Effects of Saikosaponin d on differentially expressed genes CTLA-4, IL-10 and IL-17 in mice with autoimmune hepatitis
10.3969/j.issn.1674-8115.2020.03.005
- Author:
Jian-Heng HAO
1
Author Information
1. School of Basic Medical Sciences, Shanxi University of Traditional Chinese Medicine
- Publication Type:Journal Article
- Keywords:
Autoimmune hepatitis (AIH);
Cytotoxic T lymphocyte-associated antigen-4 (CTLA-4);
Interleukin-10;
Interleukin-17;
Saikosaponin d (SS-d)
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2020;40(3):303-309
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To explore the effects of Saikosaponin d (SS-d) on autoimmune hepatitis (AIH) by observing the expression changes of some differentially expressed genes screened with the Agilent-085631 gene chip in the liver of AIH mice. Methods: Forty healthy male SPF C57BL/6 mice were divided into chip group (n=8) and SS-d treatment group (n=32). The mice in the chip group were divided into the normal group and the model group [concanavalin A (Con A) was administered to the tail vein at a dose of 15 mg/kg] (both n=4). The mice were sacrificed after 12 h. The differentially expressed genes of liver were screened, some of which were verified by qRT-PCR. The SS-d treatment group was further divided into the normal group, the model group (treatment was the same with the chip group), SS-d low-dose group and SS-d high-dose group [according to the literature and pre-experiment results, 2.5 and 5.0 mg/kg dose of intraperitoneal injection of SS-d were given respectively, and 15 mg/kg of Con A was administered to the tail vein 8 h later] (all n=8). After 12 h, total venous blood, liver total protein and total RNA of mice were collected. The levels of serum glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT), interleukin (IL)-10 and IL-17 were detected by ELISA, and cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) was detected by Western blotting. qRT-PCR technology was used to detect the mRNA levels of IL-10, IL-17 and CTLA-4. Results: A total of 11 512 differentially expressed genes were screened (up 5 189, down 6 323), which were related to 138 signal pathways. The qRT-PCR results of IL-10, IL-17 and CTLA-4 gene were consistent with the results of chip screening. Compared with the normal group, the serum levels of GPT and GOT in the model group increased, IL-17 mRNA level increased, IL-10 mRNA and CTLA-4 mRNA levels decreased, the content of serum IL-17 increased, the content of serum IL-10 decreased, and the level of CTLA-4 protein expression in the liver tissues decreased. Compared with the model group, the serum GPT and GOT levels of SS-d in the low-dose and high-dose groups decreased, IL-17 mRNA level decreased, the levels of IL-10 mRNA and CTLA-4 mRNA increased, the content of serum IL-17 decreased, the content of serum IL-10 increased, and the level of CTLA-4 protein expression in the liver tissue increased. Conclusion: Multiple signaling pathways are involved in the pathogenesis of AIH, and SS-d can alleviate the liver inflammation in AIH mice by regulating the expression of IL-10, CTLA-4, and IL-17.
