The Effect Of Chinese Herbal Compound On The Expression Of TGF-?1 And PTEN Protein In The Kidney Tissue Of DN Mice And The Observation Of Its Clinical Efficacy

Purpose:The literature review of this paper was about the clinical manifestations,epidemiology,risk factors,pathology,pathogenesis and treatment of diabetic nephropathy.On the basis of literature review,the combination of clinical trials and basic experimental research were the the methods used to conduct this study.The aim of basic experimental research was to explore the effects of modified prescription of Jian Ling Decoction and modified prescription of Jin Gui Shen Qi wan on blood glucose,urinary protein,renal hypertrophy index,inflammation factors in STZ-induced diabetic models in mice.Besides,histopathological analysis including periodic acid-Schiff(PAS)and immunohistochemical staining for urokinase-type plasminogen activator(u PA),transforming growth factor-?1(TGF-?1)and PTEN,which can down regulate PI3K-PKB / Akt pathway.The traditional Chinese medicine composed of modified prescription of Jian Ling Decoction and modified prescription of Jin Gui Shen Qi wan for the improvement and molecular mechanisms of diabetic nephropathy was clarified in the study.Clinical trials were done to determinated the efficacy and safety of modified prescription of Jin Gui Shen Qi wan as the treatment of diabetic nephropathy.Material and method:1.experimental models of STZ-induced diabetic nephropathy in miceThe 45 C57 BL,male,5 weeks old,mice were fed by high fat diets for 3 weeks.8 weeks old mice were used and randomly divided into 5 groups with group 1 as normal control group consisted of 9 mice.Other groups were designed to produce mouse models of diabetes by intravenously injection of STZ.Each group consisted of 9 mice after successful modeling.Group 2 was DC group,as diabetic control group,did not reviewed any treatment.CM1 group,as traditional Chinese medicine experimental group 1,were given modified prescription of Jian Ling Decoction.CM2 group,as traditional Chinese medicine experimental group 2,were given modified prescription of Jin Gui Shen Qi wan.ACEI group,as Western medicine control group,were given captopril.The body weight and blood glucose of the mice were measured at week 0,the second week and the fourth week after modeled.In addition,experiment was designed to evaluate the effect of TCM on urinary total protein,creatinine and other biochemical indexes of STZ-induced diabetic mice at fourth week after modeled.Death penalty was performed by using carbon dioxide at the 4th week after modeling,and the renal hypertrophy index was calculated by removing kidneys.The effects of the tested medicine on the levels of transforming growth factor-?1(TGF-?1),a protein related with extracellular matrix synthesis,the changes of the inflammatory markers IL-6,as well as urokinase-type plasminogen activator,associated with the thrombolytic system,and PTEN which can negatively regulate the Akt / PKB signaling pathway were observed by immunohistochemistry staining.Therefore,the influences on histopathological change and the above proteins of kidney by comparing with,model group,blank control group,two traditional Chinese medicine experimental groups and positive control group of Western medicine could be determined.2.clinical trialsThe patients enrolled in this study came from the Yusheng Chinese medicine clinic or the division of nephrology at co-operative hospital either inpatient or outpatient from September 2014 to September 2016.All enrolled patients had diabetic nephropathy in stage 3 or 4 by the definition of Mogensen stages of diabetic nephropathy and were characterized with syndrome of Spleen Yang deficiency,Kidney Yang deficiency and Phlegm-damp stagnation.In addition to the original treatment,patients needed to take the modified prescription of Jin Gui Shen Qi wan for 12 weeks.Measurements of clinical symptoms,blood glucose,glycosylated hemoglobin,serum creatinine,urinary creatinine,urinary microalbumin,liver function(AST,ALT)were made before and after the treatment.Results:1.experimental models of STZ-induced diabetic nephropathy in miceThe effect of modified prescription of Jin Gui Shen Qi wan on blood sugar is not significant by comparing with untreated diabetic control mice.But the urinary total protein excretion and the urinary total protein / creatinine ratio(UPCR)were significantly reduced in CM2 group.Besides,the renal hypertrophy index of mice had been improved as well.These results were similar to the results of ACEI group.The modified prescription of Jian Ling Decoction could lower blood glucose levels,reduce urinary total protein excretion,and improve renal hypertrophy index in mice,but there is no significant difference in urinary total protein / creatinine ratio.There was no significant difference between the CM1 group,CM2 group and ACEI group for the renal weight and renal hypertrophy index.Compared with DC group,CM1 group,CM2 group and ACEI group could reduce the expression of TGF-?1 protein and increase the expression of u PA protein,but there was no significant difference in IL-6 protein and PTEN protein between all groups.2.clinical trialsIn this study,the total number of patients who enrolled and completed the course of treatment was 38.11 of whom were male and 27 were female.After 12 weeks treatment of modified prescription of Jin Gui Shen Qi wan,the blood sugar levels before and after the treatment were(164.92 � 46.61)and(155.08 � 42.69)mg/d L with statistically significant difference;The serum creatinine levels before and after the treatment were(1.35 � 0.56)and(1.28 � 0.59)mg/d L with statistically significant difference;The urinary albumin levels before and after the treatment were(15.01 � 12.87)and(13.25 � 12.65)mg/d L with statistically significant difference;The TCM scores before and after the treatment were(30.79 � 8.71)and(20.05 � 6.55)mg/d L with statistically significant difference;The scores of quality of life before and after the treatment were(70.82 � 17.74)and(61.79 � 13.08)mg/d L with statistically significant difference.The total effective rate of TCM syndrome was 68.42% and the total effective rate of creatinine clearance rate and glomerular filtration rate were 55.26%.Conclusion:1.experimental models of STZ-induced diabetic nephropathy in miceAfter administration of modified prescription of Jian Ling Decoction,blood glucose levels,renal hypertrophy index,urinary excretion of total protein were decreased and renal pathological damage was relieved in STZ-induced diabetic mice.After administration of modified prescription of Jin Gui Shen Qi wan,urinary excretion of total protein,urinary protein/creatinine ratio and renal hypertrophy index were decreased and renal pathological damage was relieved in STZ-induced diabetic mice.The reno-protective effects had nothing to do with the improvement of blood glucose.All of three treatment groups including the modified prescription of Jian Ling Decoction,the modified prescription of Jin Gui Shen Qi wan and captopril could significantly improve urinary total protein excretion and renal hypertrophy index and relieved renal pathological damage in STZ-induced diabetic mice.The mechanism was related to the decrease of TGF-?1 and u PA protein in renal tissue,but not related to the expression of IL-6 and PTEN protein in renal tissue.2.clinical trialsEliminating dampness and phlegm and warmly invigorating spleen and kidney yang formulas like the modified prescription of Jin Gui Shen Qi wan could improve the clinical symptoms,fasting blood glucose,renal function,and quality of life in patients who were characterized with syndrome of Spleen Yang deficiency,Kidney Yang deficiency and Phlegm-damp stagnation.No severe adverse events were found during the treatment period,and the liver function were still in normal range.This study indicated that modified prescription of Jin Gui Shen Qi wan as a treatment for diabetic nephropathy was a safe and reliable drug.