Two isomers of HDTIC isolated from Astragali Radix decrease the expression of p16 in 2BS cells
Two isomers of HDTIC isolated from Astragali Radix decrease the expression of p16 in 2BS cells
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DOI：
10.1097/00029330-200802010-00009
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发表时间：
2008-02-05
影响因子：
6.1
通讯作者：
Tong Tan-Jun
中科院分区：
文献类型：
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作者：
Wang Pei-Chang;Zhang Zong-Yu;Tong Tan-Jun
背景黄芪（Astragalus membranceus（Fish）Bunge Var.）蒙古黄芪（Bge）是一种有效的中药抗衰老物质。4-羟基-5-羟甲基-[1，3]二氧戊环-2，6 '-螺环-5'，6 '，7'，8 '-四氢中氮茚-3'-甲醛（HDTIC）的两种异构体HDTIC-1和HDTIC-2于2002年首次从该草药中提取。我们以前证明，0.1 μ mol/L HDTIC-1或1.0 μ mol/L HDTIC-2强烈延迟人胎肺二倍体成纤维细胞（2BS）的复制衰老。本研究选择HDTIC-1和HDTIC-2作为研究对象，通过观察它们对衰老相关基因表达的影响，探讨HDTIC延缓复制性衰老的机制。方法采用RT-PCR和Western blot方法，观察HDTIC-1和HDTIC-2对p16和p21表达的影响。抗氧化剂处理后，通过表型改变观察化合物的抗氧化活性。结果p16蛋白在对照衰老细胞中有明显表达。而在2BS细胞中，在0.1 μ mol/L HDTIC-1或1.0 μ mol/L HDTIC-2中培养56个群体doubles（PD）后，观察到p16的弱mRNA表达和p16的蛋白表达。p2 l的表达水平随着细胞的老化而增加。此外，在对照细胞和用HDTIC化合物培养的相同PD细胞中的p21表达水平之间没有差异。结果还表明，暴露于100 μ mol/L H2 O2 5分钟的2BS细胞恢复其非衰老表型，并且在将受损细胞与HDTIC-1孵育后继续融合（1.0 μ mol/L）或HDTIC-2结论HDTIC化合物对2BS细胞p16表达有明显的抑制作用，这可能通过p16（INK 4a）/Rb/MAPK途径参与延缓细胞复制性衰老。HDTIC-1和HDTIC-2的抗氧化活性，在这项研究中首次被描述，可能是间接相关的抑制p16的表达。
Background Astragali Radix, the root of Astragalus membranceus (Fish) Bunge Var. mongholicus (Bge), is a crude drug considered as one of the effective traditional Chinese anti-ageing material. The two isomers of 4-hydroxy-5-hydroxymethyl-[1,3]dioxolan-2,6'-spirane-5',6',7',8'-tetrahydro-indolizine-3'-carbaldehyde (HDTIC), HDTIC-1 and HDTIC-2, were first extracted from the herb in 2002. We demonstrated previously that 0.1 mu mol/L HDTIC-1 or 1.0 mu mol/L HDTIC-2 strongly delay replicative senescence of human fetal lung diploid fibroblasts (2BS). In this study, we chose them to investigate their effects on the expression of senescence-associated genes to explore the mechanism of how HDTIC delays replicative senescence.Methods The effects of HDTIC-1 and HDTIC-2 on the expression of p16 and p21 were observed in vitro by RT-PCR and Western blot. The anti-oxidative activities of the compounds were also observed by phenotype alteration after treatment with antioxidants. Results There was an obvious expression of p16 in the control senescent cells. However, in the 2BS cells, after 56 population doublings (PDs) grown from PD28 in 0.1 mu mol/L HDTIC-1 or 1.0 mu mol/L HDTIC-2, there was a weak mRNA expression of p16 and no protein expression of p16 was observed. The expression level of p2l increased with cell ageing. Moreover, there was no difference between the expression level of p21 in the control cells and that in the same PD cells cultured with HDTIC compounds. The results also showed that 2BS cells exposed to 100 mu mol/L H2O2 for 5 minutes return to their non-senescent phenotype and continue to be confluent after incubating the damaged cells with HDTIC-1 (1.0 mu mol/L) or HDTIC-2 (10 mu mol/L) for 1 hour.Conclusions Expression of p16 by 2BS cells was strongly inhibited by HDTIC compounds, which could contribute to their delayed replicative senescence by the way of p16(INK4a)/Rb/MAPK. The anti-oxidative activities of HDTIC-1 and HDTIC-2, described in this study for the first time, might be indirectly related to their inhibition of p16 expression.