Effects of Saikosaponin d on differentially expressed genes CTLA-4, IL-10 and IL-17 in mice with autoimmune hepatitis
2020, 40 (3):
Objective · To explore the effects of Saikosaponin d (SS-d) on autoimmune hepatitis (AIH)observing the 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 (n8) and SS-d treatment group (n32). 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 n4). The mice were sacrificed after 12 h. The differentially expressed genes of liver were screened, some of which were verifiedqRT-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 n8). 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 detectedELISA, and cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) was detectedWestern 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 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 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 miceregulating the of IL-10, CTLA-4, and IL-17.
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