Đánh giá sự biểu hiện của các gen tự thực (autophagy) trong quá trình hoạt hóa in vitro của các tế bào hình sao từ gan chuột

0 Người đánh giá. Xếp hạng trung bình 0

Tác giả: Xuan Le Holterman Ai, Lê Trâm Anh Cao, Minh Thành Đặng, Thị Tùng Loan Đặng, Quang Huy Đỗ, Minh Huy Lê, Văn Trình Lê, Văn Thuận Nguyễn, Văn Phúc Phạm, Lữ Chính Nhân Phan, Thị Hằng Trần, Hải Nhung Trương

Ngôn ngữ: vie

Ký hiệu phân loại: 574.87 [Unassigned]

Thông tin xuất bản: Khoa học & Công nghệ Việt Nam, 2021

Mô tả vật lý: 21-26

Bộ sưu tập: Metadata

ID: 494398

Thêm vào giỏ

Nghiên cứu này nhằm đánh giá sự thay đổi biểu hiện các gen liên quan đến sự tự thực trong quá trình hoạt hóa của các tế bào hình sao in vitro. Tế bào hình sao phân lập từ gan chuột được nuôi cấy in vitro trong 7 ngày. Mức độ hoạt hóa của tế bào hình sao được đánh giá thông qua hình thái, mức độ dự trữ các giọt mỡ nội bào bằng nhuộm dung dịch oil red o (ORO)
biểu hiện của các gen hoạt hóa α-sma, collagen I và lrat bằng qRT PCR và nhuộm ICC (immunocytochemistry) (α-SMA). Sự biểu hiện của các gen tự thực lc3b, beclin 1, atg12 được đánh giá bằng qRT PCR và nhuộm ICC (LC3). Kết quả cho thấy, tế bào hình sao hoạt hóa khi nuôi cấy in vitro sau 3 và 7 ngày thể hiện thông qua sự thay đổi hình dạng giống nguyên bào sợi cơ, mất dần khả năng tích trữ các giọt mỡ, tăng sự biểu hiện của các gen hoạt hóa α-sma và collagen I, giảm sự biểu hiện của gen lrat. Thêm vào đó, sự biểu hiện của các gen tự thực lc3b, beclin 1 và atg12 tăng lên ở các tế bào hình sao hoạt hóa khi nuôi cấy in vitro sau 3 và 7 ngày. Đây là kết quả ban đầu trong nghiên cứu tìm ra vai trò của sự tự thực trong quá trình hoạt hóa của tế bào hình sao, hướng tới các liệu pháp điều trị xơ gan nhắm vào sự tự thực.This study aims to evaluate the expression level of autophagy genes during the activation of mouse hepatic stellate cells (HSCs) in vitro. The HSCs isolated from the mouse liver were cultured in vitro for 7 days. The activation of HSCs was evaluated by their morphology, the storage of lipid droplets by oil red O (ORO) staining, the expression of activation-related genes α-sma, collagen I, and quiescence-related gene lrat by qRT PCR and ICC staining (α-SMA). The expression of autophagy genes lc3b, beclin 1, atg12 were assessed by qRT PCR and ICC staining (LC3). The results showed that the isolated HSCs were activated after 3 days and 7 days of the culture in vitro. The activation was indicated by the morphological change of HSCs to myofibroblast-like cells, loss of lipid droplets, and increased expression of fibrotic genes α-sma and collagen I, decreased expression of lrat. Additionally, the expression of autophagy genes lc3b, beclin 1, and atg12 were significantly increased in the activated HSCs after the culture in vitro for 3 and 7 days. This study contributes the preliminary results to further studies on the role of autophagy during the activation of HSCs, which may be exploited for the development of the antifibrotic therapy targeting autophagy.

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