| Objective: Diabetic Nephropathy(DN)is a very serious chronic illness among all other complications of Diabetes Mellitus(DM).DN is the main cause of chronic renal failure in diabetes,and has become one of the leading causes of end-stage renal failure.The pathogenesis of DN was not yet fully elucidated,but its occurrence and development was the result of multi-factor comprehensive effect of high glucose state.Evidence shows that the combination of immunology and inflammatory mechanisms played a pivotal role in its expression,development and eventual progress.Sustained hyperglycemia in diabetic patients could lead to increased glycosylation of end products(AGEs).The binding of AGEs and AGEs receptors stimulates macrophages to synthesize monocyte chemoattractant protein-1(MCP-1)and tumor necrosis Factor(TNF-α)etc.This leads to kidney damage,mesangial cell proliferation,and extracellular matrix proliferation.The expression of MCP-1 in diabetic nephropathy was positively correlated with the infiltration of renal macrophages.Researches show that MCP-1 can promote the migration and activation of mononuclear cells from the peripheral blood to the renal tissue,which leads to the aggregation and promotion of macrophages in the kidney growth factors and inflammatory mediators increase.It also promotes extracellular matrix proliferation,and ultimately causes glomerular sclerosis.There is still a lack of the ideal method and drugs to the prevention and treatment of DN.Astragalus membranaceus has long been known as an immunomodulator in traditional Chinese medicine.In clinical practice,astragalus has been widely used in the treatment of diabetes and the treatment of kidney abnormalities caused by diabetes.In clinical practice,Astragalus is one of the traditional Chinese medicine that can produce a significant reduction in the role of urinary protein.It is commonly used in the prevention and treatment of nephrotic syndrome.However,its regulation of the inflammatory response mechanism is unclear.The current study uses astragalus in db/db mouse model to explore the astragalus immunore-gulatory effect on db/db mice kidney,thus to provide experimental basis for the clinical treatment of DN.Methods:1 Allocation of animals : 10 C57 mice were selected as control group(C).20 db / db mice,4 weeks old,male and female were randomly allocated into the pathogenic group(DM)and the treatment group(DM + H.After one week of environment adaption,the mice from group DM + H were injected with Astragalus depending on their weights(e.g.each mouse was injectioned on intraperitoneal for 0.15 ml astragalus).Mice from control group C and group DM mice were injected with intraperitoneal at 0.15 mlPBS.Astragalus Injection continued for 12 weeks until the mice were sacrificed.Blood samples were collected along with serum.Kidney and spleen tissue were reserved in frozen tube in fridge,with liquid nitrogen fixed at-80℃.2 Detection of the immunological cytology of mice with blood analyzer.3 Detection of blood glucose levels in mice serum.4 Real-time quantitative detection of mRNA expression of IL-1β,IL-10,TNF-α,IL-12β,IFN-β and MCP-1 in mouse kidney,IL-10,IL-4,IL-6 and IFN-γ in mouse spleen.Results:1 The blood glucose levels: Compared with the blank control group(6.7±0.76mmol/L),the blood glucose level of the pathogenic group(23.2±4.73 mmol/L)and the treatment group(24.5±5.02 mmol/L)were both increased significantly,but there was no significant difference observed in the blood glucose level between the treatment group and the pathogenic group.2 Immune cells test results: Compared with the blank control group(4.99±1.21)%,the percentage of mononuclear cells in the pathogenic group(11.02±2.07)% increased significantly,while the percentage of mononuclear cells in the treatment group(4.56±1.02)% was lower than that in the pathogenic group,but no strong enough to produce statistically significant difference.3 mRNA expression of inflammatory factors in mouse kidneyThe mRNA expression of IL-10,IL-1β,MCP-1 and TNF-α in the pathogenic group was significantly higher than that in the control group,the expression in the treatment group was significantly lower than that in the pathogenic group.IL-12β:There was no significant change in the three groups.IFN-β:Compared with the control group,the expression of the pathogenic group showed an increase trend,but there was no statistically significant difference,while the treatment group was lower than the pathogenic group.But statistically significant difference was not observed from any of the three comparisons for mRNA expression.4 mRNA expression of inflammatory factors in spleen of miceIL-4 and IFN-γ:Compared with the control group,the pathogenic group and the treatment group showed a decreasing trend without statistically significant difference.The pathogenic group and the treatment group did not change significantly.IL-6:There was no significant change in the three groups.IL-10:Compared with the control group,the pathogenic group and the treatment group were increased to a certain extent,but no statistically significant difference observed.There was no significant change between the treatment group and the pathogenic group.Conclusion:1 The effect of astragalus on db / db mice blood glucose is not obvious.2 Astragalus had a significant inhibitory effect on IL-10,IL-1β,MCP-1 and TNF-α in db / db mice kidney,but limited effect of IL-4,IFN-γIL-6 and IL-10 on spleen of mice.The results suggest that astragalus have significant anti-inflammatory effect in the kidneys of db/db mice,but the anti-inflammatory effect in spleen of db/db mice is not obvious.3 Astragalus can regulate the number of white cells and monocytes in db / db mice. |
