Effect of Huannao Yicong Decoction extract on activity ofγsecretase and γsecretase-related regulation pathway of human APP/PS 1 double transgenic cell line
10.3969/j.issn.1001-1978.2017.03.024
- VernacularTitle:还脑益聪方提取物含药血清对APP/PS 1双基因转染细胞γ分泌酶活性及相关蛋白表达的影响
- Author:
Yun WEI
;
Meixia LIU
;
Xiaodong LIANG
;
Jiangang LIU
;
Hao LI
;
Yun WEI
- Keywords:
Alzheimer's disease (AD);
γsecretase;
extract of Huannao Yicong Decoction;
serum containing drug;
human APP/PS1 double transgenic cell line;
Chi-nese medicine compound
- From:
Chinese Pharmacological Bulletin
2017;33(3):417-426
- CountryChina
- Language:Chinese
-
Abstract:
Aim Toobservetheeffectofconcisepre-scriptions of Chinese medicine Huannao Yicong Decoc-tion(HYD)on regulatory pathway of secretase in APP/PS1 double transgenic cell line(HEK293),and to in-vestgateitsmechanism.Methods Theproliferationof AD cell model and the toxicity of each investigational drugs were ebserved by CCK-8;the changes in micro-scopic structure of each group were observed by(Trans-mission electron microscope,TEM);the activities of gamma-secretes was observed by Dual Luciferase Re-porter Gene Assay Kit ,and then the expression of pre-senilin 1(PS1),carboxyl terminus of Hsc70-interacting protein(CHIP),GTP binding protein (CDC42 ),ante-rior pharynx defective-1α(APH-1α),Hypoxia induc-ible factor-1α(HIF-1α) were detected by Western blot.Results 15%HYDserumincreasedthecellac-tivity compared to blank serum (P <0. 01 ).Under TEM,human APP/PS1 Double Transgenic cell line's mitochondria showed swelling and degenerat,and lipo-fuscin was deposited;after 48 h of medication,theswelling of mitochondria was reduced and crista was clear,and the number of lipofuscin was reduced.For the activity ofγsecretase,after 24 h medication,com-pared to control group,HYD directly group and DAPT group obviously inhibited the activity ofγsecretase (P<0. 05 or P<0. 01 ),compared to DAPT group,only 15% serum containing HYD group had significant difference (P <0. 05 );after 48 h medication,com-pared to control group,the other group all showed sig-nificant difference(P<0. 05 or P<0. 01 ).For the in-fluence of PS pathway,after 48 h medication,com-pared to control group,the other group all inhibited the PS1protein expression(P<0. 05 or P<0. 01),HYD directly group could activate the CDC42 protein expres-sion(P<0. 05 );compared to 0 h midicaiton,DAPT group inhibited the CHIP protein expression at the point of 48 h(P<0. 05 )and activated the CDC42 pro-tein expression at the point of 24 h.For the influence of Aph-1 pathway,there was no significant difference for Aph-1α(P>0. 05 );compared to control group,HYD directly group inhibited the HIF-1αprotein ex-pression after 48h medication(P<0. 05);compared to 0h midicaiton,DAPT group inhibited the HIF-1αpro-tein expression at the point of 24 h (P<0. 05 ).Con-clusions HYDcantreatADthroughprotectingthe mitochondrial function,reducing the formation of lipo-fuscin,inhibiting the activity of γsecretase by down-regulating the activity of HIF-1α,decreasing the stabil-ity and activity of PS1 by promoting the expression of CDC42.This shows that HYD has good research and development prospect as an effective drug for preven-tion and treatment of AD.
