| Objective:This study was aimed to investigate the effect and mechanism of Mahuang Formula on 5/6 nephrectomy chronic kidney disease in rats.Methods:100 sprague dawley(SD)rats were randomly distributed into seven groups,which were normal group(Nor),sham operation group(Sham),model group(Mod),low dosage group(Low),medium dosage group(Med),high dosage group(Hig)and benazepril group(Bel).After the 5/6 nephrectomy model was successfully established,the rats in the Low,Med and Hig group were administrated with Mahuang Formula for 63consecutive days.The rats in the benazepril group were administrated benazepril,meanwhile,the rats was intragastricly RO water in the model group and normal group,sham group rats for nine weeks.The blood sample,urine sample and kidneys were harvested as previously at.10th week,13th week,16th week.The level of serum creatinine,blood urea nitrogen and 24-hour urinary protein were tested by automatic biochemical analyzer.And at 16~thh week,the rats were sacrificed and the kidney specimens were collected and saved in refrigerator for use.To observe renal pathologic injury and fibrosis was performed HE and Masson staining of kidney.The expression of FN,α-SMA of kidney tissue were assessed by immunohistological staining,in addition,the expression of FN,TGF-β1,P-smad3,smad7 in kidney tissues were assessed by western blot.Results:(1)The index of kidney function:the serum creatinine,blood urea nitrogen,and 24-hour urine protein of Normal,sham group show no obvious changes over time,and the serum creatinine,blood urea nitrogen,the 24-hour urinary protein was significantly increased in all time points(P<0.05).At 10 week,compared with model group,the BUN,Scr and 24h UP in CKD significantly decreased.As compared to model group,the BUN,Scr and 24h UP in high dose group and benazepril group signicantly decreased(P<0.01).Further,compared with medium dose group and low dose group,the 24h UP in high dose group and benazepril group signicantly decreased(P<0.05).At13 week,compared with model group,the BUN in CKD significantly decreased.But,there was no significance.As compared to low dose group,the BUN in Med,high dose group and benazepril group significantly decreased(P<0.05).Compared with model group,the Scr in CKD significantly decreased.And the BUN in Med,high dose groupandbenazeprilgroupsignicantlydecreased(P<0.05).Further,compared with model group,the 24h UP in CKD decreased.In addion,the 24h UP in high dose group and benazepril group significantly decreased(P<0.05).As compared to low dose group,the 24h UP in high dose group and benazepril group significantly decreased(P<0.05).At 16 week,compared with model group,the BUN,Scr and 24h UP in CKD significantly decreased.As compared to model group,the Scr in high dose group and benazepril group signicantly decreased(P<0.05).Further,the BUN in high dose group significantly decreased(P<0.05).While the 24h UP in Med,high and benazepril group significantly decreased(P<0.05).As compared to low dose group,the 24h UP in high dose groupandbenazeprilgroupsignificantlydecreased(P<0.05).Compared with medium dose group,the 24h UP in high dose group decreased(P<0.05).(2)Pathological structure of kidney:At 16 week after molding,HE and Masson staining of Normal group,sham group showed no change,and HE and Masson staining in the kidneys of rats with CKD revealed that glomerular structure sclerosis,shrinkage,the scrotal adhesion,significant renal tubular expansion,tubular tube type,obvious infiltration of various inflammatorycells,fibrosisinrenalinterstitiumhad occurred.Pathological Changes were substantially attenuated following Mahuang Formula and benazepril treatment.In addition,Mahuang Formula administration,at a dose of 5ml/kg body weight,ameliorates kidney injury obviously.(3)Renal fibrosis Index:At 16th week,compared with sham group,the IHC of FN andα-SMA showed that the expression of FN andα-SMA in 5/6nephrectomy rats was significantly increased(P<0.05).And in comparison with model group,the expression ofα-SMA and FN in the kidney tissues in medium dose group,high dose group and benazepril group rats reduced in different degrees(P<0.05),and compared with medium dose group,the expression of FN andα-SMA in high dose group decreased.5/6 nephrectomy significantly induced disease in rats.Chronic injury significantly upregulatedα-SMA,TGF-β1 and P-smad3,decreased smad7 expression in the kidney as compared to sham group.Concomitantly,Mahuang Formula administration and benazepril treatment significantly downregulatedα-SMA,TGF-β1 and P-smad3,decreased smad7expression in the kidney of rats as compared to model group.However,high dosage group significantly increased smad7expression.Conclusions:These results reveal the therapeutic effects of Mahuang Formula against kidney injury in rats through closely modulating with TGF-β1/smads signaling pathway. |
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