摘要
目的探讨四逆散对皮质酮诱导PC12细胞损伤的干预作用是否与钙敏感受体(CaSR)以及细胞凋亡密切相关。方法以四逆散含药血清为干预药物,将PC12细胞分为空白血清组、皮质酮组、2.5%、5%、10%含药血清组,MTT法检测细胞存活率。通过AnnexinⅤ/PI流式细胞分析法、蛋白免疫印迹技术对细胞凋亡率以及C-Caspase3、C-Caspase9、Bax(6A7)、细胞Cytochrome C蛋白进行检测。通过AnnexinⅤ/PI流式细胞分析法检测四逆散含药血清联合CaSR激动剂(GdCl3)或CaSR抑制剂(NPS2390)各组细胞的凋亡率。结果与空白血清组比较,皮质酮组存活率、CaSR蛋白表达量降低(P<0.01,P<0.05);皮质酮组凋亡率、C-Caspase3、C-Caspase9、Bax(6A7)以及胞浆Cytochrome C蛋白表达量增加(P<0.01,P<0.05)。与皮质酮组比较,含药血清组存活率增加(P<0.01),5%、10%含药血清组CaSR蛋白表达量及皮质酮+NPS2390组凋亡率增加(P<0.01,P<0.05);含药血清组、皮质酮+GdCl3组凋亡率降低(P<0.01)和10%含药血清组C-Caspase3、C-Caspase9、Bax(6A7)、胞浆Cytochrome C蛋白表达量降低(P<0.01)。与皮质酮+10%含药血清组比较,皮质酮+10%含药血清+NPS2390组凋亡率增加(P<0.01)。与皮质酮+GdCl3组比较,皮质酮+10%含药血清+GdCl3组凋亡率差异无统计学意义(P>0.05)。结论四逆散含药血清可以抑制皮质酮诱导的PC12细胞凋亡,其作用机制与CaSR有关。
Objective To investigate whether the intervention effect of Sini San on corticosteroneinduced PC12 cell injury is closely related to calcium sensitive receptor(CaSR)and cells apoptosis.Methods With serum containing Sini San as the intervention drug,PC12 cells were divided into blank serum group,corticosterone group,2.5%,5%and 10%medicated serum group,and the cells survival rate was detected by MTT method.The cells apoptosis rate was determined using AnnexinⅤ/PI with flow cytometry.The expression of C-Caspase3,C-Caspase9,Bax(6 A7)and cytoplasmic Cytochrome C protein were detected by Western Blot.The apoptosis rate of each group cells in serum containing Sini San combined with CaSR agonist(GdCl3)or CaSR inhibitor(NPS2390)was determined by AnnexinⅤ/PI with flow cytometry.Results Compared with the blank serum group,the survival rate and CaSR protein expression in corticosterone group decreased(P<0.01,P<0.05).The apoptosis rate,the expression of C-Caspase3,C-Caspase9,Bax(6 A7)and cytoplasmic Cytochrome C protein increased in corticosterone group(P<0.01,P<0.05).Compared with the corticosterone group,the survival rate of medicated serum groups were increased(P<0.01),the CaSR protein expression of the 5%,10%medicated serum groups,and the apoptosis rate of corticosterone+NPS2390 group were increased(P<0.01,P<0.05);The apoptotic rate in medicated serum groups and corticosterone+GdCl3 decreased(P<0.01),the expression levels of C-Caspase3,C-Caspase9,Bax(6 A7)and cytoplasmic Cytochrome C protein were reduced in 10%serum group(P<0.01).Compared with corticosterone+10%medicated serum group,the apoptosis rate in corticosterone+10%medicated serum+NPS2390 group increased(P<0.01).There are no significant change in the apoptosis rate in corticosterone+10%medicated serum+GdCl3 group,compared with the corticosterone+GdCl3.Conclusion Sini San medicated serum can inhibit the apoptosis of PC12 cells induced by corticosterone,and its effect is related to the CaSR.
作者
莫海欣
张荣
王坤
沈崇坤
董钊扬
张劲
史亚飞
MO Hai-xin;ZHANG Rong;WANG Kun;SHEN Chong-kun;DONG Zhao-yang;ZHANG Jin;SHI Ya-fei(School of Basic Medicine Sciences,Guangzhou University of Chinese Medicine,Guangzhou,510006;Clinical Experimental Center,Jiangmen Central Hospital,Guangdong,529030;International Institute for Translational Chinese Medicine,Guangzhou University of Chinese Medicine,Guangzhou,510006;School of Nursing,Guangzhou University of Chinese Medicine,Guangzhou,510006)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2022年第2期187-194,共8页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目(No.81873271,No.82074219)
广东省自然科学基金资助项目(No.2021A1515012572)
广州市科技计划项目(No.202002030009)
广州中医药大学科研团队项目(No.2019KYTD301)。
关键词
PC12细胞
四逆散
抑郁症
凋亡
钙敏感受体
PC12 cells
Sini San
depression
apoptosis
calcium sensitive receptor
