Effects Of Artificial Cordyceps Sinensis On The Liver And Mechanisms In STZ Induced Diabetic Rats

Object:Diabetic patients exist multiple organ lymphatic cells invasion and immune complex deposition,prompt the formation of diabetic chronic complications have a variety of abnormal immune mechanisms involved, speculated that diabetic liver disease also may exist abnormal immune and immune damage. Diabetic liver disease(DLD) is one of the most important diabetic chronic complications of diabetes mellitus(D). In addition, a low degree of inflammation is increasingly Nsidered to the major mechanisms which Ntribute to the development of liver damage in diabetes. Elevated glutamic-pyruvic transaminase is the most important presentation of DLD. Hepatocellular fatty degeneration,necrosis,hepatic Kuffer cells and hepatic stellate cell activation, abnormal secretion of extracellular matrix,the cause of liver fibrosis. Monocyte chemoattractant protein-1(MCP-1) is a potent chemokine, which can induce the aggregation of various inflammatory cells, especially monocytes. The existed researches showed that the MCP-1texcessive chemotaxis of mononuclear cells that cause intrahepatic inflammation may plays a key role in the process of diabetic liver disease.TNF-a is a single-core factor,mainly produced by macrophages and monocytes. TNF-α also can produced by neutrophils, Kuffer cells,endothelial cells,smooth muscle cells. TNF-a is mainly produced by Kuffer cells in the liver. TNF-a have complicated biological activities,including antitumor,antivirus,immunomodulatory and induce inflammatory reaction and so on.It played important roles in liver fibrosis development.IL-6is secreted by mononuclear macrophages and T lymphocytes,it is the cell factors that have some nonspecific immunity function and play an important role in the inflammatory response,its role has aroused many scholars attention which play a important role in the physiological and pathological process of liver.The IL-6of liver is mainly secreted by activated hepatic stellate cells and hepatic.The degree of liver necrosis and liver fibrosis is closely related with IL-6and TNF-a.IL-6and TNF-a participate in the liver damage-repair cycle in the development of the hepatic cirrhosis,eventually cause liver synthesis and deposition of extracellular matrix,which can lead to liver fibrosis.Pglitazone, one of the thiazolidinediones(TZDs), which plays a role in the liver protective effect through a variety of ways, including regulating glucose and lipid metabolism, improving insulin resistance, improving inflammation and atherosclerosis. Cordyceps sinensis (Berk.) Sacc is a kind of valuable, traditional and invigorating the kidney herb in our country. It is Nfirmed that Cordyceps sinensis (Berk.)Sacc has the effect of reinforcing on kidneys and lungs, improving fragile Nditions, and assisting pneuma. Previous studies found that CS has some therapeutic effects on the liver fibrosis, but the exact mechanism is not clear.In this study, Type1diabetic rat model was established by intravenous injection of streptozocin (STZ). Cordyceps sinensis cultivated by artificial fermentation and Pglitazone were applied to the diabetic rats. The expression of MCP-1、TNF-α and IL-6of the hepatic were detected by the use of pathological morphology and molecular biology. Our aim was to assess the potential relevance of the MCP-1TNF-a and IL-6in the occurrence and development of diabetic liver disease. At the same time, the hepatic protective mechanism of Cordyceps sinensis cultivated by artificial fermentation and Pglitazone were investigated in this study.Materials and Methods:90eight-week male SD rats were divided into two groups randomly:10normal Ntrol rats (group N) and80experimental rats. The D rat models were made by intravenous injection of STZ(45mg/kg).Then, they were randomly assigned into six groups, including Ntrol group(group N),diabetes group(group D), small-dose CS group(group L,0.6g/kg/d), medium-dose CS group(group M,2.5g/kg/d),large-dose CS group(group H,5g/kg/d), P treatment group(group P,4mg/kg/d). Eight weeks later, liver function, total cholesterol and triglyceride were evaluated. The Pathological changes of the liver were studied by Light Microscope. The expression of MCP-1、TNF-α and IL-6were detected by immunohistochemistry. The mRNA expressions of MCP-1、TNF-α and IL-6in liver tissue was assessed by reverse transcription-polymerase chain reaction(RT-PCR).Results:1.In group D,multifocal inflammatory cells invasion,fatty degeneration of liver cells visible,partial hepatic sinus expansion visible. The changes mentioned above were relieved to some extent in the groups treated with P or CS. Furthermore, the effects in group H and P were better than those in groups L or M (P＜0.05).2. In group N,in Massin dyeing, light blue-green collagen only can be stained around the blood vessels and bile ducts, but it can not be stained in the liver parenchyma. But in group D,large amounts of collagen deposition can be seen in the liver parenchyma,and present diffuse distribution,the collagen deposition of vessels is incrassation. The changes mentioned above were relieved to some extent in the groups treated with P or CS,the degree of liver fibrosis is significantly improved. Furthermore, the effects in group H and P were better than those in groups L or M (P＜0.05).3. Comparing with the Ntrol group, the expression of MCP-1was remarkably increased in group D.The expression of MCP-1was obviously decreased in P or CS treatment groups than in group D. MCP-1were significantly lower in group H and P than those in groups L or M (P＜0.05).4. Comparing with the Ntrol group, the expression of TNF-a was remarkably increased in group D.The expression of TNF-a was obviously decreased in P or CS treatment groups than in group D. TNF-a were significantly lower in group L and P than those in groups H or M (P＜0.05).5. Comparing with the Ntrol group, the expression of IL-6was remarkably increased in group D.The expression of IL-6was obviously decreased in P or CS treatment groups than in group D. IL-6 were significantly lower in group H and P than those in groups L or M (P＜0.05).Nclusions:1.Our data has shown abnormal liver function, hepatocyte fatty degeneration,the expression of MCP-1、TNF-α and IL-6was increased obviously in STZ induced diabetic rats.2. In diabetic rats, the expression of MCP-1TNF-a and IL-6was remarkably increased,and there was a closely correlation between MCP-1、 TNF-α、IL-6and liver fibrosis.3. Our data has shown the liver protective effect of both CS and P on diabetic liver disease. the mechanisms may be related to inhibit the expression of MCP-1,TNF-a and IL-6. The renal protective effects seems to be independent of blood glucose Ntrol.