Therapeutic mechanism of Shenbing Decoction Ⅲ for renal fibrosis in chronic kidney disease: a study with network pharmacology, molecular docking and validation in rats / 南方医科大学学报

OBJECTIVE@#To observe the effect of Shenbing Decoction Ⅲ for improving renal function and pathology in rats with 5/6 nephrectomy and analyze its therapeutic mechanism for renal fibrosis in chronic kidney disease using network pharmacology combined with molecular docking.@*METHODS@#Forty male SD rats were randomized into two groups to receive two-staged 5/6 nephrectomy (n=30) or sham operation (n=10), and 2 weeks after the final operation, serum creatinine level of the rats was measured. The rats with nephrectomy were further randomized into Shenbing Decoction Ⅲ group, losartan group and model group for daily treatment with the corresponding drugs via gavage starting at 1 week after 5/6 nephrectomy. After 16 weeks of treatment, serum creatinine and urea nitrogen levels of the rats were measured, and HE staining and Western blotting were used to examine the changes in renal pathology and fibrosis-related factors. Network pharmacology combined with molecular docking study was performed to explore the therapeutic mechanism Shenbing Decoction Ⅲ against renal fibrosis in chronic kidney disease, and Western blotting was used to verify the expressions of the core targets.@*RESULTS@#Compared with those in the model group, the rats receiving 5/6 nephrectomy and Shenbing Decoction Ⅲ treatment showed significantly reduced serum creatinine and urea nitrogen levels, lessened renal pathologies, and improvement of the changes in epithelial mesenchymal transition-related proteins. Network pharmacological analysis showed that the main active ingredients of Shenbing Decoction Ⅲ were acacetin, apigenin, eupatilin, quercetin, kaempferol and luteolin, and the key targets included STAT3, SRC, CTNNB1, PIK3R1 and AKT1. Molecular docking study revealed that the active ingredients of Shenbing Decoction Ⅲ had good binding activity to the key targets. Western blotting showed that in rats with 5/6 nephrectomy, treatment with Shenbing Decoction Ⅲ obviously restored the protein expression of STAT3, PI3K, and AKT in renal tissue.@*CONCLUSION@#Shenbing Decoction Ⅲ can reduce renal injury induced by 5/6 nephrectomy in rats, and its therapeutic effects are mediated possibly by its main pharmacologically active ingredients that alleviate renal fibrosis via modulating multiple targets including STAT3, PIK3R1, and AKT1.

Effects of warming yang-dispelling turbidity-dredging collaterals method on renal interstitial fibrosis in rats with chronic renal failure and mechanisms involved / 中草药

Objective To investigate the effects of warming yang-dispelling turbidity-dredging collaterals method on renal interstitial fibrosis in rats with chronic renal failure caused by 5/6 nephrectomy and explore its mechanisms. Methods Eight rats were randomly chosen as sham operation group (Sham), while the remaining 24 rats underwent 5/6 nephrectomy were divided into model group (5/6Nx), warming yang-dispelling turbidity-dredging collaterals method group (WDD) and Niaoduqing group (NDQ), and the rats in each group were administrated with corresponding drugs. After the intervention of 12 weeks, all the rats were sacrificed and the kidneys were then removed. HE, PAS Masson, Picrosirius red staining were conducted to assess kidney pathological changes and renal fibrosis. The protein expression of Collagen was tested by both immunofluorescence and Western blotting, and the mRNA expression was evaluated using qRT-PCR. The expression of protein related with the process of EMT, including α-SMA, E-Cadherin and Vimentin, were tested with immunohistochemistry and Western blotting. Results Compared with the model group, warming yang-dispelling method alleviated kidney pathological injury and renal fibrosis, and significantly down-regulated the protein and mRNA expression of Collagen . Moreover, warming yang-dispelling turbidity-dredging collaterals method regulated the process of EMT by decreasing the high expression of α-SMA and Vimentin, and increasing the low expression of E-Cadherin. Conclusion Warming yang-dispelling turbidity-dredging collaterals method can effectively attenuate renal fibrosis and injury and its underlying mechanisms may be related with its inhibitory effects on Collagen expression and regulation of EMT.

Effect of Kangxianling Decoction on TGF-β1/Smad Signal Pathway in Renal Fibrosis Rats / 中国实验方剂学杂志

Objective: To study the therapeutic effect of Kangxianling Decoction on renal fibrosis induced by 5/6 nephrectomization in rats, and discuss its action mechanism. Method: Totally 50 SD rats were randomly divided into normal control group (n=10), sham-operation group (n=10), and 5/6 nephrectomized renal fibrosis model group (n=30). After successful modeling, rats were randomly divided into the model group, Kangxianling group, and Losartan Potassium group. Rats in the Losartan Potassium group were administered with Losartan Potassium by gastrogavage; rats in the Kangxianling group were administered with Kangxianling by gastrogavage. Equal volume of distilled water was administered to rats in the other groups. Rats were sacrificed after 8 weeks of consecutive medication, and then serum creatinine (SCr), urea nitrogen (BUN), 24 hour urine protein (24 h-Pro) were measured in each group. Hematoxylin-eosin (HE) staining was used to observe the renal pathological changes and the score of Kidney damage was made. Masson staining was used to observe the degree of renal fibrosis. Western blot and real-time fluorescent quantitative polymerase chain reaction(Real-time PCR) were used to detect the protein and mRNA expression levels of transforming growth factor (TGF-β1), Smad2, Smad3, and Smad7 in kidneys. Result: As compared with the normal control and sham-operation group, the renal tissue fibrosis, score of kidney damage, SCr, BUN and 24 h-Pro levels were higher in model group (Pβ1, Smad2, and Smad3 (PPPβ1, Smad2, Smad3 were reduced (PPConclusion: Kangxianling decoction could alleviate renal fibrosis by inhibiting TGF-β1/Smad signal pathway.

Effect of Kangxianling Decoction on Renal Fibrosis and ACE-AngⅡ-AT1R Axis in 5/6 Nephrectomized Rats / 中国实验方剂学杂志

Objective: To study the therapeutic effect of Kangxianling decoction on renal fibrosis induced by 5/6 nephrectomy, and angiotensin converting enzyme-angiotensin Ⅱ-angiotensin Ⅱ 1 receptor (ACE-AngⅡ-AT1R) axis. Method: Totally 50 SD rats were randomly divided into the following groups:control group (n=10), sham-operation group (n=10), 5/6 nephrectomized renal fibrosis model group (n=30). After two weeks, the rats in operation group were divided into the model group, Kangxianling group, and losartan potassium group, n=10 in each group. Rats in losartan potassium group were administered with losartan potassium by gastrogavage, and rats in Kangxianling group were administered with Kangxianling by gastrogavage. Equal volume of saline was administered to rats in the other groups. The rats were put to death after 16 weeks of consecutive medication, and serum creatinine(SCr), blood urea nitrogen(BUN), 24 h urine protein(24 h-Pro) were measured in each group. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of kidney tissues, and the degree of renal fibrosis was observed by Masson staining. The expressions of ACE1 and AT1R were detected by immunohistochemistry. The protein expression levels of ACE1, AngⅡ and AT1R were determined by Western blot. Result: Compared with control and sham-operation groups, SCr, BUN and 24 h-Pro in model group were significantly increased (PPPPConclusion: Kangxianling decoction can delay the progress of renal fibrosis in 5/6 nephrectomized rats, which is closely related to the inhibition of ACE-AngⅡ-AT1R axis activation.

Progress in Research of Kangxianling Decoction in Delaying Renal Fibrosis / 中国实验方剂学杂志

Renal fibrosis is a common pathological change in the later stages of all kidney diseases. It is a multi-cytokine, multi-signal pathway, multi-factor driven chronic kidney disease. It includes renal interstitial fibrosis, tubular sclerosis and glomerular sclerosis, which eventually leads to chronic renal failure. From health through injury to loss of function, the disease process is closely related to the degree of deterioration of renal function and the prognosis of chronic kidney disease. There is no effective western medicine for the treatment of renal fibrosis. Professor HE Liqun from Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine has summarized the Kangxianling decoction through decades of long-term practical experience. It has the effects in strengthening the spleen and replenishing Qi, clearing away dampness and heat, promoting blood circulation and removing phlegm, and strengthening turbidity. It is composed of Salviae Miltiorrhizae Radix et Rhizoma 15 g, Persicae Semen 12 g, Angelicae Sinensis Radix 12 g, Achyranthis Bidentatae Radix 9 g, Rhei Radix et Rhizoma 15 g. Kangxianling can affect the synthesis and secretion of cytokines and inflammatory factors by expanding blood vessels, and can improve renal tubular fibrosis. It has a good multi-channel, multi-target and multi-directional protective effect on renal function. It also can delay the progress of chronic renal fibrosis by significantly alleviating such symptoms as fatigue and edema in patients with chronic renal fibrosis, and reducing serum creatinine, urea nitrogen and urine protein. In this paper, 5/6 nephrectomy, ischemia reperfusion injury, adriamycin-induced nephropathy, unilateral ureteral obstruction and other different modeling methods are listed. The mechanism of Kangxianling decoction in antagonizing renal fibrosis is discussed and summarized, which further provided new ideas and directions for future clinical and scientific research.

Optimization of preparation process of Jianpi Yishen Pill based on pharmacodynamic screening and orthogonal test / 中草药

Objective To screen the preparation technology of Jianpi Yishen Pill (JYP) by pharmacodynamics screening, and optimize it by orthogonal test to determine the best extraction technology. Methods The chronic renal failure model in rats was established by 5/6 nephrectomy. Three technological routes of JYP were selected, including A (powder of medicinal materials and decocting with water), B (powder of medicinal materials, extraction of volatile oil, and decocting with water), and C (extraction of volatile oil and decocting with water), with the content of Cr and BUN in rats as index. The orthogonal test was optimized by using the content of astragaloside and the quality of solid as indexes, with water volum, decoction time, decoction freguency as factors. Results The pharmacological efficacy test showed that technology A could obviously improve the rat model of chronic renal failure, and it could be used as the molding technology of this prescription pill. In this technology, five herbs such as Atractylodis Macrocephalae Rhizoma were crushed into fine powder alone, and the optimal extraction condition of other medical materials such as Astragalus Radix was as follows: 8-fold water per time, decocting 1 h, for two times. Conclusion JYP can obviously improve the content of Cr and BUN in rats model of 5/6 nephrectomy. The optimized preparation technology is stable and feasible, which can provide the basis for the follow-up development of JYP.

Effect of Shenfukang Ⅱ on kidney function and TGF-β1/Smads expression with 5/6nephrectomy in rats / 国际中医中药杂志

Objective To study the effect of Shenfukang Ⅱ on kidney function and TGF-β1/Smads signal pathway with 5/6 nephrectomy model in rats. Methods Thirty SD rats were randomly divided into the sham operation group, model group and Shenfukang group according to a random number table. We used 5/6 nephrectomy to prepare animal models. The rats were given Shenfukang Ⅱ particles 450 mg/(kg?d) gavage, the others group were given equal volume of saline gavage, once a day, continuous 21 days. The rats were given gavage at the 9 th week after operation. The changes of renal function index BUN, SCR and UA of rats in each group were detected 21 days after administration. The expression of TGF-β1, TGFβR1, Smad7, pSmad2, pSmad3 in renal tissues were detected by immunohistochemistry. The Smad7, pSmad2 and pSmad3 detected the expression of in renal tissues by Western Blot. Results Compared with the model group, the content of BUN (15.03 ± 2.19 mmol/L vs. 24.30 ± 2.82 mmol/L), SCR (74.22 ± 7.36 μmol/L vs. 93.61 ± 14.73 μmol/L) in Shenfukang group significantly decreased (P<0.05). The expression of TGF-β1, TGFβR1, pSmad2 and pSmad3 significantly decreased in kidney tissue, while the expression of Smad7 significantly increased in kidney tissue. The expression of p-smad2 (0.47 ± 0.03 vs. 0.64 ± 0.04) and p-smad3 (0.43 ± 0.04 vs. 0.55 ± 0.02) significantly decreased (P<0.05), and the expression of Smad7 (0.54 ± 0.03 vs. 0.39 ± 0.02) significantly increased (P<0.05). Conclusions Shenfukang Ⅱ have improved on kidney function with 5/6 nephrectomy model rats. The study revealed that Shenfukang II may play an anti-fibrotic effect on regulating TGF-β1/Smads signaling pathway with 5/6 nephrectomies model.

Protective Effects of Danshen Injection on 5/6 Nephrectomy Induced Model Rats with Inflammatory Renal Injury / 中国药房

OBJECTIVE:To study the protective effects of Danshen injection on 5/6 nephrectomiy induced model rats with in-flammatory renal injury. METHODS:50 rats were equally randomized into a sham-operation (isometric distilled water) group,a model (isometric distilled water) group,a positive control [captopril 5 mg/(kg·d)] group and Danshen injection low-dose and high-dose [1.6,4.8 ml/(kg·d)] groups. Models were established by performing 5/6 nephrectomy on the rats in all groups except the sham-operation group. The rats were given drugs for 4 consecutive weeks from the 6th week. 24 h urine protein was determined for all rats every week. After the last administration,the contents of creatinine,urea nitrogen and interleukins 1β(IL-1β),IL-2 and IL-10 in rats’serum were determined. Pathological changes and infiltration of macrophages CD68 in rats’kidney tissues were ob-served. RESULTS:Compared to the sham-operation group,those in the model group had more 24 h urine protein and more obvi-ous renal pathologic changes during 1-4 weeks of administration,and higher contents of creatinine,urea nitrogen,IL-1β,IL-2 and IL-10 and CD68 positive expression in serum. Compared to the model group,during 2-4 weeks of administration,those in the posi-tive control group and Danshen injection low-dose and high-dose groups had less 24 h urine protein,milder renal pathologic chang-es,and lower contents of creatinine,urea nitrogen,IL-1β and CD68 positive expression in serum,and higher content of IL-10, and only the rats in the positive control group demonstrated lower IL-2 content. There of the above were statistical differences(P<0.01 or P<0.05). CONCLUSIONS:Danshen injection can alleviate renal pathologic changes and reduce inflammatory injury of the kidneys of 5/6 nephrectomiy induced model rats.

Protective effect of telmisartan on rats with renal failure and its mechanism

Objective:To study the protective effect of telmisartan on rats with renal failure and its mechanism. Methods:60 Wistar rats were chosen as study objective, and were divided into 4 groups randomly:15 in group A (sham operation group), 15 in group B (model group), 15 in group C (telmisartan group) and 15 in group D (telmisartan+GW9962 group). The difference of survival rate, blood-urine biochemical indexes, renal pathological change, and the expression level of PPARγ and nNOS were compared. Results:After 12 weeks, the survival rate of group A was 93.33%(14/15), that of group B was 46.67%(7/15), that of group C was 86.67%(13/15), that of group D was 60.00%(9/15), and the difference among 4 groups had statistical significance (P0.05);after 3 weeks, 6 weeks and 12 weeks, these difference was statistical significant (P<0.05). The difference of blood-urine biochemical indexes, that of renal pathological change, and that of the expression level of PPAR毭and nNOS was statistical significant (P<0.05). Conclusions:Telmisartan has protective effect on renal failure caused by 5/6 nephrectomy, which might be relative to the expression level of PPARγ and nNOS.

Protective effect of telmisartan on rats with renal failure and its mechanism

Objective: To study the protective effect of telmisartan on rats with renal failure and its mechanism. Methods: 60 Wistar rats were chosen as study objective, and were divided into 4 groups randomly: 15 in group A (sham operation group), 15 in group B (model group), 15 in group C (telmisartan group) and 15 in group D (telmisartan+GW9962 group). The difference of survival rate, blood-urine biochemical indexes, renal pathological change, and the expression level of PPARγ and nNOS were compared. Results: After 12 weeks, the survival rate of group A was 93.33% (14/15), that of group B was 46.67% (7/15), that of group C was 86.67% (13/15), that of group D was 60.00% (9/15), and the difference among 4 groups had statistical significance (P0.05); after 3 weeks, 6 weeks and 12 weeks, these difference was statistical significant (P<0.05). The difference of blood-urine biochemical indexes, that of renal pathological change, and that of the expression level of PPARγ and nNOS was statistical significant (P<0.05). Conclusions: Telmisartan has protective effect on renal failure caused by 5/6 nephrectomy, which might be relative to the expression level of PPARγ and nNOS.

Protective effect of telmisartan on rats with renal failure and its mechanism

OBJECTIVE@#To study the protective effect of telmisartan on rats with renal failure and its mechanism.@*METHODS@#60 Wistar rats were chosen as study objective, and were divided into 4 groups randomly: 15 in group A (sham operation group), 15 in group B (model group), 15 in group C (telmisartan group) and 15 in group D (telmisartan + GW9962 group). The difference of survival rate, blood-urine biochemical indexes, renal pathological change, and the expression level of PPARγ and nNOS were compared.@*RESULTS@#After 12 weeks, the survival rate of group A was 93.33% (14/15), that of group B was 46.67% (7/15), that of group C was 86.67% (13/15), that of group D was 60.00% (9/15), and the difference among 4 groups had statistical significance (P  0.05); after 3 weeks, 6 weeks and 12 weeks, these difference was statistical significant (P < 0.05). The difference of blood-urine biochemical indexes, that of renal pathological change, and that of the expression level of PPARγ and nNOS was statistical significant (P < 0.05).@*CONCLUSIONS@#Telmisartan has protective effect on renal failure caused by 5/6 nephrectomy, which might be relative to the expression level of PPARγ and nNOS.

Comparison between the 5/6 nephrectomy SD rat and C57 mouse models of chronic renal failure / 中国比较医学杂志

Objective To find the similarities and differences between SD rat and C 57 mouse models of chronic renal failure established by 5/6 nephrectomy ,and select an appropriate experimental animal for the study of the treatment and prevention of chronic renal failure .Methods 60 SD rats and C57 mice were randomly and equally divided into 5/6 nephrectomy group and sham operatied group .The mice and rats were killed at 4 weeks , 8 weeks and 12 weeks after the operation, respectively.Serum and renal tissue samples were collected for the detection and analysis of renal function andα-SMA.Results There was no significant difference in the survival rates between the rat and mouse models (P>0.05). The serum creatinine and urea nitrogen in the 5/6NX rat group showed a stable state at 8-12 weeks, which was different from that of the 5/6NX mouse group.Conclusions A 5/6 nephrectomy mouse model of chronic renal failure is successfully established .Although both 5/6 NX mouse and rat groups appear progressive renal failure after the operation , their disease processes are not quite the same .In clinic, appropriate experimental animals should be selected according to the length of the observation period in order to accurately show the disease states .

The Anti-Inflammatory Effects of Statin in 5/6 Nephrectomized Rats / 대한내과학회지

BACKGROUND/AIMS: In a previous study, statin therapy reduced proteinuria and ameliorated the progression of chronic kidney disease. However, in patients with chronic renal failure (CRF), the beneficial effect of statin therapy on the preservation of renal function has not been determined. Thus, we determined the effect of rosuvastatin on CRF. METHODS: Male Sprague-Dawley rats (6 weeks old) were subjected to 5/6 nephrectomy. Six weeks after the procedure, the rats were divided into control and rosuvastatin-treated groups. Body weight and blood/urine biochemical parameters were measured 6 weeks after 5/6 nephrectomy and 8 weeks after the start of rosuvastatin treatment. Remnant kidneys were harvested at 6 (n = 4) and 14 (n = 8) weeks after 5/6 nephrectomy. RESULTS: During rosuvastatin treatment, changes in body weight and serum total and low-density lipoprotein cholesterol did not differ significantly between the control and rosuvastatin-treated groups. Although serum creatinine and proteinuria increased in both groups, the differences were not significant (p = 0.24 and 0.77, respectively). Immunohistochemical staining, enzyme-linked immunosorbent assays, and western blotting showed that the expression of transforming growth factor-beta1 and intracellular adhesion molecule-1 were reduced in the rosuvastatin-treated group. CONCLUSIONS: Long-term statin treatment may attenuate the inflammatory process in the progression of renal failure.

The Anti-Inflammatory Effects of Statin in 5/6 Nephrectomized Rats / 대한내과학회지

BACKGROUND/AIMS: In a previous study, statin therapy reduced proteinuria and ameliorated the progression of chronic kidney disease. However, in patients with chronic renal failure (CRF), the beneficial effect of statin therapy on the preservation of renal function has not been determined. Thus, we determined the effect of rosuvastatin on CRF. METHODS: Male Sprague-Dawley rats (6 weeks old) were subjected to 5/6 nephrectomy. Six weeks after the procedure, the rats were divided into control and rosuvastatin-treated groups. Body weight and blood/urine biochemical parameters were measured 6 weeks after 5/6 nephrectomy and 8 weeks after the start of rosuvastatin treatment. Remnant kidneys were harvested at 6 (n = 4) and 14 (n = 8) weeks after 5/6 nephrectomy. RESULTS: During rosuvastatin treatment, changes in body weight and serum total and low-density lipoprotein cholesterol did not differ significantly between the control and rosuvastatin-treated groups. Although serum creatinine and proteinuria increased in both groups, the differences were not significant (p = 0.24 and 0.77, respectively). Immunohistochemical staining, enzyme-linked immunosorbent assays, and western blotting showed that the expression of transforming growth factor-beta1 and intracellular adhesion molecule-1 were reduced in the rosuvastatin-treated group. CONCLUSIONS: Long-term statin treatment may attenuate the inflammatory process in the progression of renal failure.

Role of 11â-hydroxysteroid dehydrogenase 2 renal activity in potassium homeostasis in rats with chronic renal failure

Aldosterone concentrations vary in advanced chronic renal failure (CRF). The isozyme 11â-hydroxysteroid dehydrogenase 2 (11â-HSD2), which confers aldosterone specificity for mineralocorticoid receptors in distal tubules and collecting ducts, has been reported to be decreased or normal in patients with renal diseases. Our objective was to determine the role of aldosterone and 11â-HSD2 renal microsome activity, normalized for glomerular filtration rate (GFR), in maintaining K+ homeostasis in 5/6 nephrectomized rats. Male Wistar rats weighing 180-220 g at the beginning of the study were used. Rats with experimental CRF obtained by 5/6 nephrectomy (N = 9) and sham rats (N = 10) were maintained for 4 months. Systolic blood pressure and plasma creatinine (Pcr) concentration were measured at the end of the experiment. Sodium and potassium excretion and GFR were evaluated before and after spironolactone administration (10 mg·kg-1·day-1 for 7 days) and 11â-HSD2 activity on renal microsomes was determined. Systolic blood pressure (means ± SEM; Sham = 105 ± 8 and CRF = 149 ± 10 mmHg) and Pcr (Sham = 0.42 ± 0.03 and CRF = 2.53 ± 0.26 mg/dL) were higher (P < 0.05) while GFR (Sham = 1.46 ± 0.26 and CRF = 0.61 ± 0.06 mL/min) was lower (P < 0.05) in CRF, and plasma aldosterone (Pald) was the same in the two groups. Urinary sodium and potassium excretion was similar in the two groups under basal conditions but, after spironolactone treatment, only potassium excretion was decreased in CRF rats (sham = 0.95 ± 0.090 (before) vs 0.89 ± 0.09 µEq/min (after) and CRF = 1.05 ± 0.05 (before) vs 0.37 ± 0.07 µEq/min (after); P < 0.05). 11â-HSD2 activity on renal microsomes was lower in CRF rats (sham = 0.807 ± 0.09 and CRF = 0.217 ± 0.07 nmol·min-1·mg protein-1; P < 0.05), although when normalized for mL GFR it was similar in both groups. We conclude that K+ homeostasis is ...

Effects and Mechanism of Irbesartan on Tubulointerstitial fibrosis in 5/6 Nephrectomized Rats / 华中科技大学学报（医学）（英德文版）

Tubulointerstitial fibrosis(TIF)is a common pathological feature of end-stage kidney disease.Previous studies showed that upregulation of TGFβ1 notably contributed to the chronic renal injury and irbesartan halted the development of TIF in rats with 5/6 renal mass reduction.This study was to investigate the effects of irbesartan on chronic TIF and the mechanism involved TGFβ1 in the rodent model of chronic renal failure involving 5/6 nephrectomy.The results showed that irbesartan significantly attenuated the rise in blood pressure and tubulointerstitial injury observed in this model.Masson staining of the renal tissue revealed that there appeared severe renal tubule atrophy and fibrosis in operation group,but the lesion was attenuated mostly in irbesartan-treated group.Immunohistochemistry showed that irbesartan treatment apparently decreased the protein expression of TGFβ1 which was up-regulated in operation groups.Western blot showed that irbesartan treatment down-regulated the expression of TGFβ1,phosphorylated smad2(p-smad2),AT1R and phosphorylated p38(p-p38)MAPK,but significantly up-regulated the protein expression of smad6 as compared with operation group.These findings suggest that irbesartan attenuates hypertension and reduces the development of TIF in rats with 5/6 renal mass reduction via changes in the expression of these proteins at least including smad6,TGF-β1,p-smad2,AT1 and p-p38 MAPK.

The Role of Angiopoietin-1 and Thrombospondin-1 in the Kidney of Rats Subject to 5/6 Nephrectomy / 华中科技大学学报（医学）（英德文版）

The expression of angiopoietin-1 (Ang-1) and thrombospondin-1 (TSP-1) in 5/6 subtotal nephrectomy (STN) rats model, and its correlation to the renal microvasculature injury were investi-gated. Rat 5/6 STN model was established in adult male SD rats, and the sham-operated group and 5/6 STN group were set up. The renal function and histopathological changes were examined at the 1 st, 2nd, 4th, 8th and 12th week after operation. The expression of Ang-1, TSP-1 and CD31 in renal tissues was detected by using immunohistochemistry. From 2nd to 8th week after operation, Ang-1 was significantly expressed in glomeruli of rats with STN. Ang-1 staining in glomeruli of STN group was increased significantly as compared with that in sham-operated group at 4th and 8th week after operation, and subsequently decreased after the 12th week. The expression of TSP-1 was increased significantly in STN group. As compared with sham-operated group, the CD31 expression was sig-nificantly down-regulated from the 2nd week. The expression orAng-1 mRNA was detected by using RT-PCR at the same time points. The expression of Ang-1 mRNA in renal tissue of rats with STN was significantly up-regulated at the 2rid, 4th and 8th week after operation as compared with that in STN group at other time points or in sham-operated group at the same time points, while decreased evidently at the 12th week as compared with that in sham-operated group. It is concluded that there are changes in the mRNA expression of Ang-1, and the significant up-regulation of the expression of TSP-1 in renal tissue of rats with STN, which may be involved in the remnant renal microvasculature injury.

Apoptosis Signaling Pathway in a Subtotal Nephrectomy Rat Model / 华中科技大学学报（医学）（英德文版）

To investigate the role and mechanisms of apoptosis and apoptosis signaling pathway in 5/6 nephrectomy rat model (SNx), the mRNA and protein levels of caspase-3, -8, -9 and apoptosis were detected by in situ end labeling ( TUNEL ), immunohistochemistry, RT-PCR, Western-blotting 1,2, 4, 8, 12, 16, 26 and 40 weeks after 5/6 nephrectomy rat model was made respectively. The rats in the model group developed glomerular sclerosis and renal interstitial fibrosis. The number of the apoptototic cells in glomeruli, renal tubule and renal interstitium was remarkably higher in the model group than that in the control group (P＜0.05, P＜0.01). Changes of mRNA and protein level of caspase-3, -8, -9 had the same tendency and was up-regulated wavily in the rat model compared with the control group (P＜0.05). Peaks in model appeared on the 4th and the 40th week respectively. The growth amplitude of caspase-9 was remarkably higher than that of caspase-8. It is concluded that the development of 5/6 nephrectomy rat model was correlated with the apoptosis of glomeruli, renal tubule and renal interstitium. Both of death receptor and mitochondria signaling pathways are involved in the process and the latter might play a primary role.

The Role of Renin-Angiotensin System in Progressive Renal Injury / 대한신장학회잡지

Angiotensin II(A II) -a main effector molecule of renin-angiotensin system(RAS) has been known to increase blood pressure and glomerular capillary pressure, and filtration fraction which may be involved in the progressive renal injury process. The action of A II takes place mainly through AT1 receptor. RAS can be blocked by angiotensin converting enzyme inhibitor(ACEI) and recently developed A II AT1 receptor antagonist(AT1 RA). ACEI also activate kinin system, simultaneously. However, AT1 RA does not affect kinin system. The renoprotective mechanism of ACEI may be related with activation of kinin system. In order to evaluate the renoprotective mechanism of long-term ACEI(enalapril, 100mg/L in drinking water for 12 weeks) or AT1 RA treatment(losartan 300mg/L in drinking water for 12 weeks), and its effect on the cytokines and growth factor expressions of renal cortical tissue by compatitive RT-PCR, 46 5/6 nephrectomized(5/6 NPX) rats and 8 sham operated rats were included in this study. Five sixth NPX rats showed marked hypertensin, significant proteinuria and glomerulosclerosis(mean 30.5%) in 12 weeks after surgery. However, enelapril or losartan treated rats revealed significantly lower 24 hour urinary protein excretion(UProtV), systolic blood pressure(SBP), and glomerulosclerosis than those of control 5/6 NPX rats. Plasma renin activity and angiotensin II levels of 5/6 NPX untreated control rats were not significantly increased compared to sham operated rats in 12 week after surgery. Renal cortical renin gene expression of untreated 5/6 NPX rats was significantly suppressed compared to sham operated rats. Enalapril or losartan treated 5/6 NPX rats showed significantly increased renin gene expression compared to untreated 5/6 NPX rats. Renal cortical gene expressions of TGF-beta, TNF-alpha, MCP-1, IL-6, osteopontin, and endothelin-1 were significantly increased in 5/6 NPX untreated control rats compared to sham operated rats. Enalapril or losartan treated 5/6 NPX rats showed significantly less level of renal TGF-beta gene expression compared to 5/6 NPX control rats. The magnitude of SBP and UProtV were significantly positively correlated with the degree of glomeruloslerosis(p<0.001, p<0.001). With the above result, we speculate that because ACEI or AT1 RA showed similar renoprotective effect in 5/6 NPX rats, at least in part, local activation of RAS plays an important role in the progressive renal injury process of this model.

The Role of Renin-Angiotensin System in Progressive Renal Injury / 대한신장학회잡지

Angiotensin II(A II) -a main effector molecule of renin-angiotensin system(RAS) has been known to increase blood pressure and glomerular capillary pressure, and filtration fraction which may be involved in the progressive renal injury process. The action of A II takes place mainly through AT1 receptor. RAS can be blocked by angiotensin converting enzyme inhibitor(ACEI) and recently developed A II AT1 receptor antagonist(AT1 RA). ACEI also activate kinin system, simultaneously. However, AT1 RA does not affect kinin system. The renoprotective mechanism of ACEI may be related with activation of kinin system. In order to evaluate the renoprotective mechanism of long-term ACEI(enalapril, 100mg/L in drinking water for 12 weeks) or AT1 RA treatment(losartan 300mg/L in drinking water for 12 weeks), and its effect on the cytokines and growth factor expressions of renal cortical tissue by compatitive RT-PCR, 46 5/6 nephrectomized(5/6 NPX) rats and 8 sham operated rats were included in this study. Five sixth NPX rats showed marked hypertensin, significant proteinuria and glomerulosclerosis(mean 30.5%) in 12 weeks after surgery. However, enelapril or losartan treated rats revealed significantly lower 24 hour urinary protein excretion(UProtV), systolic blood pressure(SBP), and glomerulosclerosis than those of control 5/6 NPX rats. Plasma renin activity and angiotensin II levels of 5/6 NPX untreated control rats were not significantly increased compared to sham operated rats in 12 week after surgery. Renal cortical renin gene expression of untreated 5/6 NPX rats was significantly suppressed compared to sham operated rats. Enalapril or losartan treated 5/6 NPX rats showed significantly increased renin gene expression compared to untreated 5/6 NPX rats. Renal cortical gene expressions of TGF-beta, TNF-alpha, MCP-1, IL-6, osteopontin, and endothelin-1 were significantly increased in 5/6 NPX untreated control rats compared to sham operated rats. Enalapril or losartan treated 5/6 NPX rats showed significantly less level of renal TGF-beta gene expression compared to 5/6 NPX control rats. The magnitude of SBP and UProtV were significantly positively correlated with the degree of glomeruloslerosis(p<0.001, p<0.001). With the above result, we speculate that because ACEI or AT1 RA showed similar renoprotective effect in 5/6 NPX rats, at least in part, local activation of RAS plays an important role in the progressive renal injury process of this model.