Effect Of DMOC Prescription On Liver Damage And Hepatic Fibrosis In Diabetic Model Rats By JAK2/STAT5/PPARγ Pathway

Objective:To study the effects of DMOC on the expressions of the Janus kinase/signal transduction and transcription activation of transcription (JAK/STAT) signaling pathway in streptozotocin induced diabetic rat with hepatic injury or hepatic fibrosis.To assess the partly mechanism of protective effect on the liver function, decreasing liver fibrosis by DMOC.Methods:After a one-week adaptive feeding,60 female Wistar rats (body weight 200g 200 ± 20g,SPF level),were randomly divided into normal group(Control,n=10,fed with normal diet) and model established group(n=50).Type 2 diabetes with hepatic injury or hepatic fibrosis model rats were induced by intraperitoneal injection of Streptozoein(STZ,25 mg/kg+m2,interval 2 days) after high glucose and high fat diet for 4 weeks.After the modeling,rats measured twice in fasting blood glucose>11.lmmol/L were randomly divided into 3 groups:Model group(Model,n=10,fed with normal diet andlg(kg.d) ddH2O),DMOC treatment group[DMOC,n=10,fed with normal diet and lOg (kg.d) DMOC]; metforrnin treatment group[DMBG,n=10.fed with normal diet and 500 mg/(kg/d) metformin];normal control group[Control,n=10,fed with normal diet and 10g(kg.d) ddH2O]. Except normal control group,Con A at a dose of 25 mg/kg was injected into abdominal cavity weekly for 9 weeks.The rat incontrol group were injected with PBS instead of Con A. After treatment for 22 weeks,the expression levels of JAK2, STAT5,RXRa,PPARy mRNA and proteins in liver were measured using Real-time PCR,Immunohistochemistry method and western blot.The liver indexes were measured and pathological changes of the liver were observed.Results:1 The condition of rats of each group:1 rats of the model group were died during the experiment, the rest of the groups were without death.The condition of control group was fine,such as,eating normally, moving agilely. They were having bright fur,steady up weight and granular droppings.The rest of rats show different levels of symptoms, such as eating more, drinking more, dark fur,weight gaining slow and diarrhea.After treatment,The rats’ condition of DMOC group and DMBG group became better.2 The fasting blood glucose of each group:fasting blood glucose in rats after modeling (before drug treatment),compared with the normal group,model group,DMBG and DMOC group, fasting blood glucose levels were significantly increased and was statistically significant (P<0.01),the model group,DMBG group and DMOC group in each of the two groups had no difference between fasting blood glucose (P>0.05);After drug intervention (22 weeks),compared with the model group,DMBG group,fasting blood glucose significantly decreased DMOC group,there was significant difference (P<0.01).3 The liver function of each group:before drug treatment, compared with the normal group,the rest groups’ ALT were significantly increased,the difference is statistically significant (P<0.05),, model group,DMBG group and DMOC group between every two groups,the levels of ALT and AST are no difference (P>0.05).After Drugs prognosis (22 weeks),DMOC group’s ALT and AST decreased significantly (P<0.01).model group compared with normal group,ALT decreased,AST increased,but the difference was not statistically significant (P>0.05;compared with the normal group ratio, DMOC group’s ALT,AST significantly lower (P>0.05) and ALT increased,,but the difference was not statistically significant (P>0.05),the difference of AST is statistically significant (P<0.05); DMOC of sequential group, the levels of ALT and AST were low,but only differences in AST have statistical significance (P<0.01).4 Hyaluronic acid and laminin of each group:after drug intervention (22 weeks),compared with the normal group,model group’s HA and LN increased significantly, with statistical significance (P<0.01);compared with the model group, DMOC group’s HA and LN were decreased,there was statistical significant differences (P<0.05),compared with the normal group,DMOC group and DMBG group’s HA are higher but the difference was not statistically significant (P>0.05),compared with the normal group,the LN of DMOC group, but the difference was not statistically significant (P>0.05),DMBG group LN,the difference was statistically significant ((P<0.01).5 Liver tissue Masson three staining:in normal group,, only a small amount of collagen expression in the central vein and portal area. The liver tissue of the model group in portal area and collagen fiber deposition.Compared with the model group, DMOC group and DMBG group periportal collagen deposition was significantly reduced and DMBG group improved particularly significant.6 PPARγ immunohistochemistry:compared with the model group,normal group PPAR gamma protein positive staining was nuclear uniform dark brown,in liver tissue distribution widely;in the model group, the nucleus almost no positive staining;compared with the model group,DMOC group’s positive cells increased significantly,but compared to the normal cells were positive staining shallower.7 The mRNA relative expression of JAK2,STAT5 and PPAR gamma:After drug intervention (22 weeks),compared with the normal group,in model group,JAK2,STAT5and PPAR gamma expression decreased significantly, statistical significance (P<0.01); compared with the model group,JAK2,STAT5 and PPAR gamma expression of DMOC group increased,the difference is statistically significant (P<0.05).With the normal group, JAK2 and PPAR gamma expression quantity is lower, the difference was statistically significant (P<0.01), the increased expression of STAT5 and statistical significance (P<0.01).8 Western blot results ofJAK2, STAT5, RXRa,and PPAR gamma Results:compared with the normal group,model group,the expression of Collagen ions increased,the difference was statistically significant (P<0.05),the expression of T-JAK2,T-STAT5,P-STAT,PPAR, RXRa. The PPAR gamma decreased statistically significant differences (P<0.05),P-JAK2 expression decreased, but no statistically significant difference (P>0.05;compared with the model group,the expression of DMOC group Collagen ions was significantly decreased,the difference was statistically significant (P<0.01);the expression of T-JAK2 and P-STAT5 in DMOC group,RXRa increased significantly,the difference was statistically significant(P<0.01),but compared with the normal group, the expression was lower, the difference was statistically significant (P<0.05),P-JAK2,PPAR,gamma DMOC group expression increased,the difference was statistically significant (P< 0.05).Conclusion:1 DMOC improves blood glucose levels in diabetic patients with liver injury and liver fibrosis induced by high fat and high glucose ConA combined with+STZ.2 DMOC can improve the high fat and high glucose+STZ combined with ConA in diabetic patients with liver injury and liver fibrosis model rats function index ALT, AST, liver fibrosis index HA, LN, reduce the collagen deposition in rat liver.3 DMOC may up regulate expression of JAK2/STAT5/PPAR gamma,improved liver injury and hepatic fibrosis in diabetic rats.