Qin Q, Li R, Zhou Y, Zhang Y, Han M, Zhu L
Peking University Ditan Teaching Hospital, Beijing 100015, China.
Beijing Key Laboratory of Emerging Infectious Diseases, Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2025 Aug 18;57(4):727-734. doi: 10.19723/j.issn.1671-167X.2025.04.016.
To explore the protective effect of knock-down the expression of B lymphocyte induced maturation protein 1 () gene on early liver injury in carbon tetrachloride (CCl)-induced mouse model of liver fibrosis.
C57BL/6 mice were intraderitoneal injected with 5% CCl olive oil solution to create mouse model of hepatic fibrosis. The expression of 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 (=10), CCl+AAV-shRNA-NC group (=10) and CCl+AAV-shRNA-Blimp1 group (=10). After 27 days of preparation of the CCl 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.
The expression levels of Blimp1 protein in the liver of CCl+AAV-shRNA-NC group (2.036±0.244, =3.690, =0.002) were significantly increased than that of the blank test group. In the CCl+AAV-shRNA-Blimp1 group, the expression of Blimp1 protein decreased to the basal level (0.783±0.249, =6.223, =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 CCl+AAV-shRNA-NC group, the serum levels of ALT [(894.0±360.1) U/L, =3.998, =0.003] and AST [(820.0±100.6) U/L, =6.141, =0.004] in the CCl+AAV-shRNA-Blimp1 group were significantly decreased. The pathological results of the CCl+AAV-shRNA-Blimp1 group showed that compared with the CCl+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, =7.930, =0.001), COL1A1 (1.426±0.143, =6.364, =0.003) and COL3A1 (1.124±0.198, =3.440, =0.026) of liver in the CCl+AAV-shRNA-Blimp1 group were significantly decreased than that of CCl+AAV-shRNA-NC group, and the mRNA expression levels were altered as well as their protein levels.
Blimp1 plays an important role in CCl-induced liver fibrosis in mice, and knock-down the expression of gene is beneficial to protect early liver injury in mice.
探讨敲低B淋巴细胞诱导成熟蛋白1（Blimp1）基因表达对四氯化碳（CCl₄）诱导的小鼠肝纤维化早期肝损伤的保护作用。
将C57BL/6小鼠腹腔注射5% CCl₄橄榄油溶液以建立肝纤维化小鼠模型。通过腹腔注射短发夹RNA（shRNA）腺相关病毒（AAV）降低小鼠体内Blimp1基因的表达。将小鼠随机分为3组：空白对照组（n = 10）、CCl₄ + AAV-shRNA-NC组（n = 10）和CCl₄ + AAV-shRNA-Blimp1组（n = 10）。在制备CCl₄小鼠模型27天后，进行取材。采用蛋白质免疫印迹法（Western blot）和实时荧光定量聚合酶链反应（real-time PCR）检测各组肝组织中Blimp1、α-平滑肌肌动蛋白（α-SMA）、Ⅰ型胶原蛋白α1（COL1A1）、Ⅲ型胶原蛋白α1（COL3A1）的水平及其mRNA表达水平。分离各组小鼠血清，用全自动生化分析仪检测天冬氨酸转氨酶（AST）和丙氨酸转氨酶（ALT）。通过苏木精-伊红染色、Masson染色和天狼星红染色等病理染色检测小鼠肝组织的病理变化及肝纤维化程度。
CCl₄ + AAV-shRNA-NC组小鼠肝脏中Blimp1蛋白表达水平（2.036±0.244，t = 3.690，P = 0.002）明显高于空白对照组。在CCl₄ + AAV-shRNA-Blimp1组中，Blimp1蛋白表达降至基础水平（0.783±0.249，t = 6.223，P = 0.003）。与CCl₄ + AAV-shRNA-NC组血清ALT水平[（1 957.8±633.6）U/L]和AST水平[（1 808.8±260.1）U/L]相比，CCl₄ + AAV-shRNA-Blimp1组血清ALT水平[（894.0±360.1）U/L，t = 3.998，P = 0.003]和AST水平[（820.0±100.6）U/L，t = 6.141，P = 0.004]明显降低。CCl₄ + AAV-shRNA-Blimp1组病理结果显示，与CCl₄ + AAV-shRNA-NC组相比，肝组织中炎症细胞浸润减少，纤维化程度减轻。CCl₄ + AAV-shRNA-Blimp1组肝脏中α-SMA（0.676±0.064，t = 7.930，P = 0.001）、COL1A1（1.426±0.143，t = 6.364，P = 0.003）和COL3A1（1.124±0.198，t = 3.440，P = 0.026）水平明显低于CCl₄ + AAV-shRNA-NC组，其mRNA表达水平和蛋白水平均发生改变。
Blimp1在CCl₄诱导的小鼠肝纤维化中起重要作用，敲低Blimp1基因表达有利于保护小鼠早期肝损伤。
