| Objective:Diabetic nephropathy(DN) is one of the most hazardous complications of diabetes.It is also the main reason that can cause diabetes to death.Renal hypertrophy in the early stage is the characteristic of diabetic nephropathy (DN).With the progress of diabetic nephropathy,a complex of pathological changes,including glomerulosclerosis,tubular atrophy and inter- stitial fibrosis.For the past few years following the improvement of molecular biology,people discover that apoptosis partici- pates the DN's mechanisms.Cell apoptosis is regulated by various factors.Bcl-2 and Bax are gene family associated with apoptosis,it makes an important role on cell apoptosis.We use Erhuangtangshenkang to heal DN and get fine curative effect.We reproduce DN model rats,observe rats'blood sugar,renal function,cell cycle,apoptosis and associated protein Bcl-2 and Bax,approach the cure mechanism,so as to find an effective way for the prevention and cure of diabetic nephropathy (DN).Methods:Ten rats were selected for sham-operation group(SO)randomly in the sixty Sprague-Dawley rats.Others were had right nephrectomy.We injected the rats streptozo cin(STZ) intraperitoneally after 2 weeks referinig to 50mg/kg.We regarded the rats as model rats whose blood sugar≥16.7mmol/L,urine glucose are from +++~++++and urine protein were masculine.The model rats were randomly divided into: Model group(M),Benazeprill group(B),Low-dose Erhuang tangshenkang group(EL),high-dose Erhuangtangshenkang group (EH),n=10.Animals were sacrificed after 8 weeks,their blood was collected and tested Scr,BUN,GLU,INS.The left kidney was removed,peel the erwelope away, weighed, and calculate the ratio of kidney weight and body weight (KW/BW).The other parts of left kidneys was harvested for pathological study.TUNEL is used for observing apoptosis cells.Flow cytometry is used for observing apoptosis cells,cell cycle,Bcl-2 and Bax.Immunehistochemistry method is used for investigate the expression of Bcl-2 and Bax.Results:1 The effect of Erhuangtangshenkang on BUN,ScrBUN of Group M were raised obviously compared to the group of SO(p<0.01).The treatment groups was lower than that of group M(p<0.01).The effect between EH EL group and B group have distinct difference(p<0.01).There were no distinct differences among EH and EL groups(P>0.05).Scr of Group M were raised obviously compared to the group of SO(p<0.01).The treatment groups was lower than that of group M(p<0.01).There were no distinct differences among the treatment groups(P>0.05). 2 The effect of Erhuangtangshenkang on INS,GLUINS of Group M were raised obviously compared to the group of SO(p<0.01).The treatment groups was lower than that of group M(p<0.01).The effect between EH group and B group has distinct difference(p<0.01).There were no distinct differ- ences among EH and EL groups(P>0.05).GLU of Group M were raised obviously compared to the group of SO(p<0.01).The treatment groups was lower than that of group M(p<0.01).The EL group was lower than B group (p<0.05).The effect between EH group and B group has distinct difference(p<0.01).There were no distinct differences among EH and EL groups(P>0.05).3 The effect of Erhuangtangshenkang on KW/BWKW/BW of Group M were raised obviously compared to the group of SO(p<0.01).The treatment groups was lower than that of group M(p<0.01).The EH group was lower than B group (p<0.05).There were no distinct differences among EH and EL groups(P>0.05).4 Detecting apoptosis in renal cortex by flow cytometry4.1 Detecting cell cycle by flow cytometryAt the stage G0/G1,the number of group M decreased obviously compared with group SO(p<0.01),The group B is higher than group M(p<0.05),The group EH,EL were increased obviously comparing with group M(p<0.01).The EH group was higher than B group(p<0.05).There had the therapeutic equivalence compared with group EL and EH(p>0.05).The effect between group B and group SO was different(p<0.05).At the stage S,the number of group M increased obviously compared with group SO(p<0.01).The group B is lower than group M(p<0.05).The group EH,EL were decreased obviously comparing with group M(p<0.01).The EH group was lower than B group(p<0.05).There had the therapeutic equivalence compared with group EL and EH(p>0.05).The effect between group B and group SO was different(p<0.05).At the stage G2M,every groups had no statistical signi- ficance(p>0.05).The value of PI of group M increased obviously com- pared with group SO(p<0.01).The group B is lower than group M(p<0.05).The group EH,EL were decreased obviously comparing with group M(p<0.01).The EH group was lower than B group(p<0.05).There had the therapeutic equivalence compared with group EL and EH(p>0.05).The effect between group B and group SO was different(p<0.05).4.2 Detecting apoptosis rate by flow cytometryThe apoptosis rate of group M increased comparing with all groups(p<0.01).The group B is higher than group SO (p<0.01).The effect between group EL and group SO was different(p<0.05).There had the therapeutic equivalence compared with EL and EH groups(p>0.05).4.3 Detecting the relative amount of Bcl-2 and Bax by flow cytometryThe relative amount of Bcl-2 of group M was lower than group SO(p<0.01).The group B is higher than group M (p<0.05).The effect between EH EL group and M group have distinct difference(p<0.01).There were no distinct differences among the treatment groups(P>0.05).The relative amount of Bax of group M was higher than group SO(p<0.01).The group B is higher than group M (p<0.05).The effect between EH,EL group and M group have distinct difference(p<0.01).There were no distinct differences among the treatment groups(P>0.05).5 Detecting the relative amount of Bcl-2 and Bax by Immune- histochemistry methodThe level of gene expression of Bcl-2 in group M was decreased compared with group SO(p<0.01).The group B is higher than group M (p<0.05).The effect between group EH,EL and group M have distinct difference(p<0.01).There were no distinct differences among the treatment groups(P>0.05).The level of gene expression of Bax in group M was increased compared with group SO(p<0.01).The group B is lower than group M (p<0.05).The effect between group EH,EL and group M have distinct difference(p<0.01).There were no distinct differences among the treatment groups(P>0.05).6 Detecting apoptosis rate by TUNELWe found little apoptosis cells in glomeruli of group SO. Each treatment groups apoptosis rate were lower than that of group M (p<0.01).There were no distinct differences among the treatment groups(P>0.05). The apoptosis cell in nephric tubule in group M was higher than all groups (p<0.01).The effect between group EH and group B have distinct difference(p<0.05).There had the therap- eutic equivalence compared with EL and EH groups(p>0.05).Conclusion:1 Erhuangtangshenkang can decrease the level of BUN,Scr,GLU,INS in the blood.2 Erhuangtangshenkang can decrease the number of kidney apoptosis.3 Erhuangtangshenkang can interfere in the stage of G0/G1 and S,decrease PI.So it can decrease the excessively cell generation.4 Erhuangtangshenkang can grow downwards the relative amount of Bcl-2 and Bax.It can interfere in kidney apoptosis.
|
PDF Full Text Request