敲低多房棘球蚴let-7-5p可抑制BALB/c小鼠腹腔巨噬细胞极化和虫体生长

doi:10.11843/j.issn.0366-6964.2024.06.033

Abstract

Abstract:

The purpose of this study was to determine the effects of knockdown of Echinococcus multilocularis emu-let-7-5p on mouse peritoneal macrophages polarization and worm growth. A recombinant adeno-associated virus vector expressing emu-let-7-5p sponge sequence of E. multilocularis was constructed, and its inhibitory effect and specificity were validated in 293T cells. Seventy clean-grade BALB/c mice were injected intravenously (tail vein) with AAV8-si-let-7-5p, AAV8-control, and PBS, respectively. After two weeks, two mice in each group were randomly selected, and the expression of GFP, emu-let-7-5p and its target C/EBP δ in liver were detected by Western blot and qRT-PCR. At 3 months after intraperitoneal inoculation of 1 000 protoscolexs in each mouse, the transcriptional levels of emu-let-7-5p, C/EBP δ, M1 type (IL-1β, iNOS and IL-6) and M2 type (Arg-1, IL-4 and IL-10) related molecules in peritoneal macrophages were detected by qRT-PCR, and cysts weight and protoscolexs number of E. multilocularis in each group were analyzed. The results showed that, on the 15th day after injection, AAV8-si-let-7-5p successfully targeted the mouse liver and caused knockdown of emu-let-7-5p and up-regulation of C/EBP δ. At 3 months after infection, the expressions of emu-let-7-5p and M2 related molecule (Arg-1) in peritoneal macrophages were significantly down-regulated in AAV8-si-let-7-5p group, while C/EBP δ and M1 related molecules (IL-1β and iNOS) were significantly up-regulated. In addition, in AAV8-si-let-7-5p group, the expressions of IL-4 and IL-6 were down-regulated, while the expression of IL-10 was up-regulated, and cysts weight and protoscolexs number in AAV8-si-let-7-5p group were significantly lower than those in the control group. The above results suggested that knockdown of emu-let-7-5p in mice infected with E. multilocularis can promote the expression of C/EPB δ, and thus inhibit M2 macrophage polarization and worm growth.

Key words: Echinococcus multilocularis, emu-let-7-5p, peritoneal macrophages polarization, worm growth

CLC Number:

S852.734

Liqun WANG, Yixuan WU, Guiting PU, Shanling CAO, Dexian WNAG, Tingli LIU, Hong LI, Tharheer Oluwashola AMUDA, Xiaola GUO, Hong YIN, Xuenong LUO. Knockdown of let-7-5p from Echinococcus multilocularis Inhibited the Peritoneal Macrophages Polarization and Worm Growth in BALB/c Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(6): 2619-2628.

Figures/Tables 7

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References 30

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引物名称 Primer name	上游引物(5′→3′) Forward primer	下游引物(5′→3′) Reverse primer
U6	CTCGCTTCGGCAGCACA	AACGCTTCACGAATTTGCGT
let-7-5p	AGCTGGACTGAGGTAGTGTTTCG	ATCCAGTGCAGGGTCCGAGG
GAPDH	AGGTCGGTGTGAACGGATTTG	TGTAGACCATGTAGTTGAGGTCA
C/EBP δ	CGACTTCAGCGCCTACATTGA	CTAGCGACAGACCCCACAC
IL-1β	AGAGCATCCAGCGTCAAATC	TCATCTCGGAGCCTGTAGTG
IL-6	CTTCTTGGGACTGATGCTGGTGAC	TCTGTTGGGAGTGGTATCCTCTGTG
iNOS	CGAAACGCTTCACTTCCAA	TGAGCCTATATTGCTGTGGCT
IL-4	GCTATTGATGGGTCTCACCC	CAGGACGTCAAGGTACAGGA
IL-10	GCTCTTACTGACTGGCATGAG	CGCAGCTCTAGGAGCATGTG
Arg-1	TCATGGAAGTGAACCCAACTCTTG	TCAGTCCCTGGCTTATGGTTACC

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[2]	LI Hong, LI Yanping, LIU Tingli, CHEN Guoliang, WANG Liqun, GUO Xiaola, LUO Xuenong. The Effect of Echinococcus multilocularis Infection on Lipid Metabolism in the Liver of Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 263-271.
[3]	LI Yanping, LIU Tingli, LI Hong, CHEN Guoliang, WANG Liqun, GUO Xiaola, LUO Xuenong. Construction and Analysis of Polarization-related ceRNA Regulatory Network in Mouse Kupffer Cells Induced by Echinococcus multilocularis [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4410-4418.

Knockdown of let-7-5p from Echinococcus multilocularis Inhibited the Peritoneal Macrophages Polarization and Worm Growth in BALB/c Mice

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