰腺炎
凋亡
大鼠
ginkgo flavone aglycone
dual specificity phosphatase 1
acute pancreatitis
apoptosis
rat
