Effects of Tongluoxingnao effervescent tablet on Aβmetabolism in transgenic cell model
10.3969/j.issn.1001-1978.2016.11.018
- VernacularTitle:基于转基因细胞模型研究通络醒脑泡腾片对Aβ代谢的影响
- Author:
Wenjun FU
;
Yuan DAI
;
Jiangping WEI
;
Hang ZHENG
;
Tao MA
;
Shijun XU
;
Yongyan WANG
- Publication Type:Journal Article
- Keywords:
Alzheimer ’ s disease;
Tongluoxingnao ef-fervescent tablet;
SH-SY5 Y-APP cells;
SH-SY5 Y-C99 cells;
Aβ generation;
Aβ degradation
- From:
Chinese Pharmacological Bulletin
2016;32(11):1571-1577,1578
- CountryChina
- Language:Chinese
-
Abstract:
Aim Tongluoxingnao effervescent tablets ( TLXNET) ,based on the ancient formula of QiongGui Tang, can improve cognitive dysfunction in different AD models. This research is aimed to study the effects of TLXNET on the Aβmetabolism and explore the anti-AD mechanism in SH-SY5 Y-APP and SH-SY5 Y-C99 cells. Methods Cells were incubated for 48h in dif-ferent concentrations of medicated serum ( containing 0% ~40% of TLXNET in the serum ) . Firstly, the non-toxic concentration was measured by MTT assay, the activity of cells was detected by LDH methods, and ELISA was used to measure the levels of Aβ1-40 and Aβ1-42 . Then, the gene and protein expressions of APP were detected via RT-PCR and Western blot of which the expressions of shearing fragments such as sAPPα and sAPPβ were investigated by Western blot, the same as the expressions of IDE and NEP. Addi-tionally, the level of mRNA, protein and activity of BACE1 were measured by qRT-PCR, Western blot and fluorescence detection kits respectively. Eventually, MTT assay was performed to detect the cell viability after the SH - SY 5 Y cells were treated with various concentrations of medicated serum and Aβ25-35 for 48h. Results There was no significant toxicity of TLXNET medicated serum in SH-SY5 Y-APP and SH-SY5 Y-C99 cells ( P >0 . 05 ) . TLXNET could signifi-cantly inhibit Aβ secretion in SH-SY5 Y-APP cells ( P<0 . 05 ) , but had no effect on Aβ secretion in SH-SY5Y-C99 cells, the same as the level of APP mRNA and protein in SH-SY5 Y-APP cells and the expressions of IDE and NEP (P>0. 05). Additionally, TLXNET could still notably inhibit the expression of sAPPβ pro-tein in a dose-dependent way, with statistical signifi-cance ( P <0. 05 ) . Meanwhile, the level of mRNA, protein and activity of BACE1 were also significantly decreased by TLXNET (P <0. 01). Moreover, the medicated serum of TLXNET had a protective effect on SH-SY5 Y apoptosis induced by Aβ25-35 ( P <0 . 01 ) . Conclusion TLXNET could obviously inhibit β-secretase enzyme, and has an antagonistic effect a-gainst Aβneurotoxicity, which suggests that the inhibi-tion of β-secretase enzyme and antagonism against Aβneurotoxicity are the main anti-AD mechanism of TLX-NET .
