ABSTRACT
Autoimmune hepatitis (AIH) is a severe globally distributed liver disease that could occur at any age. Human menstrual blood-derived stem cells (MenSCs) have shown therapeutic effect in acute lung injury and liver failure. However, their role in the curative effect of AIH remains unclear. Here, a classic AIH mouse model was constructed through intravenous injection with concanavalin A (Con A). MenSCs were intravenously injected while Con A injection in the treatment groups. The results showed that the mortality by Con A injection was significantly decreased by MenSCs treatment and liver function tests and histological analysis were also ameliorated. The results of phosphoproteomic analysis and RNA-seq revealed that MenSCs improved AIH, mainly by apoptosis and c-Jun N-terminal kinase/mitogen-activated protein signaling pathways. Apoptosis analysis demonstrated that the protein expression of cleaved caspase 3 was increased by Con A injection and reduced by MenSCs transplantation, consistent with the TUNEL staining results. An AML12 co-culture system and JNK inhibitor (SP600125) were used to verify the JNK/MAPK and apoptosis signaling pathways. These findings suggested that MenSCs could be a promising strategy for AIH.
Subject(s)
Mice , Animals , Humans , Hepatitis, Autoimmune/pathology , Signal Transduction , Disease Models, Animal , Stem CellsABSTRACT
Objective To explore the dosage and injection method of concanavalin A(Con A) for inducing Wistar rats into the acute hepatic injury model. Methods (1)According to the dosage of Con A, 42 Wistar rats were randomly divided into groups A, B, C, D, E, N, 7 rats in each group. Group N was given tail intravenous injection of normal saline as normal control group. Groups A, B, C, D, E were given intravenous injection of 4, 8, 16, 30, 40 mg/kg of Con A respectively. At the 8th hour after modeling, the levels of alanine transaminase(ALT), aspartate aminotransferase(AST), albumin(ALB), interleukin(IL)-2 , IL-10, interferon (IFN)-γ, and tumor necrosis factor(TNF)-αwere detected. And HE staining was used to observe the pathological feature of hepatic tissue. (2)According to the injection method of Con A, 21 Wistar rats were randomly divided into normal control group, intraperitoneal injection group and tail intravenous injection group, 7 rats in each group. The dosage of Con A for the rats in intraperitoneal injection group and tail intravenous injection group was 16 mg/kg. At the 8th hour after modeling, the levels of serum ALT, AST, and ALB were determined. Results The number of abnormal deaths in various dose Con A groups at the end of each experiment was 0 in groups A, B, C, and 2 in group D, and 7 in group E. A small amount of spotty necrosis, inflammatory cell infiltration, and hepatic lobule with almost integrity of structure were found in groups A, B, while obvious bridging-like necrosis was seen in groups C, D. Serum ALT, AST, and ALB levels in intraperitoneal injection group had no statistically significant difference as compared with the normal control group. Conclusion Tail intravenous injection of 16 mg/kg of Con A can be used to induce an acute immunological liver injury rat model successfully.
ABSTRACT
Objective To study the protective effects of aqueous extract and alcohol-soluble extract of Isodon lophanthoides var.gerardianus (Benth.)Hara (ILVG) on immunity hepatic injury induced by concanavalin A (Con A) in mice.Methods One hundred and eight NIH mice were allocated into normal control group,model group,ILVG groups of aqueous extract and alcohol-soluble extract (low-,middle-and high-dosage),and bifendate group randomly.In the experimental groups,mice received either ILVG aqueous extract or alcohol-soluble extract (18.20 g?kg-1,9.10 g?kg-1,4.55 g?kg-1 respectively) or Bifendate (45 mg?kg-1) by gastric perfusion daily for consecutive 5 days.In the 5th day,Con A (20 mg?kg-1) was injected into mice via the tail vein 4h after administration.And then the blood was obtained by picking out the eyeball and the serum was separated after 8-hour fasting.The serum levels of ALT and AST were analyzed,the body weight as well as the weight of liver,spleen and thymus were measured,and pathological features of hepatic tissue were observed.Results ILVG can decrease the ALT and AST,restrain the enlargement of liver and the shrinkage of thymus and reduce the necrosis of hepatic tissue.Conclusion Aqueous extract and alcohol-soluble extract of ILVG possess the effects of protecting liver from immunity injury induced by Con A in NIH mice.