title: ÐÐ»Ð¸ÑÐ½Ð¸Ðµ Ð°Ð¼Ð±ÑÐ¾ÐºÑÐ¾Ð»Ð° Ð½Ð° ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½ÑÑ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ Ð³Ð»ÑÐºÐ¾ÑÐµÑÐµÐ±ÑÐ¾Ð·Ð¸Ð´Ð°Ð·Ñ Ð½Ð° Ð¿ÐµÑÐ²Ð¸ÑÐ½Ð¾Ð¹ ÐºÑÐ»ÑÑÑÑÐµ Ð¼Ð°ÐºÑÐ¾ÑÐ°Ð³Ð¾Ð²
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð±Ð°ÐºÐ°Ð»Ð°Ð²ÑÐ°
titleType: Subtitle
title: Ambroxol influence on the enzymatic glucocerebrosidase activity in primary macrophage culture
Creators: name: ÐÐ·ÑÐ¼ÑÐµÐ½ÐºÐ¾, ÐÑÑÐµÐ¼
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2020
Subjects: Ð±Ð¾Ð»ÐµÐ·Ð½Ñ ÐÐ°ÑÐºÐ¸Ð½ÑÐ¾Ð½Ð°
GBA
GCase
HexSph
Ð¼Ð°ÐºÑÐ¾ÑÐ°Ð³Ð¸ Ð¿ÐµÑÐ¸ÑÐµÑÐ¸ÑÐµÑÐºÐ¾Ð¹ ÐºÑÐ¾Ð²Ð¸
ÑÐ°ÑÐ¼Ð°ÐºÐ¾Ð»Ð¾Ð³Ð¸ÑÐµÑÐºÐ¸Ðµ ÑÐ°Ð¿ÐµÑÐ¾Ð½Ñ
Parkinson's disease
peripheral blood macrophages
pharmacological chaperones

description: ÐÐ¸Ð¿Ð»Ð¾Ð¼Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Â«ÐÐ»Ð¸ÑÐ½Ð¸Ðµ Ð°Ð¼Ð±ÑÐ¾ÐºÑÐ¾Ð»Ð° Ð½Ð° ÑÐµÑÐ¼ÐµÐ½ÑÐ°ÑÐ¸Ð²Ð½ÑÑ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ
Ð³Ð»ÑÐºÐ¾ÑÐµÑÐµÐ±ÑÐ¾Ð·Ð¸Ð´Ð°Ð·Ñ Ð½Ð° Ð¿ÐµÑÐ²Ð¸ÑÐ½Ð¾Ð¹ ÐºÑÐ»ÑÑÑÑÐµ Ð¼Ð°ÐºÑÐ¾ÑÐ°Ð³Ð¾Ð²Â» Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° Ð¾ÑÐµÐ½ÐºÐµ
Ð²Ð»Ð¸ÑÐ½Ð¸Ñ ÑÐ°ÑÐ¼Ð°ÐºÐ¾Ð»Ð¾Ð³Ð¸ÑÐµÑÐºÐ¾Ð³Ð¾ ÑÐ°Ð¿ÐµÑÐ¾Ð½Ð° (Ð¤Ð¨) Ð°Ð¼Ð±ÑÐ¾ÐºÑÐ¾Ð»Ð° Ð½Ð° Ð¸Ð·Ð¼ÐµÐ½ÐµÐ½Ð¸Ðµ
Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ ÑÐµÑÐ¼ÐµÐ½ÑÐ° Ð³Ð»ÑÐºÐ¾ÑÐµÑÐµÐ±ÑÐ¾Ð·Ð¸Ð´Ð°Ð·Ñ (GCase) Ð¸ ÑÑÐ¾Ð²Ð½Ñ
Ð»Ð¸Ð·Ð¾ÑÑÐ¸Ð½Ð³Ð¾Ð»Ð¸Ð¿Ð¸Ð´Ð° Ð³ÐµÐºÑÐ¾Ð·Ð¸Ð»ÑÑÐ¸Ð½Ð³Ð¾Ð»Ð¸Ð·Ð° (HexSph) Ð² ÑÑÑÐ¸Ñ Ð¿ÑÑÐ½Ð°Ñ
Ð¼Ð°ÐºÑÐ¾ÑÐ°Ð³Ð¾Ð² Ð¿ÐµÑÐ¸ÑÐµÑÐ¸ÑÐµÑÐºÐ¾Ð¹ ÐºÑÐ¾Ð²Ð¸ Ð¿Ð°ÑÐ¸ÐµÐ½ÑÐ¾Ð² Ñ Ð±Ð¾Ð»ÐµÐ·Ð½ÑÑ ÐÐ°ÑÐºÐ¸Ð½ÑÐ¾Ð½Ð°,
Ð°ÑÑÐ¾ÑÐ¸Ð¸ÑÐ¾Ð²Ð°Ð½Ð½Ð¾Ð¹ Ñ Ð¼ÑÑÐ°ÑÐ¸ÑÐ¼Ð¸ Ð² Ð³ÐµÐ½Ðµ GBA (GBA-ÐÐ). Ð ÑÐ¾Ð´Ðµ ÑÐ°Ð±Ð¾ÑÑ Ð±ÑÐ»Ð¾
Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð¾ ÐºÑÐ»ÑÑÐ¸Ð²Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ Ð¼Ð°ÐºÑÐ¾ÑÐ°Ð³Ð¾Ð² Ð¿ÐµÑÐ¸ÑÐµÑÐ¸ÑÐµÑÐºÐ¾Ð¹ ÐºÑÐ¾Ð²Ð¸ Ð¿Ð°ÑÐ¸ÐµÐ½ÑÐ¾Ð² Ñ
GBA-ÐÐ, Ð° ÑÐ°ÐºÐ¶Ðµ Ð½ÐµÐ²ÑÐ»Ð¾Ð³Ð¸ÑÐµÑÐºÐ¸ Ð·Ð´Ð¾ÑÐ¾Ð²ÑÑ Ð¸Ð½Ð´Ð¸Ð²Ð¸Ð´ÑÑÐ¼Ð¾Ð² (ÐºÐ¾Ð½ÑÑÐ¾Ð»Ñ) Ð²
Ð¿ÑÐ¸ÑÑÑÑÑÐ²Ð¸Ð¸ Ð¸ Ð±ÐµÐ· Ð°Ð¼Ð±ÑÐ¾ÐºÑÐ¾Ð»Ð°, Ñ Ð¿Ð¾ÑÐ»ÐµÐ´ÑÑÑÐ¸Ð¼ ÑÐ¾ÑÐ¼Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸ÐµÐ¼ Ð±Ð°Ð½ÐºÐ° ÑÑÑÐ¸Ñ
Ð¿ÑÑÐµÐ½ Ð¼Ð°ÐºÑÐ¾ÑÐ°Ð³Ð¾Ð² Ð²ÑÐµÑ Ð¸Ð½Ð´Ð¸Ð²Ð¸Ð´ÑÑÐ¼Ð¾Ð², Ð²Ð¾ÑÐµÐ´ÑÐ¸Ñ Ð² Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ðµ, Ð´Ð»Ñ
Ð´Ð°Ð»ÑÐ½ÐµÐ¹ÑÐµÐ¹ Ð¾ÑÐµÐ½ÐºÐ¸ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ GCase Ð¸ ÐºÐ¾Ð½ÑÐµÐ½ÑÑÐ°ÑÐ¸Ð¸ HexSph Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼
ÐÐ­ÐÐ¥ ÐÐ¡/ÐÐ¡.
ÐÐ¸Ð¿Ð»Ð¾Ð¼Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð²ÑÐ¿Ð¾Ð»Ð½ÐµÐ½Ð° Ð½Ð° Ð±Ð°Ð·Ðµ Ð»Ð°Ð±Ð¾ÑÐ°ÑÐ¾ÑÐ¸Ð¸ Ð¼Ð¾Ð»ÐµÐºÑÐ»ÑÑÐ½Ð¾Ð¹ Ð³ÐµÐ½ÐµÑÐ¸ÐºÐ¸
ÑÐµÐ»Ð¾Ð²ÐµÐºÐ° ÐÐÐ¦ Â«ÐÑÑÑÐ°ÑÐ¾Ð²ÑÐºÐ¸Ð¹ Ð¸Ð½ÑÑÐ¸ÑÑÑÂ» - ÐÐÐ¯Ð¤. Ð ÑÐ¾Ð´Ðµ Ð´Ð°Ð½Ð½Ð¾Ð¹ ÑÐ°Ð±Ð¾ÑÑ
Ð±ÑÐ»Ð¾ Ð¿Ð¾ÐºÐ°Ð·Ð°Ð½Ð¾, ÑÑÐ¾ Ð¼Ð°ÐºÑÐ¾ÑÐ°Ð³Ð¸ Ð¿ÐµÑÐ¸ÑÐµÑÐ¸ÑÐµÑÐºÐ¾Ð¹ ÐºÑÐ¾Ð²Ð¸ ÑÐµÐ»Ð¾Ð²ÐµÐºÐ° ÑÐ²Ð»ÑÑÑÑÑ
ÑÐ¾ÑÐ¾ÑÐµÐ¹ Ð¼Ð¾Ð´ÐµÐ»ÑÑ Ð´Ð»Ñ Ð¾ÑÐµÐ½ÐºÐ¸ Ð¸Ð·Ð¼ÐµÐ½ÐµÐ½Ð¸Ñ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ GCase Ð¸ ÐºÐ¾Ð½ÑÐµÐ½ÑÑÐ°ÑÐ¸Ð¸
ÐµÐµ ÑÑÐ±ÑÑÑÐ°ÑÐ° Ð¿ÑÐ¸ Ð²Ð¾Ð·Ð´ÐµÐ¹ÑÑÐ²Ð¸Ð¸ Ð½Ð° Ð½Ð¸Ñ Ð»ÐµÐºÐ°ÑÑÑÐ²ÐµÐ½Ð½ÑÐ¼Ð¸ Ð¿ÑÐµÐ¿Ð°ÑÐ°ÑÐ°Ð¼Ð¸. ÐÐ¿ÐµÑÐ²ÑÐµ
Ð¿Ð¾ÐºÐ°Ð·Ð°Ð½Ð¾, ÑÑÐ¾ Ð°Ð¼Ð±ÑÐ¾ÐºÑÐ¾Ð» ÑÑÑÐµÐºÑÐ¸Ð²Ð½Ð¾ Ð²Ð¾ÑÑÑÐ°Ð½Ð°Ð²Ð»Ð¸Ð²Ð°ÐµÑ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ GCase Ð¸
ÑÐ½Ð¸Ð¶Ð°ÐµÑ ÐºÐ¾Ð½ÑÐµÐ½ÑÑÐ°ÑÐ¸Ñ HexSph Ð² Ð¼Ð°ÐºÑÐ¾ÑÐ°Ð³Ð°Ñ Ð¿ÐµÑÐ¸ÑÐµÑÐ¸ÑÐµÑÐºÐ¾Ð¹ ÐºÑÐ¾Ð²Ð¸
Ð¿Ð°ÑÐ¸ÐµÐ½ÑÐ¾Ð² Ñ GBA-ÐÐ, ÑÑÐ¾ Ð¿Ð¾ÐºÐ°Ð·ÑÐ²Ð°ÐµÑ Ð¿Ð¾ÑÐµÐ½ÑÐ¸Ð°Ð»ÑÐ½ÑÑ Ð²Ð¾Ð·Ð¼Ð¾Ð¶Ð½Ð¾ÑÑÑ
Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð¸Ñ Ð´Ð°Ð½Ð½Ð¾Ð³Ð¾ Ð¤Ð¨ Ð² ÑÐµÑÐ°Ð¿Ð¸Ð¸ GBA-ÐÐ.
ÐÑÐ¸ Ð²ÑÐ¿Ð¾Ð»Ð½ÐµÐ½Ð¸Ð¸ Ð´Ð¸Ð¿Ð»Ð¾Ð¼Ð½Ð¾Ð¹ ÑÐ°Ð±Ð¾ÑÑ Ð±ÑÐ»Ð¸ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ñ ÑÐ»ÐµÐ´ÑÑÑÐ¸Ðµ
ÑÐºÑÐ¿ÐµÑÐ¸Ð¼ÐµÐ½ÑÑ: ÐºÑÐ»ÑÑÐ¸Ð²Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ Ð¼Ð°ÐºÑÐ¾ÑÐ°Ð³Ð¾Ð² Ð¿ÐµÑÐ¸ÑÐµÑÐ¸ÑÐµÑÐºÐ¾Ð¹ ÐºÑÐ¾Ð²Ð¸ Ð²
Ð¿ÑÐ¸ÑÑÑÑÑÐ²Ð¸Ð¸ Ð¸ Ð±ÐµÐ· Ð¤Ð¨ Ð°Ð¼Ð±ÑÐ¾ÐºÑÐ¾Ð»Ð°, Ð¾ÑÐµÐ½ÐºÐ° ÑÐµÐ½Ð¾ÑÐ¸Ð¿Ð¸ÑÐµÑÐºÐ¾Ð³Ð¾ ÑÐ¾Ð·ÑÐµÐ²Ð°Ð½Ð¸Ñ
Ð¼Ð°ÐºÑÐ¾ÑÐ°Ð³Ð¾Ð², Ð¾ÑÐµÐ½ÐºÐ° Ð²ÑÐ¶Ð¸Ð²Ð°ÐµÐ¼Ð¾ÑÑÐ¸ Ð¼Ð°ÐºÑÐ¾ÑÐ°Ð³Ð¾Ð² Ð¿ÑÐ¸ Ð´Ð¾Ð±Ð°Ð²Ð»ÐµÐ½Ð¸Ð¸ ÑÐ°Ð·Ð»Ð¸ÑÐ½Ð¾Ð¹
ÐºÐ¾Ð½ÑÐµÐ½ÑÑÐ°ÑÐ¸Ð¸ Ð¤Ð¨ Ð°Ð¼Ð±ÑÐ¾ÐºÑÐ¾Ð»Ð°, Ð¾ÑÐµÐ½ÐºÐ° Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ GCase Ð¸ ÐºÐ¾Ð½ÑÐµÐ½ÑÑÐ°ÑÐ¸Ð¸
HexSph Ð² ÑÑÑÐ¸Ñ Ð¿ÑÑÐ½Ð°Ñ Ð¼Ð°ÐºÑÐ¾ÑÐ°Ð³Ð¾Ð², ÑÑÐ°ÑÐ¸ÑÑÐ¸ÑÐµÑÐºÐ°Ñ Ð¾Ð±ÑÐ°Ð±Ð¾ÑÐºÐ° Ð´Ð°Ð½Ð½ÑÑ
description: The gradient work "Ambroxol influence on the enzymatic glucocerebrosidase
activity in primary macrophage culture" is devoted to evaluating the effect of
pharmacological chaperone (FSH) Ambroxol on changes in the activity of the
glucocerebrosidase enzyme (GCase) and the level of lysosphingolipid
hexosylsphingolysis (HexSph) in dry macrophage spots in the peripheral blood of
patients with Parkinson's disease associated with mutations in the GBA gene
(GBA-BP). In the course of the work, peripheral blood macrophages were cultured
in patients with GBA-BP, as well as in neurologically healthy individuals (control)
in the presence and without Ambroxol, followed by the formation a Bank of dry
macrophage spots in all individuals included in the study for further evaluation of
GCase activity and HexSph concentration by HPLC MS/MS.
The graduate work was performed on the basis of the laboratory of human
molecular genetics of the Kurchatov Institute - PNPI. In the course of this work, it
was shown that human peripheral blood macrophages are a good model for
evaluating changes in GCase activity and the concentration of its substrate when
exposed to drugs for drug testing. For the first time, Ambroxol was shown to
effectively restore GCase activity and reduce HexSph concentration in peripheral
blood macrophages of patients with GBA-BP, which shows the potential use of
this FSH in GBA-BP therapy.
The following experiments were performed during the graduate work: cultivation
of peripheral blood macrophages in the presence and without FSH Ambroxol,
phenotypic characterization of monocytes and macrophages, quantify macrophages
survival, assessment of GCase activity and HexSph concentration in dry
macrophage spots, statistical analysis