Effect Of "JianPiBuShenHuoXue" Therapy On PAI-1in The Diabetic Rats’ Kidney

Objective: Jianpibushenhuoxuefang (It is composed of huangqi,dangshen, baizhu, shanyao, shanzhuyu, nvzhenzi, hanliancao, xianmao,xianlingpi, chuanxiong, fuling, honghua, danggui, tusizi, chenpi, muxiang,zexie) has efficacy of jianpibushen, huoxuehuayu. After years of clinicalpractice, Mentor Professor Zhang Gengliang found that jianpibushenhuoxuetherapy can play a prevention and treatment effect on diabetic renal disease inearly stage of diabetes. In this experiment, through the establishment ofdiabetic rats model by Intraperitoneal injecting streptozotocin(STZ), andexerting medicine intervention, to observe the effect of jianpibushenhuoxuetherapy on blood glucose, blood lipid, renal function, twenty-four hours’urinary albumin excretion rate, renal pathological morphological changes andthe expression of plasminogen activator inhibitor-1(PAI-1) in diabetic rats,then preliminary to discuss this theraphy’s mechanism of protecting renaltissues.Methods: Forty-six clean grade healthy of Wistar rats(male, weightfrom160g to180g)were selected. After one week’s adaptive feed by ordinaryfeeds, select ten rats randomly according to the weight as the normal controlgroup, the remaining rats were used for copying the diabetes model, injectedwith streptozotocin (STZ) in abdominal cavity, which was dissolved into1%buffer solution of citric acid of STZ according to fifty-five mg/kg, the normalgroup were injected with the corresponding1%citric acid-sodium citrate.After seventy-tow hours later, to test blood glucose from the tail vein. Weselect the rats which urine and water intake increased significantly and thenumerical value of the blood glucose more than16.7mmol/L as a successdiabetic rats model.The diabetic rats were randomly divided into model control group, jianpibushenhuoxuefang group (called traditional Chinese medicine group forshort), Irbesartan group (called Western medicine group for short), and twelverats in each group. After three days of the success modeling, the drugs startedbe given to the rats according to adult dosage10times, once a day. The rats ofChinese medicine group were given the jianpibushenhuoxuefang eleven ml/kgweight per day (equivalent to3.5g/ml herbs) by gavage, the Western medicinegroup were given the irbesartan five ml/kg weight per day (equivalent to7.5mg/ml simvastatin) by gavage, the normal group and model group weregiven the corresponding quantity’s physiological saline by gavage. Each groupwas given the medicine continuously for eight weeks by gavage. On the daybefore their sacrificed, the rats were placed in the metabolic cages to collect24-hour urine, noted urine volume, and measured urinary albumin excretionrate. The rats were forbidden to eat but not water for twelve hours after the lastgavage. All rats were anesthetized by injecting2%napental twenty mg/kg intoabdomen, taking blood from femoral artery to test the blood glucose, totalcholesterol (TC), triglycerides (TG), blood urea nitrogen (BUN) and serumcreatinine (Scr). Rats were killed, then the right kidney was taken out, itscapsule was removed, blood was siphoned by filter paper. After weighting,kidney’s weight was recorded and the hypertrophy index (kidney weight/bodyweight, KW/BW) was calculated. Part of the renal tissues were paraffinembedded and conventional biopsied, then were lined HE staining under lightmicroscope to observe the morphological changes of renal pathology. At thesame time, some other renal tissues were taken out as specimens by asepticoperation, fixed in4％formaldehyde, using immunohistochemical method todetect the expression of plasminogen activator inhibitor-1(PAI-1).Results:1General situation of all the ratsGeneral situation included the rats diets, water quantity, urine output,weight, mental state, fur, and the status of the activity. The normal rats havenormal diet, normal drinking, normal urine, weight gain significantly, havesmooth surfaces and act freely. But the modle group showed more drink, more food, polyuria, weight loss obviously，apathetic and shape gradually thinner,color dry-taek and messy, the reaction retardation. Over Time some evenappeared a variety of concurrent symptoms of diabetes, such as hemiplegica,cataract, limb edema and so on. The performance of treatment groupsalleviated than that of the model group. There was one death in each group ofmodel group and Western medicine group. The eventually death rats numberof model group, traditional Chinese medicine group, Western medicine groupwas one, two and one. The main cause of the death was gavage, tearing ateach other and infection，hemiplegica and starvation.2The blood glucose of the rats in each groupComPared with the normal group, the blood glucose of the model groupand two treatment groups was significantly higher. The difference wasstatistically significant (P＜0.01). The blood glucose of the traditionalChinese medicine group was significantly lower than model group. Thedifferences are significant (P＜0.01). But the blood glucose of model groupand Western medicine group was not of significant differences (P＞0.05).3The blood lipid changes of each groupThe TC and TG of model group, traditional Chinese medicine group andWestern medicine group were significantly higher than that of normal group(P＜0.01). Traditional Chinese medicine group compared with the modelgroup decreased significantly (P＜0.01). The blood lipid of Western medicinegroup is reduced than that of model group, but no significant difference (P＞0.05).4The renal function changes of each groupThe BUN and Scr of model group, traditional Chinese medicine groupand Western medicine group were significantly higher than that of normalgroup (P＜0.01). Compared with model group, both of the treatment groupcan reduce BUN and Scr. The difference was statistically significant (P＜0.01). And the renal function indexes of traditional Chinese medicine andwestern medicine group had no significant difference (P＞0.05).5The24h urinary albumin excretion rate changes of each group Compared with normal group,24h urinary albumin excretion rate ofmodel group increased significantly (P＜0.01). Traditional Chinese medicinegroup’s24h urinary albumin excretion rate was significantly lower than modelgroup’s with statistical significance (P＜0.01), without statistical significancecompared to western medicine group (P＞0.05).6The hypertrophy index (kidney weight/body weight) changes of each groupThe kidney weight/body weight ratio of model group was obviouslyhigher than that of normal group (P＜0.01). The kidney weight/body weightratios of Irbesartan group and jianpibushenhuoxuefang group are significantlylower than that of model group, having significant difference compared withmodel group (P＜0.01). And between the traditional Chinese medicine groupand western medicine group there was no significant difference (P＞0.05).7Effect on experimental diabetic rats’ renal pathological morphologyIn normal group，renal glomerulus was integrity, no increase in renalglomerulus and cell number, no significant abnormalities in glomerularcaPillary basement membrane and mesangial matrix. In model group, renalglomerulus and nucleus number were increased, glomerular capillarybasement membrane thickened, mesangial cell mild proliferation, mesangiumwidened. Proximal epithelia cell swelled in renal tubular. Some partial tubularepithelial cells had a bulb-like degeneration. The rats’ kidney lesions oftraditional Chinese medicine group and Western medicine group significantlyimproved compared with model group.8Effect on the expression of the plasminogen activator inhibitor-1(PAI-1) inthe diabetic rats’ kidneyComPared with normal group, the expression of the plasminogenactivator inhibitor-1(PAI-1) in the kidney of model and two treatment groups’rats was significantly higher than the normal group (P＜0.01). After drugtreatment, the treatment group compared with the model group, the expressionof PAI-1was significantly reduced (P＜0.01), the difference was statisticallysignificant. The traditional Chinese medicine group compared with thewestern medicine group had no significant difference (P＞0.05).It’s suggested that irbesartan and jianpibushenhuoxuefang’s protective effect on diabetickidney can be possibly realized through down-regulated the expression ofplasminogen activator inhibitor-1(PAI-1).Conclusions:1Jianpibushenhuoxuefang can improve the general observation ofbehavior of diabetic, lower the blood glucose, blood lipid, serum creatinineand blood urea nitrogen, reduce urine protein, then protect kidney.2Jianpibushenhuoxuefang had the similar influence of diabetic ratswith irbesartan. Among this, jianpibushenhuoxuefang was better thanirbesartan in the hypoglycemic and lowering blood lipid.3Jianpibushenhuoxuefang can reduce the expression levels ofplasminogen activator inhibitor-1(PAI-1), provided a new way and newthought for the research of traditional Chinese medicine to protect kidneymechanism.