Shenkang Injection protects against diabetic nephropathy in streptozotocin (STZ)-induced mice through enhancement of anti-oxidant and anti-inflammatory activities / 中草药·英文版

Objective: To investigate the protective effects and possible mechanisms of Shenkang Injection (SKI) on the diabetic nephropathy in streptozotocin-induced mice. Methods: STZ with the feeding of high fat diet (HFD) was used to induce diabetic mice. The balb/c mice and diabetic mice were then randomly divided into five groups: (1) control group, (2) model group, (3) alprostadil (Alp, 1.5 μg/kg) group, (4) SKI (30 ml/kg) group, (5) Alp (1.5 μg/kg) + SKI (15 ml/kg) group. After six weeks' treatment, blood, urine and kidney tissues were collected for biochemical assay, ELISA assay, and pathological analysis. Results: Diabetic mice exhibited evident manifestations of diabetic nephropathy (DN), as indicated by increased 24-h urine volume, urinary albumin and kidney weight index (P < 0.01), which could be attenuated by SKI treatment (P < 0.01). SKI was further found to improve abnormal morphology in glomerulus with increased glomerular volume and to decrease urinary N-acetyl-b-D-glucpsaminidase (NAG), β2-microglobulin (β2-MG), and kidney injury molecules-1 (KIM-1) levels (P < 0.05, P < 0.01). Plasma levels of anti-oxidant enzymes significantly reduced in the diabetic mice, and those decreases could be reversed by SKI and Alp treatments. Additionally, SKI obviously suppressed the diabetes-induced increases of pro-inflammatory cytokines (IL-6, IL-1β and TNF-α) (P < 0.01). Meanwhile, SKI was found to effectively attenuate the diabetes-induced coagulation dysfunction, as evidenced by lengthening prothrombin and thrombin time, and decreasing plasma levels of fibrinogen (FIB), 6-K-PGF1α and thromboxane B2 (TXB2) (P < 0.05, P < 0.01). With SKI and Alp combined treatment, the anti-oxidant activities and improvements of coagulation dysfunction were enhanced. Conclusion: SKI possesses a remarkable property to prevent diabetic nephropathy. The improvements of kidney function and hypercoagulability by SKI were enhanced with Alp combined treatment. The molecular mechanisms underlying the protection of SKI against DN may be related to enhancing the anti-oxidant and anti-inflammatory activities, and improving the coagulation dysfunction.

Preventive effect of Shenkang injection against high glucose-induced senescence of renal tubular cells / 医学前沿

Shenkang injection (SKI) is a classic prescription composed of Radix Astragali, rhubarb, Astragalus, Safflower, and Salvia. This treatment was approved by the State Food and Drug Administration of China in 1999 for treatment of chronic kidney diseases based on good efficacy and safety. This study aimed to investigate the protective effect of SKI against high glucose (HG)-induced renal tubular cell senescence and its underlying mechanism. Primary renal proximal tubule epithelial cells were cultured in (1) control medium (control group), medium containing 5 mmol/L glucose; (2) mannitol medium (mannitol group), medium containing 5 mmol/L glucose, and 25 mmol/L mannitol; (3) HG medium (HG group) containing 30 mmol/L glucose; (4) SKI treatment at high (200 mg/L), medium (100 mg/L), or low (50 mg/L) concentration in HG medium (HG + SKI group); or (5) 200 mg/L SKI treatment in control medium (control + SKI group) for 72 h. HG-induced senescent cells showed the emergence of senescence associated heterochromatin foci, up-regulation of P16 and cyclin D1, increased senescence-associated β-galactosidase activity, and elevated expression of membrane decoy receptor 2. SKI treatment potently prevented these changes in a dose-independent manner. SKI treatment prevented HG-induced up-regulation of pro-senescence molecule mammalian target of rapamycin and p66Shc and down-regulation of anti-senescence molecules klotho, sirt1, and peroxisome proliferator-activated receptor-g in renal tubular epithelial cells. SKI may be a novel strategy for protecting against HG-induced renal tubular cell senescence in treatment of diabetic nephropathy.

Protective effect and its mechanism of Shenkang injection on peritoneal mesothelial cells in CAPD mice / 天津医药

Objective To study the protective effect of Shenkang injection on peritoneal mesothelial cells (PMCs) of continuous ambulatory peritoneal dialysis (CAPD) mice, and explore the possible mechanism. Methods Forty ICR mice were randomly divided into blank control (A) group, peritoneal dialysis (B) group, low dose of Shenkang (C) group ( 2.5%dialysate+5 mL/kg Shenkang injection), medium (D) group (2.5% dialysate+10 mL/kg Shenkang injection) and high (E) group (2.5% dialysate+ 20 mL/kg Shenkang injection). Mice were observed for 4 weeks. Fasting blood glucose, total cholesterol, triglyceride and C-reactive protein (CRP) were detected by biochemical assay. Tumor necrosis factor-alpha (TNF-α), transforming growth factor-beta 1 (TGF-β1), vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF) levels of serum and dialysate were detected by ELISA. Pathological changes of peritoneal tissue were observed by HE staining. Expression and mRNA transcription levels of these four cytokines in the peritoneal tissue were detected by immunohistochemical staining and real-time PCR respectively. Results There were no significant differences in body weight, fasting blood glucose, total cholesterol and triglyceride between 5 groups of mice (P>0.05). Compared with group B, there was no significant difference in CRP level between group C and group E, but which was significantly decreased in group D (P<0.05). The serum and dialysate levels of TNF-α, TGF-β1, VEGF and CTGF were decreased in group C and group D. The serum and dialysate levels of TNF-αand TGF-β1 were significantly increased in group E (P<0.05), but there was no significant difference between VEGF and CTGF in group E. Compared with group E, except for CTGF in dialysate of group C, the serum and dialysate levels of TNF-α, TGF-β1, VEGF and CTGF were significant decreased in group C and group D (P<0.05). Damaged PMCs were found in group B, which were improved in various degrees in group C, group D and group E. Compared with group B, the protein expression and mRNA relative transcription levels of TNF-α, TGF-β1, VEGF and CTGF tended to decrease gradually in group C, group D and group E (P<0.05). Conclusion A certain concentration of Shenkang injection can protect PMCs by inhibiting the expression of TNF-α, TGF-β1, VEGF and CTGF in CAPD mice, so as to control the occurrence and development of peritoneal fibrosis.

Effect of Shenkang Injection on the Irbesartan Pharmacokinetics in Rats in vivo / 中国药房

OBJECTIVE:To study the effect of Shenkang injection on the irbesartan pharmacokinetics in rats in vivo. METH-ODS:18 SD rats were randomly divided into control group(normal saline)and Shenkang injection(calculated by crude drug as 4 g/kg),9 in each group,which were intraperitoneally injected twice a day,for 7 d. After 1 h of last administration,25 mg/kg irbe-sartan was intragastrically administrated. 0.3 mL sample blood was taken in fundus venous plexus before administration of irbesartan and after 0.25,0.5,1,2,4,8,12,24,32,48,56,72,96 h of administration. Using biphenyl diester as inner standard,HPLC was adopted to determine the plasma concentration of irbesartan in plasma of rats,and pharmacokinetic parameters were calculated by using non-compartmental model in Phoenix WinNolin? 6.1 pharmacokinetic software. RESULTS:After intragastrically adminis-trated irbesartan in rats in control group and Shenkang injection group,AUC0-96 h were (28.82 ± 10.49),(35.64 ± 9.99) mg·h/L;cmax were(0.64±0.15),(0.76±0.33)mg/L;tmax were(13.07±16.70),(10.23±3.97)h;CLZ/F were(0.85±0.35),(0.63±0.21) L/(h·kg);VZ/F were (38.24 ± 24.87),(30.99 ± 9.75) mL/kg respectively,with no statistical significances (P>0.05). CONCLU-SIONS:Shenkang injection will not affect the in vivo pharmacokinetics process of irbesartan in normal dose on rats.

Clinical efficacy of Shenkang injection combined with valsartan in the treatment of patients with early stage diabetic nephropathy / 中国基层医药

Objective To study the clinical effect of Shenkang injection combined with valsartan in the treat-ment of patients with early stage diabetic nephropathy,and its influence on the expression changes of homocysteine ( Hcy) ,transforming growth factor β1 ( TGF -β1 ) , the inflammatory cytokines high sensitive C -reactive protein (hs-CRP),and interleukin-6 (IL-6).Methods 138 cases of early stage diabetic nephropathy were randomly divided into control group ( conventional treatment, valsartan act as antihypertensive drug ) and treatment group ( treated with Shenkang injection combined with valsartan) by digital table.The total effective rate,glycemic index, renal function,levels of Hcy and TGF-β1,expression levels of inflammatory cytokines of two groups were compared. Results The total effective rate of the treatment group (91.30%) was significantly higher than the control group (76.81%) (χ2 =5.004,P<0.05).The values of FPG,2h PG,HbAlc,UAER,CysC and Scr were lower in the treat-ment group compared with the control group (t=4.514,3.743,4.754,6.214,3.925,3.777,all P<0.05).Com-pared with the control group,the concentrations of Hcy,TGF-β1,hs-CRP and IL-6 decreased significantly[ (11. 78 ±1.95)μmol/L vs (9.21 ±1.64)μmol/L,(71.32 ±14.88) mg/L vs (60.04 ±11.75) mg/L,(7.07 ±1.25) mg/L vs (5.81 ±1.14)mg/L,(50.24 ±21.86)pg/mL vs (32.55 ±17.01)pg/mL],the differences were statistical-ly significant (t=8.378,4.942,6.187,5.305,all P<0.05).Conclusion Shenkang injection combined with val-sartan for the treatment of early stage diabetic nephropathy has significant clinical efficacy.It can improve patients're-nal function and reduce blood sugar levels.The effects may be related with inflammatory response inhibition and Hcy, TGF-β1 levels reduction.

Effects and mechanisms of Shenkang injection promoting extracellular matrix degradation via regulating ERK1/2/MMPs signaling pathway in renal failure rats / 中国中药杂志

This study aimed to clarify preliminarily the effects and mechanisms of Shenkang injection (SKI) promoting extracellular matrix(ECM)degradation via regulating extracellular-signal regulated protein kinase(ERK)1/2/matrix metalloproteinases(MMPs)signaling pathway in renal failure rats. Twenty rats were randomly divided into 4 groups：the Sham group,the Model group,the SKI group and the Enalapril maleate(EM)group. The model rats with renal failure were induced by intragastric administration of adenine and unilateral ureteral obstruction(UUO). After modeling, the rats in SKI group and EM group were intervened by intraperitoneal injection of SKI or intragastric administration of the EM suspension,while the rats in Sham group and Model group were administrated with distilled water respectively for 3 weeks. The 24 h urinary protein excretion(Upro)and urinary N-acety1-β-D-glucosaminidase(UNAG)in all rats were tested after drug administration. All rats were sacrificed after drug administration for 3 weeks,blood and kidney were collected,renal morphological characteristics were observed. Furthermore,serum biochemical indices and the protein expressions of collagen type IV(CIV),MMP-2,MMP-9,tissue inhibitors of metalloproteinase(TIMP)-1,ERK1/2 and phosphorylated-ERK1/2(p-ERK1/2)in the kidney were evaluated respectively. The results indicated that,after the intervention of SKI,serum creatinine(Scr),blood urea nitrogen(BUN),uric acid(UA),albumin(Alb),Upro,UNAG and renal morphological change in model rats were improved at different levels,respectively. Moreover,these actions were similar to EM. In addition to these,SKI adjusted the protein expressions of MMP-2,MMP-9 and TIMP-1,and down-regulated the protein expressions of p-ERK1/2 in the kidney. Moreover,these actions were different from EM. In conclusion,SKI promotes ECM degradation and delays the progression of renal failure possibly through regulating ERK1/2 signaling pathway activation in the kidney and intervening MMPs/TIMP-1 expressions in vivo.

Stability of Shenkang Injection Respectively in Five Different Solutions / 中国药师

To explore the stability of Shenkang injection respectively in five different solutions to choose the best com-patibility program and improve the rational drug use. Methods:The pH value, insoluble particles and the content of protocatechuic al-dehyde in Shenkang injection were determined after mixed with five solutions. Results:In the 6-hour mixing, the change order of proto-catechuic aldehyde content was 0. 9% sodium chloride injection> 5% fructose injection> 10% invert sugar injection > 5% dextrose injection> 10% glucose injection, especially in 0. 9% sodium chloride injection, the content was decreased by 20%. The number of insoluble particles was increased after the 2-hour mixing except in 10% invert sugar injection solution, and the increase in the four so-lutions was beyond the requirement in the 2010 edition of Chinese Pharmacopoeia, and the number of insoluble particles in 0. 9% sodi-um chloride injection was the highest with the fastest change. During the experimental process, the pH value was stable. Conclusion:The five solutions show influence on stability of Shenkang injection in varying degrees. Shenkang injection in 0. 9% sodium chloride in-jection is the least stable, which should be used with caution in clinics, while invert sugar injection and glucose injection can be the best choice for Shenkang injection and the mixed solution should be used up in 2h.

Clinical Observation of Enalapril Folic Acid Tablet Combined with Shenkang Injection in the Treatment of Hypertensive Nephropathy / 中国药房

OBJECTIVE:To observe the clinical efficacy and safety of Enalapril folic acid tablet combined with Shenkang injec-tion in the treatment of hypertensive nephropathy. METHODS:90 patients with hypertensive nephropathy were randomly divided in-to control group and observation group. All patients were given anti-infection,regulating electrolyte balance,supplying amino acids and other conventional treatment. Based on it,control group was orally given Maleic acid enalaprilat folic acid tablet 1 tablet in the morning,once a day;observation group was additionally given 100 ml Shenkang injection adding into 300 ml 10% Glucose injec-tion by intravenous infusion,20-30 drops/min,once a day. The treatment course for both groups was 4 weeks. The clinical effica-cy,and clearance rate of creatinine(Ccr),serum creatinine (Scr),morning urine test urine protein (Up)/urine creatinine (Ucr), serum malondialdehyde(MDA),superoxide dismutase(SOD),total antioxidant capacity(T-AOC)before and after treatment and incidence of adverse reactions in 2 groups were observed. RESULTS:The total effective rate in observation group was significantly higher than control group,the difference was statistically significant(P0.05). And there was no significant difference in the incidence of adverse reactions(P>0.05). CONCLUSIONS:Based on the conventional treat-ment,Enalapril folic acid tablet combined with Shenkang injection has better efficacy than only Enalapril folic acid tablet in the treatment of hypertensive nephropathy,with similar safety.

Influence of Shengkang injection with Irbesartan on trace albumin excretion and blood rheology in early diabetic nephropathy / 中国综合临床

Objective To observe the influence of Shenkang injection with Irbesartan on urinary albumin excretion rate(UAER)in 24 hours,serum lipid and blood rheology in early diabetic nephropathy.Methods One hundred and fifty normotensive early diabetic nephmpathy were randomly divided into control group,Shenkang injection group and Shenkang injection with Irbesartan group.Three groups were treated with diabetic education,dietary regimen,suitable physical activity and oral anti-hyperglycemic agents till their fasting blood glucose(FBG)was less than 7.0 mmol/L and postprandial blood glucose(PBG)was less than 10.0 mmol/L.The control group was treated with oral anti-hyperglycemic agents or/and insulin.Shenkang injection group was additionally treated with Shenkang solution.Shenkang solution was mixed by Shenkang injection 100 ml,5% glucose 250 ml and insulin 2-3 U.Shenkang solution was intravenously guttaed once a day.Shenkang injection with Irbesartan group was additionally treated with Irbesartan 0.15 g,once a day,taken orally.A course was taken for 3 weeks.The levels of UAER,serum lipid and blood rheology were detected immediately and 9 weeks after treatment.Results In the Shenkang injection group,compared with those before treatment,them were significant difference in UAER (87.44 ±10.06)μg/min vs.(116.55 ± 33.42)μg/min),serum lipid( (4.22 ± 0.70)mmol/L vs.(4.88 ± 0.69 )mmol/L)and blood rheology( (5.77 ±0.53)mPa·s vs.(7.38 ± 0.41)mPa·s)(P ＜ 0.05).In the Shenkang injection with Irbesartan group,compared to those before treatment,there were also significant difference in UAER ( (61.90 ±28.02)μg/min vs.(123.37 ± 29.98)μg/min),serum lipid( (4.00 ± 0.14)mmol/L vs.(4.90 ± 0.12)mmol/L)and blood rheology( (5.11 ±0.41)mPa·s vs.(7.27 ± 0.44)mpa·s)(P ＜ 0.05).Whearas in the control group,no significant difference were found compared with those before treatment.Furthermore,the improvements in the Shenkang injection with Irbesartan group were better than those in the Shenkang injection group(P ＜ 0.05).Conclusions Shenkang injection can reduce UAER,decrease serum lipid and improve renal blood rheology.Iteffectively prevents diabetic nephropathy in earlier period and maybe have synergy with Irbesartan.

Observation of Curative Efficacy of Shenkang Injection for Chronic Renal Failure / 中国药房

OBJECTIVE:To observe the clinical efficacy of Shenkang injection,a new compound preparation for chronic renal failure(CRF).METHODS:A total of 84 CRF patients were randomly divided into two groups:the treatment group(n=54)was given routine antihypertensive therapy and infused(iv gtt)with Shenkang injection 60～ 100mL(dissolved in 10% GS 300mL or NS 300mL)q.d for 30d;the control group(n=30)was infused(iv gtt)with Compound danshen injection 250mL q.d for 30d.RESULTS:As compared with the control group,serum levels of creatinine and urea nitrogen(UN)in treatment group had a greater reduction(P