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Journal of Peking University (Health Sciences) ›› 2025, Vol. 57 ›› Issue (4): 727-734. doi: 10.19723/j.issn.1671-167X.2025.04.016

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Protective effect of knock-down the expression of Blimp1 gene on early liver injury in CCl4-induced mouse model of liver fibrosis

Qiushi QIN1, Rui LI2,3, Yanxi ZHOU2,3, Yue ZHANG2,3, Ming HAN2,3, Liuluan ZHU1,2,3,*()   

  1. 1. Peking University Ditan Teaching Hospital, Beijing 100015, China
    2. Beijing Key Laboratory of Emerging Infectious Diseases, Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China
    3. Beijing Institute of Infectious Diseases, Beijing 100015, China
  • Received:2022-09-16 Online:2025-08-18 Published:2025-08-02
  • Contact: Liuluan ZHU
  • Supported by:
    the National Natural Science Foundation of China(81871586); the National Natural Science Foundation of China(81671940)

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Abstract:

Objective: To explore the protective effect of knock-down the expression of B lymphocyte induced maturation protein 1 (Blimp1) gene on early liver injury in carbon tetrachloride (CCl4)-induced mouse model of liver fibrosis. Methods: C57BL/6 mice were intraderitoneal injected with 5% CCl4 olive oil solution to create mouse model of hepatic fibrosis. The expression of Blimp1 gene in the mice was reduced by intraderitoneal injection of short hairpin RNA (shRNA) adeno-associated virus (AAV). The mice were randomly divided into 3 groups: blank test group (n=10), CCl4+AAV-shRNA-NC group (n=10) and CCl4+AAV-shRNA-Blimp1 group (n=10). After 27 days of preparation of the CCl4 mouse model, animal materials were carried out. Western blot and real-time PCR were used to detect the levels of Blimp1, α-smooth muscle actin (α-SMA), collagen type Ⅰ alpha 1 (COL1A1), collagen type Ⅲ alpha 1 (COL3A1), and their mRNA expression levels of liver tissue in each group. The serum of each group was separated to measure aspartate transaminase (AST) and alanine transaminase (ALT) by automatic biochemical analyzer. The pathological changes of liver tissue and the degree of liver fibrosis in the mice were detected by pathological staining including hematoxylin-eosin staining, Masson, and Sirius red. Results: The expression levels of Blimp1 protein in the liver of CCl4+AAV-shRNA-NC group (2.036±0.244, t=3.690, P=0.002) were significantly increased than that of the blank test group. In the CCl4+AAV-shRNA-Blimp1 group, the expression of Blimp1 protein decreased to the basal level (0.783±0.249, t=6.223, P=0.003). Compared with the serum levels of ALT [(1 957.8±633.6) U/L] and AST [(1 808.8±260.1) U/L] in the CCl4+AAV-shRNA-NC group, the serum levels of ALT [(894.0±360.1) U/L, t=3.998, P=0.003] and AST [(820.0±100.6) U/L, t=6.141, P=0.004] in the CCl4+AAV-shRNA-Blimp1 group were significantly decreased. The pathological results of the CCl4+AAV-shRNA-Blimp1 group showed that compared with the CCl4+AAV-shRNA-NC group, the infiltration of inflammatory cells in the liver tissue was reduced and the degree of fibrosis was alleviated. The level of α-SMA (0.676±0.064, t=7.930, P=0.001), COL1A1 (1.426±0.143, t=6.364, P=0.003) and COL3A1 (1.124±0.198, t=3.440, P=0.026) of liver in the CCl4+AAV-shRNA-Blimp1 group were significantly decreased than that of CCl4+AAV-shRNA-NC group, and the mRNA expression levels were altered as well as their protein levels. Conclusion: Blimp1 plays an important role in CCl4-induced liver fibrosis in mice, and knock-down the expression of Blimp1 gene is beneficial to protect early liver injury in mice.

Key words: Liver fibrosis, Liver injury, Animal models, B lymphocyte induced maturation protein 1

CLC Number: 

  • R575.2

Table 1

The sequences of shRNA-Blimp1 and shRNA-NC"

shRNA Oligonucleotides (5′-3′)
shRNA-NC CCGGCAACAAGATGAAGAGCACCAACTCGAGTTGGTGCTCTTCATCTT
shRNA-Blimp1-1 AAAAGGTGCAGCCTTTATGAGTCCTCGAGGACTCATAAAGGCTGCACC
shRNA-Blimp1-2 AAAACTCTCGACAGCAAATGGTTCTCGAGAACCATTTGCTGTCGAGAG
shRNA-Blimp1-3 AAAAGCAGGATTACCCAAGAATACTCGAGTATTCTTGGGTAATCCTGC

Figure 1

Expression levels of Blimp1 in RAW264.7 cells(A, B) and the liver of mice(C, D) The interference efficiency of three shRNA-Blimp1 in RAW264.7 cells and the expression levels of Blimp1 in the liver of blank test, CCl4+AAV-shRNA-NC and CCl4+AAV-shRNA-Blimp1 group mice (n=10). *P < 0.05, **P < 0.01, ***P < 0.001. shRNA, short hairpin RNA; Blimp1, B lymphocyte induced maturation protein 1; CCl4, carbon tetrachloride; AAV, adeno-associated virus; NC, non-specific control; CCl4-sh-NC, CCl4+AAV-shRNA-NC group; CCl4-sh-Blimp1, CCl4+AAV-shRNA-Blimp1 group."

Figure 2

The dynamic changes of body weight in each group mice during modeling The dynamic changes of body weight in the blank test, CCl4+AAV-shRNA-NC and CCl4+AAV-shRNA-Blimp1 group mice (n=10). Abbreviations as in Figure 1."

Figure 3

The ALT and AST levels of serum in each group mice The ALT(A) and AST(B) levels of serum in the blank test, CCl4+AAV-shRNA-NC and CCl4+AAV-shRNA-Blimp1 group mice (n=10). **P < 0.01, ***P < 0.001, ****P < 0.000 1. AST, aspartate transaminase; ALT, alanine transaminase; Other abbreviations as in Figure 1."

Figure 4

Pathological staining of liver tissue in each group Pathological staining of liver tissue in the blank test, CCl4+AAV-shRNA-NC and CCl4+AAV-shRNA-Blimp1 group mice (n=10). The yellow arrows represent inflammatory infiltration, the black arrows represent collagen deposition, and the blue arrows represent liver fibrosis. HE, hematoxylin-eosin staining; Other abbreviations as in Figure 1."

Figure 5

The α-SMA, COL1A1 and COL3A1 levels in liver of mice in each group A and B, the α-SMA, COL1A1 and COL3A1 levels in liver of mice in each group (n=10); C, the mRNA levels of Acta2, Col1a1 and Col3a1 in liver of mice in each group (n=10). * P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.000 1. The Acta2, Col1a1 and Col3a1 is the coding gene of α-SMA, COL1A1 and COL3A1, respectively. Abbreviations as in Figure 1."

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