Effects Of Yiqi-Yangyin-Xiaozheng-Tongluo Of Traditional Chinese Medicine On Slit Diaphragm Proteins Of Hige Glucose Cultivated Podocyte

Objective: Diabetic nephropathy has become one of the leading cause of end-stage kidney disease, the occurrence of a large amount of proteinuria is typical signs of diabetes renal tissue damage, podocyte damage can lead to loss of glomerular filtration membrane integrity and a large number of protein leakage, podocyte plays an important role in proteinuria..In previous research, we with qi and nourishing Yin disease know luo for the treatment of DN solution, has confirmed that this method can pass the DN rat TGF-β/Smads, oxidative stress and the influence of the RASS system play a role of treatment of DN.This study following the mice podocyte as the research object, from the level of cell to observe the expression for podocyte slit diaphragm protein- nephrin and podocin, CD2 AP on the method of yi qi xiao zheng tong luo.Methods:1 The preparation for yiqi-yangyin-xiaozheng-tongluo of traditional Chinese medicine serum and normal serum12 healthy male SD rats, SPF, weight 200-220 g, provided by experim- ental animal center of hebei medical university, animal certificate No. 1404079.The animals were randomly divided into 3 groups, the normal group, low dose group, middle dose serum drug group, high-dose serum drug group.Normal group was given the corresponding dose of distilled water, the rest three groups at a low dose group, according to the formula Rubner Meeh for clinical half quantity for clinical dosage, middle dose group, high dose group is two times of clinical dosage convert lavage doses, Chinese medicine used for the extraction of traditional Chinese medicine granules, purchased from hebei medical university affiliated hospital of TCM pharmacy in hebei province.Between groups each 2 ml, lavage, dosing interval of 12 hours at a time, at the end of the 3d after consecutive gastric cardiac blood after 2h, 4500 r/min centrifuge for 10 min to separate serum, 56℃ constant temperature water bath live for 30 min, 0.22 um microporous membrane filtration,-20℃ refrigerated. Set aside.2 The culture of mice podocyte and the influence of nephrin and podocin, CD2 AP for yiqi-yangyin-xiaozheng-tongluo of traditional Chinese medicine serumConditional immortalized mouse cells in 33℃, gamma-under the licensing conditions for the existence of IFN training after the wedding, at 37℃, the absence of gamma-IFN subculture 10-14 days, promote cell differentiation and mature.Differentiation of mature enough cells were randomly divided into 6 groups: Normal Control group(Normal Control, NC), High glucose, the Control group(High Glucose, HG) and Normal Rat serum in the Control group(Normal Rat, NR) group, Low dose drug serum(Low dosed of drug, LD), the dose of drug serum group(Normal dosed of drug, ND) group, High dose of drug serum(High dosed of drug, HD), NC group cells give culture medium containing D- Glucose tendency for 5.5 L+5% fetal bovine serum, HG group was given culture medium containing 30 tendency/L +D-Glucose 5% fetal bovine serum, NR group was given culture medium containing 30 tendency/L+D-Glucose 5% fetal bovine serum+5% to Normal Rat serum, LD group was given culture medium containing 30 tendency/L+D-Glucose 5% fetal bovine serum+5% serum group, Low dose of Chinese traditional medicine ND group was given culture medium containing 30 tendency/L+D-Glucose 5% fetal bovine serum+5% dose of Chinese traditional medicine serum group, HD group was given culture medium containing 30 tendency/L+D-Glucose 5% fetal bovine serum+5% High dose of Chinese traditional medicine serum group.Each training stimulus after 24 h, collect enough cells application immunochemical method, western blotting, Real- time PCR detection of nephrin and podocin, CD2 AP gene and protein expression.Results:NC group compared with HG group, NC group of nephrin and podocin, CD2 AP gene expression and protein content were increased significantly in the HG group(P<0.01); HG group compared with NR group, two groups of nephrin and podocin, CD2 AP gene expression and protein content were no statistical significance(P>0.05); Treatment group(LD, ND, HD) compared with HG group, treatment group nephrin and podocin, CD2 AP gene expression and protein content than HG increased(P<0.05), and the amount of nephrin and podocin, CD2 AP gene expression and protein content and drug serum used doses were positively correlated.Conclusions:1 The expression of nephrin and podocin, CD2 AP on the high glucose cultivated mice podocyte was decreased.2 Yiqi-yangyin-xiaozheng-tongluo of traditional Chinese medicine can significantly raise nephrin and podocin and CD2 AP protein and m RNA expression of high glucose mice podocyte.Yiqi-yangyin-xiaozheng-tongluo of traditional Chinese medicine can through raising nephrin and podocin and CD2 AP protein and m RNA expression treat diabetic nephropathy.