文献类型：期刊文献

中文题名：富硒黄芪皂苷对IL-1β诱导大鼠退变软骨细胞mTOR信号通路的影响

英文题名：Effect of seleno enriched Astragalus Saponin on mTOR signal pathway in rat s degenerated chondrocytes induced by interleukin-1β

作者：颜春鲁[1];安方玉[2];金华[3];李嘉进[1];李孟翰[1];王国文[4];邓婕[4];张金龙[4];郭丹[4];彭霞[5]

第一作者：颜春鲁

机构：[1]甘肃中医药大学中医临床学院,兰州730000;[2]甘肃中医药大学教学实验实训中心,兰州730000;[3]甘肃中医药大学公共卫生学院,兰州730000;[4]甘肃中医药大学第一临床医学院,兰州730000;[5]甘肃中医药大学中西医结合学院,兰州730000

第一机构：甘肃中医药大学中医临床学院

年份：2021

卷号：14

期号：4

起止页码：382

中文期刊名：中华骨质疏松和骨矿盐疾病杂志

外文期刊名：Chinese Journal Of Osteoporosis And Bone Mineral Research

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：甘肃省自然科学基金(20JR5RA185,20JR5RA616);甘肃省高等学校创新能力提升项目(2019A-080);甘肃省高等学校科研基金(2018A-042);甘肃中医药大学科学研究与创新基金(2021KCYB-12)。

语种：中文

中文关键词：富硒黄芪皂苷;退变软骨细胞;mTOR信号通路

外文关键词：seleno enriched Astragalus Saponin;degenerated chondrocytes;mTOR signal pathway

摘要：目的探讨富硒黄芪皂苷对白细胞介素-1β(interleukin-1β,IL-1β)诱导的大鼠退变软骨细胞Ⅱ型胶原蛋白、磷酯酰肌醇-3-激酶(phosphatidylinositol 3 kinase,PI3K)、蛋白激酶B(protein kinase B,Akt)、哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)、Beclin-1和微管相关蛋白1轻链3(microtubulerassociated protein 1 light chain 3-Ⅱ,LC3-Ⅱ)mRNA表达的影响。方法体外退变软骨细胞模型的建立用IL-1β(10 ng/mL)诱导,模型组和富硒黄芪皂苷治疗组分别给予DEME培养基和富硒黄芪皂苷干预(25~300 mmol/L),另外将第3代培养48 h的软骨细胞作为正常对照组,给予DEME培养基干预24、48、72 h。应用噻唑蓝(MTT)法检测各组软骨细胞的增殖能力,运用免疫组化法观察各组软骨细胞Ⅱ型胶原蛋白的表达变化,运用RT-PCR法检测各组软骨细胞PI3K、Akt、mTOR、Beclin-1和LC3-Ⅱ的基因表达变化。结果正常对照组、模型组和100 mmol/L富硒黄芪皂苷治疗组的Ⅱ型胶原蛋白表达水平分别为2347.85±179.23,1547.27±261.79和3575.46±287.56;3组的PI3K基因表达水平分别为1.00±0.03,2.75±0.02和1.14±0.01;3组的Akt基因表达水平分别为1.00±0.04,1.83±0.07和1.13±0.04;3组的mTOR基因表达水平分别为1.00±0.02,2.12±0.04和1.52±0.03;3组的Beclin-1基因表达水平分别为1.02±0.02,0.61±0.03和0.90±0.02;3组的LC3-Ⅱ基因表达水平分别为0.97±0.02,1.24±0.01和1.05±0.04。与正常对照组比较,模型组退变软骨细胞的增殖能力在24、48、72 h均减弱,且其Ⅱ型胶原蛋白的表达和Beclin-1基因表达在48 h下降,而模型组退变软骨细胞PI3K、Akt和mTOR基因表达在48 h则增强(P<0.05)。与模型组比较,富硒黄芪治疗组退变软骨细胞增殖能力在24、48、72 h均增强,但以48 h的100 mmol/L富硒黄芪皂苷的差异最显著,同时其Ⅱ型胶原蛋白的表达和Beclin-1基因表达也升高,而100 mmol/L富硒黄芪则使PI3K、Akt和mTOR基因表达降低(P<0.05)。结论富硒黄芪皂苷延缓及控制骨关节炎进展的可能机制与调控mTOR信号通路有关。
Objective To observe the effects of seleno enriched Astragalus Saponin on the mRNA expression of collagen-Ⅱ,phosphatidylinositol 3 kinase(PI3K),protein kinase B(Akt),mammalian target of rapamycin(mTOR),Beclin-1,and microtubulerassociated protein 1 light chain 3-Ⅱ(LC3-Ⅱ)in rat s degenerated chondrocytes induced by interleukin-1β(IL-1β).Methods The degenerated chondrocytes models in vitro were established by IL-1β(10 ng/mL),which were intervened by the DEME medium(the model group)and seleno enriched Astragalus Saponin(25-300 mmol/L,the seleno enriched Astragalus Saponin group)and the third generation of 48 h chondrocytes was cultured by the normal control group,which was intervened by the DEME medium for 24,48,and 72 h.The MTT method was used to observe the cell proliferation of degenerated chondrocytes in each group.The immune-histochemistry method was used to observe the expression of degenerated chondrocytes collagen-Ⅱin each group.The RT-PCR method was used to detect the expression of degenerated chondrocytes PI3K,Akt,mTOR,Beclin-1,and LC3-ⅡmRNA in each group.Results The collagen typeⅡprotein in normal control group,model group,100 mmol/L seleno enriched Astragalus Saponin intervention group were 2347.85±179.23,1547.27±261.79,3575.46±287.56.The PI3K gene expre-ssion in the three groups were 1.00±0.03,2.75±0.02,1.14±0.01.The Akt gene expression in the three groups were 1.00±0.04,1.83±0.07,1.13±0.04.The mTOR gene expression in the three groups were 1.00±0.02,2.12±0.04,1.52±0.03.The Beclin-1 gene expression in the three groups were 1.02±0.02,0.61±0.03,0.90±0.02.The LC3-Ⅱgene expression in the three groups were 0.97±0.02,1.24±0.01,1.05±0.04.Compared with normal control group,the cell proliferation ability of degenerated chondrocytes of model group was dropped significantly in 24,48,and 72 h,and expression of collagen-Ⅱand Beclin-1 in degenerated chondrocytes was decreased significantly in 48 h,while the gene expressions of PI3K,Akt,and mTOR of degenerated chondrocytes were significantly increased in 48 h(P<0.05).Compared with model group,the cell proliferation ability of degenerated chondrocytes in seleno enriched Astragalus Saponin intervention group was significantly increased in 24,48,and 72 h,but only 100 mmol/L seleno enriched Astragalus Saponin intervention group had significant difference in 48 h,and expression of collagen-Ⅱand Beclin-1 in degenerated chondrocytes was significantly increased,while the gene expressions of PI3K,Akt,and mTOR of degenerated chondrocytes were significantly decreased(P<0.05).Conclusion The possible mechanism of seleno enriched Astragalus Saponin is related to the regulation of mTOR signaling pathway,which is seleno enriched Astragalus Saponin functions for delaying and controlling the progression of OA.