Effects of Compound Shenhua Tablet (复方肾华片) on Renal Tubular Na+-K+-ATPase in Rats with Acute Ischemic Reperfusion Injury

Objective: To investigate the protective effects and potential mechanisms of Shenhua Tablet(肾华片, SHT) on the toll-like receptors(TLRs)-mediated signaling pathways in a rat model of kidney ischemia-reperfusion injury(IRI). Methods: Sixty male Wistar rats were randomly divided into 5 groups: sham surgery, model control, astragaloside(150 mg·kg~(-1)·d~(-1)), low-and high-dose SHT(1.5 and 3.0 g·kg~(-1)·d~(-1), repectively) groups. One week after drug treatment, rats underwent surgery to establish the IRI models. At 24 h and 72 h after the modeling, serum creatinine(Scr) and blood urea nitrogen(BUN) were analyzed; pathological damage were scored after periodic acid-Schiff staining. TLR2, TLR4 and myeloid differentiation factor 88(My D88) protein and m RNA expressions were detected by inmmunohistochemistry, Western blot and qPCR. Tumor necrosis factor α(TNF-α) and interleukin-6(IL-6) protein expressions were detected by enzyme linked immunosorbent assay. Results: Compared with the sham group, the model group exhibited severe change in renal function(Scr: 189.42±21.50, P0.05), pathological damage(damage score: 4.50±0.55, P0.05), and the expression levels of TLR2, TLR4, MyD88, TNF-α, IL-6 were significantly higher than other groups. Meanwhile, the levels of TLRs in model group showed upward tendency from 24 to 72 h, unparalleled with pathological and functional changes. The aforementioned parameters were alleviated to a certain extent, and, in addition to TLRs, presented the obvious downward trending from the 24 to 72 h after the intervention in the SHT and astragaloside groups relative to the model(P0.05); in particular, the most significant mitigation of these changes was observed in the SHT-H group(P0.05). Conclusions: TLRs may be an important spot to treat and research in acute kidney injury. SHT could effectively mitigate renal injuries and promote recovery of IRI injuries through suppression of degeneration induced by up-regulation of TLR2 and TLR4 expression levels in the My D88-dependent signaling pathway and exhibit some dose dependence.     

Effect of Xuefu Zhuyu Capsule (血府逐瘀胶囊) on Angiogenesis in Hindlimb Ischemic Rats

Objective: To investigate the effect and mechanism of Xuefu Zhuyu Capsule(血府逐瘀胶囊, XZC) on pro-angiogenesis in the hindlimb ischemic model rats. Methods: A total of 100 Sprague Dawley rats were randomly divided into a model group, a regular-dose XZC group(0.48 g·kg~(-1)·d~(-1)) and a high-dose XZC group(0.96 g·kg~(-1)·d~(-1)) using random number table method. The model of hindlimb ischemic rats were made through femoral artery embolization with Bletilla microsphere agent. XZC were given on the first day after embolization surgery and lasted 5 days. Finally 72 models were obtained with 12 in each group for each time point. The lower ischemic limb was amputated on the third day after embolization surgery. Histopathological characters and the number of blood vessels of granulation tissues were observed at 36 and 48 h after amputation, respectively. The main genes were obtained from microarray analysis and were validated using real-time quantitative polymerase chain reaction. Results: The vascular number of granulation tissues at both 36 and 48 h were characterized by new and fresh vessels. The number of angiogenesis in the high-dose XZC group at 36 and 48 h was greater compared with that in the regular-dose XZC and model groups(P0.01), and high-dose XZC at 36 h increased more vessels than that at 48 h(P0.01). Consequently, granulation tissues from the high-dose XZC group at 36 h were chosen for microarray analysis. In all, 2,085 differentially expressed genes(DEGs) were detected and 25 DEGs were determined to be directly related to angiogenesis. Four biological process terms were found including angiogenesis, regulation of angiogenesis, positive regulation of angiogenesis, and positive regulation of vascular endothelial growth factor receptor signaling pathway(P0.05). Microarray analysis also showed 49 pathways including 11 pathways related to angiogenesis. Conclusion: XZC promoted angiogenesis moderately and the mechanism involved multiple DEGs and multiple pathways.     

Extracellular ascorbic acid fluctuation during the protective process of ischemic preconditioning in rabbit renal ischemia-reperfusion model measured

Background Ascorbic acid has important antioxidant ischemic preconditioning on later ischemia-reperfusion ascorbic acid in ischemic preconditioning in the kidney. properties, and may play a role in the protective effects of Herein, we examined the role of endogenous extracellular Methods We developed a solitary rabbit kidney model where animals received ischemia-reperfusion only （ischemia-reperfusion group, n=-15） or ischemic preconditioning followed by ischemia-reperfusion （ischemic preconditioning group, n=15）. Ischemia-reperfusion was induced by occluding and loosening of the renal pedicle. The process of ischemic preconditioning included 15-minute brief ischemia and 10-minute reperfusion. In vivo microdialysis coupled with online electrochemical detection was used to determine levels of endogenous extracellular ascorbic acid in both groups. The extent of tissue damage was determined in kidney sections stained with hematoxylin and eosin. Serum creatinine and urea nitrogen were also detected to assess renal function. Results During ischemia-reperfusion, the extracellular ascorbic acid concentration during ischemia increased rapidly to the peak level （（130.01±9.98）%）, and then decreased slowly to near basal levels. Similar changes were observed during reperfusion （peak level, （126.78±18.24）%）. In the ischemic preconditioning group there was a similar pattern of extracellular ascorbic acid concentration during ischemic preconditioning. However, the ascorbic acid level was significantly lower during the ischemia and early reperfusion stage compared to the ischemia-reperfusion group. Additionally, the extent of glomerular ischemic collapse, tubular dilation, tubular denudation, and loss of brush border were markedly attenuated in the ischemic preconditioning group. Levels of serum creatinine and urea nitrogen were also decreased significantly in the ischemic preconditioning group. Conclusions Ischemic preconditioning may protect renal tissue against ischemia-reperfusion injury via use of extracellular ascorbic acid. In vivo microdialysis coupled with online electrochemical detection is effective for continuous monitoring extracellular ascorbic acid in the renal cortex.     

Structural Modulation of Gut Microbiota in Rats with Allergic Bronchial Asthma Treated with Recuperating Lung Decoction

ObjectiveTo investigate whether recuperating lung decoction (RLD) can modulate the composition of gut microbiota in rats during asthma treatment. MethodsFifteen Sprague-Dawley rats were divided randomly and equally into control group, model group, dexamethasone (DEX) group, RLD medium-dose group, and RLD high-dose group. The asthma model was established in all groups, except for the control group. The rats in the DEX and RLD groups were treated orally with DEX and RLD, respectively. The rats in the control and model groups were treated orally with 0.9% saline. The intestinal bacterial communities were compared among groups using 16S rRNA gene amplification and 454 pyrosequencing. ResultsThe microbial flora differed between the control and model groups, but the flora in the RLD groups was similar to that in the control group. No significant differences were observed between the RLD high-dose and medium-dose groups. RLD treatment resulted in an increase in the level beneficial bacteria in the gut, such asLactobacillusandBifidobacteriumspp. ConclusionOral administration of RLD increased the number of intestinal lactic acid-producing bacteria, such as Lactobacillus andBifidobacterium, in asthma model rats.     

Therapeutic effect of bFGF on retina ischemia-reperfusion injury

Background Basic fibroblast growth factor (bFGF) plays important roles in retina degeneration, light injury, mechanical injury, especially in retina ischemia-reperfusion injury (RIRI). This study was to investigate the therapeutical effect of bFGF on RIRI and its mechanisms.Methods Experimental RIRI was induced by increasing intraocular pressure (IOP) in the eyes of 48 rats. These rats were divided into normal control, ischemia-reperfusion and bFGF-treated groups.Histological and ultrastructural changes of in the retina of different groups were observed, and the number of retinal ganglion cells (RGCs) was quantitatively analyzed under microscopy. Apoptotic cells were detected using the TdT-dUTP terminal nick-end labeling (TUNEL) method. The expression of caspase-3 was determined by streptavidin peroxidase (SP) immunohistochemistry. Atomic absorption spectrum method was used to evaluate the intracellular calcium changes.Results At the early stage of retinal ischemia-reperfusion injury, retina edema in the treated group was significantly eliminated compared with the untreated ischemic animals. RGCs in the bFGF-treated group was more than those in the untreated ischemic group during the post-reperfusion stages. In ischemic group, apoptotic cells could be found at 6th hours after reperfusion and reached the peak at 24th hours. At 72th hours no apoptotic cells could be found. The changes in caspase-3 expression had a similar manner. The intracellular calcium of rat retina began to increase at lth hour, reached the peak at 24 hours, and began to decease at 72th hours. The change of the three markers in the treatment group showed a similar pattern, but they were all relatively less obvious.Conclusion Apoptosis may play a vital role in RIRI. bFGF may has therapeutical effects on RIRI by inhibiting the increase of intracellular calciums and caspase-3 expression.     

Effects of Fufang Shenhua Tablet(复方肾华片) on the Expression of Toll-Like Receptors during Acute Kidney Injury Induced by Ischemia-Reperfusion in Rats

<正>Objective:To investigate the impact of a traditional Chinese medicinal compound known as Fufang Shenhua Tablet(复方肾华片,SHP) on the expression of Toll-like receptors(TLRs) during renal ischemia-reperfusion injury(IRI)-induced acute kidney injury(AKI) in rats.Methods:A total of 28 Wistar rats were randomly divided into five groups:(1) pseudo-operation control group,(2) ischemia-reperfusion model group, (3) Astragaloside group,(4) high-dose SHP group,and(5) low-dose SHP group.There were four rats in the pseudo-operation group and six rats in each of the other groups.The accepted ischemia-reperfusion model was established after a 7-day gavage intervention,and pathological changes and renal function were observed,using an enzyme-linked immunosorbent assay(ELISA) to detect interleukin 8(IL-8) and interferon gamma(IFN-γ) levels,as well as immunohistochemical staining to detect altered levels of TLR2 and TLR4 expression in renal tissue.Results:After 24 h,renal pathological damage and the expression levels of serum creatinine(Scr),IL-8, IFN-γ,TLR2,and TLR4 were significantly higher in the model group as compared with the pseudo-operation group(P0.05).In addition,at 24 h the above indicators decreased significantly in the Astragaloside group,high-dose SHP group and low-dose SHP group as compared with the ischemia-reperfusion model group(P0.05). TLR2 and TLR4 expression levels were significantly reduced in the SHP treatment and Astragaloside group as compared with the pseudo-operation group(P0.05).Further,the high-dose SHP group showed significantly less renal damage score and decreased levels of TLR expression than those of low-dose SHP group and Astragaloside group(all P0.05).Conclusion:SHP can alleviate the renal structural and functional damage caused by IRI-induced AKI in rats by reducing the damage of renal pathology,which may reduce inflammatory cytokine levels by downregulating the expression of TLRs in renal tissue in a dose-dependent manner.     

The role of angiopoietin-1 and thrombospondin-1 in the kidney of rats subject to 5/6 nephrectomy

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 investigated. 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 1st, 2nd, 4th, 8th and 12th week after operation. The expression orAng-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 significantly down-regulated from the 2nd week. The expression of Ang-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 2nd, 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.     

EFFECTS OF ISCHEMIA ON CEREBRAL Na~+, K~+-ATPase ACTIVITY IN THE RABBITS SUBJECTED TO MCAo

Changes in the activity of Na~+, K~+-ATPase and in the water, sodium, and potassium levels in the ischemic brain were investigated in rabbits 2, 4, and 24h following occlusion of middle cerebral artery (MCAo) and in shamoccluded control. An increase in Na~+, K~+-ATPase activity was observed in 2h and 4h groups, with a subsequent decrease in the enzyme activity. The elevation in Na~+, K~+-ATPase activity was accompanied by an increase in the sodium content and a slight decrease in the potassium content. These changes are presumed to occure because of stimilated active transport of sodium from blood to brain across the brain capillaries. We suggest that the elevated activity of Na~+, K~+-A TPase may participate in the pathogenesis of ischemic brain edema.     

Significance of Rosiglitazone Inhibiting TLR4 Expression in Partial Hepatic Ischemia/Reperfusion of Mice

The effect of rosiglitazone as the ligand of peroxisome proliferator-activated receptor γ (PPARγ) inhibiting the TLR4 expression in ischemic lobes in partial hepatic ischemia/reperfusion injury (IRI) in BABL/C mice and the action of rosiglitazone inhibiting the TLR4 receptor-mediated inherent immune response were investigated. The model of the mouse partial hepatic ische- mia/reperfusion injury was established. All the animals were randomly divided into 3 groups: rosiglitazone group, vehicle (dimethylsulphoxide, DMSO) group and sham operation group. The hepatic samples were collected when mice were sacrificed 0, 2, 4 and 6 h after reperfusion following 1 h ischemia to analyze the acute phase of hepatic IRI. The dynamic expression of TlR4 mRNA was de- tected quantitatively by real-time-PCR, and the levels of TNF-α, IL-10 and ALT in portal vein were determined in all groups. After restoration of blood supply, the expression of TLR4 mRNA in ischemic lobes was detected in 0, 2, 4 and 6 h after reperfusion following 1 h ischemia in rosiglitazone group and vehicle group. The most intensive expression of TLR4 mRNA was present at 4 h after reperfusion in ischemic lobes in vehicle group. As compared with vehicle group, the expression of TLR4 mRNA in ischemic lobes in rosiglitazone group was significantly decreased at 4 h after reper- fusion. The level of IL-10 in portal vein was markedly up-regulated in rosiglitazone group as compared with vehicle group. Contrarily, the levels of TNF-α and ALT in portal vein were markedly down-regulated in rosiglitazone group as compared with vehicle group at every time point in mouse partial hepatic IRI model. Rosiglitazone could alleviate the hepatic IRI by inhibiting TLR4 receptor-mediated inherent immune response.     

Effect of Quyu Chencuo Formula(去菀陈莝方)on Renal Fibrosis in Obstructive Nephropathy Rats

Objective: To observe the effect of Quyu Chencuo Formula(去菀陈莝方, QCF) on renal fibrosis in rats with obstructive nephropathy. Methods: Twenty-four rats were randomly divided into three groups, 4 for sham operation as the control group, 10 for unilateral ureteral obstruction(UUO) model group, and the rest 10 for QCF treating UUO model group. All rats were sacrificed under 3% pentobarbital(50 mg/kg) anesthesia on the 14 th day after surgery, then the right kidney samples of rats were harvested for hematoxylin eosin(HE) staining and Masson staining to observe the renal pathological changes. Immunohistochemistry and Western blotting were used to examine the expression of transforming growth factor β1(TGF-β1), and real-time polymerase chain reaction(RT-PCR) was employed to examine the expressions of TGF-β1, α-smooth muscle actin(α-SMA) and E-cadherin mRNA. Results: HE and Masson staining showed that the renal interstitial of the rats in the control group had no significant fibrotic lesion; in the model group, there were obvious interstitial fibrosis; for the QCF group, there were epithelial cell necrosis, infiltration of lymphocytes and mononuclear cells, aggravated interstitial fibrosis in varied degrees, but the pathological changes were less in the QCF group than in the model group. The immunohistochemistry and Western blotting results showed that the TGF-β1 expression was increased significantly in the model group, while decreased significantly in the QCF group(P0.05); RT-PCR showed that the mRNA expression of α-SMA and TGF-β1 increased significantly in the model group, while both were significantly decreased in the QCF group compared with the model group(P0.05). The mRNA expression of E-cadherin was decreased significantly in the model group, and it was significantly increased in the QCF group as compared with the model group(P0.05). Conclusion: QCF may improve renal fibrosis by regulating the expressions of TGF-β1, α-SMA and E-cadherin, and prevent the progress of kidney fibrosis.     

Effect of Yishen Huayu Fang on kidney tissue E-cadherin expression in unilateral ureter ligation in rats

OBJECTIVE:To observe E-calcium sticky protein(E-cadherin) expression in kidney tissues in a rat model of unilateral ureter ligation and the effect of Yishen Huayu Fang(formula of tonifying the kidney and dissolving accumulated blood stasis) on the expression.METHODS:A total of 150 clean grade male rats were randomly divided into a control group,model group,low-dose Yishen Huayu Fang group(low-dose group),high-dose Yishen Huayu Fang group(high-dose group),and Lotensin group.A renal fibrosis model was established with unilateral ureteral obstruction(UUO).Pathological changes of rat renal tissue were observed with light microscopy on days 3,7,14,21,and 28 after UUO.Changes in kidney tissue E-cadherin expression were observed with immunohistochemistry.RESULTS:Three days after modeling,kidney edema appeared followed by gradual inflammatory cell infiltration,and part of the small tubules disappeared while the renal cortex thinned.Meanwhile,the E-cadherin expression level dropped,which was negatively correlated with the obstruction time.After intervention,E-cadherin expression was increased in all treatment groups(P<0.01 or P<0.05),while there were no significant differences between the high-dose and Lotensin groups.CONCLUSION:Yishen Huayu Fang delays the renal fibrosis process by promoting E-cadherin expression in renal tissues and reducing extracellular matrix deposition.     

EFFECTS OF PHYCOCYANIN ON EXPRESSION OF CytC mRNA AND CASPASE-3 mRNA AFTER FOCAL CEREBRAL ISCHEMIA／REPERFUSION IN RATS

Objective To study the effects of phycocyanin on the expression of Cytochrome C (CytC) genes and Caspase-3 genes after focal cerebral ischemia/reperfusion in rats. Methods A rat middle cerebral artery occlusion (MCAO)/reperfusion model was produced using the intraluminal filament method. The rats were divided into three groups: sham operation group, model control group and phycocyanin group. After MCAO, the neurobehavioral testing of all rats was made. The infarction area was evaluated with the method of 2,3,7-triphenylt-etrazolium chloride (TTC) staining. The expression of CytC mRNA and Caspase-3 mRNA were determined by in situ hybridization. Results In the sham operation group and the model control group, there was only a few CytCpositive cells were seen in the normal cerebral tissue. In the model control group, the upregulation of CytC mRNA began 6h after ischemia, reached a maximum at 12h (cortex)-24h (striatum) , then subsided gradually, but still in high level. In the phycocyanin group, CytC-positive cells were also mainly in cortex and striatum, but the number of the cells was significantly lower than the number of the model control group. The time-phase pattern of CytC mRNA in the phycocyanin group was similar to the pattern of the model control group. In the sham operation group and the model control group, there was only a few Caspase-3-positive cells were seen in the normal cerebral tissue. In the model control group, the upregulation of Caspase-3 mRNA began 6h after ischemia, reached a maximum at 24h and subsided at 48h, but still in high level. In the phycocyanin group, Caspase-3-positive cells were also mainly in the penumbral area, but the number of the cells were significantly lower than the number of the model control group. The time-phase pattern of Caspase-3 mRNA in the phycocyanin group was similar to the pattern of the model control group. Conclusion The over-expression of CytC mRNA and Caspase-3 mRNA might play a key role in ischemic cerebral injury after MCAO. Phycocyanin could inhibit the over-expression of CytC mRNA and Caspase-3 mRNA in the cerebral cortex, and might play an important role in the protection of ischemic neurons.     

Kyoto Breast Cancer Consensus Conference

Background To date, there have been no reports on altered nitric oxide （NO） content in ischemia/reperfusion with regard to in vivo preconditioning procedures. These studies are important for understanding the mechanisms of NO during early myocardial ischemic preconditioning. The aim of the present study was to investigate the mechanisms of NO during early myocardial ischemic preconditioning by measuring levels of NO and cyclic guanosine monophosphate （cGMP）, as well as activity of nitric oxide synthase （NOS） in ischemia/reperfusion with respect to preconditioning in rats.
Methods Sixty-six female Sprague-Dawley rats were randomly divided into four groups： ischemic preconditioning group （IP）, ischemia/reperfusion group （I/R）, control group （CON）, and preconditioning procedure group （PC）. In the PC group, rats were further divided into PC1-, PC1＋, PC2-, PC2＋, PC3-, and PC3＋ subgroups. Rats underwent left coronary artery occlusion and reperfusion, and subsequently, NOS activity and levels were assessed with spectrophotometric analysis. cGMP contents were measured with radioimmunoassay.
Results The level of NO and cGMP, as well as the activity of NOS, were significantly higher in the IP group compared to the I/R and CON groups （P 〈0.05）. During preconditioning prior to prolonged ischemia, NO and cGMP levels varied markedly with ischemia and reperfusion. The levels of NO repeatedly increased when the heart was exposed to three episodes of 5-minute ischemia, and were almost completely reversed during each reperfusion period. NO and cGMP levels were significantly different between the 5-minute period of ischemia and the same period of reperfusion during preconditioning.
Conclusions NO plays an important role during early phase myocardial ischemic preconditioning in rats. NO and cGMP could be triggers and mediators of early phase myocardial ischemic preconditioning. Altered NOS activity following ischemic stress could be the primary inducer of higher NO levels detected. NO     

Dynamic expression of glucose transporters 1 and 3 in the brain of diabetic rats with cerebral ischemia reperfusion

Background Blood glucose control improves the outcome of diabetic patients with stroke, but the target range of blood glucose control remains controversial. The functional recruitment of ischemia penumbra is extremely important to the recovery after stroke. The present study aimed to explore the expression of brain-type glucose transporters （GLUT1 and GLUT3） in cerebral ischemic penumbra at different blood glucose levels and different ischemic-reperfusion time in diabetic hypoxia-ischemia rats. The results might provide an experimental basis for clinical treatment of diabetic patients with stroke. Methods The Wistar rats included in this study were randomly assigned to 4 groups （50 rats each）： normal control group （NC）, uncontrolled diabetic group （DM1）, poorly-controlled diabetic group （DM2）, and well-controlled diabetic group （DM3）. Diabetic rats were induced by single intraperitoneal injection of streptozotocin, and the focal ischemic rat model of middle artery occlusion （MCAO） was made by insertion of fishing thread in 6 weeks after the establishment of the diabetic model. Each group was divided into 5 subgroups （10 rats each）： four focal ischemic subgroups at different ischemic-reperfusion time （at 3,12, 24 and 72 hours after reperfusion, respectively） and one sham-operated subgroup. The mRNA and protein expression of GLUT1 and GLUT3 was assessed by RT-PCR and Western blotting, respectively. Results There was significant difference in the mRNA expression of GLUT1 and GLUT3 between the four focal ischemic subgroups and the sham-operated subgroup at different reperfusion time in each group. The mRNA expression of GLUT1 and GLUT3 in the 4 ischemic groups began to increase at 3 hours, peaked at 24 hours after reperfusion and maintained at a higher level even at 72 hours compared with that of the sham-operated subgroup. The mRNA expression of GLUT1 increased more significantly than that of GLUT3. The mRNA expression of GLUT1 and GLUT3 was significantly different between the diabetic groups and normal control group. The mRNA expression of GLUT1 and GLUT3 was increased more significantty in the diabetic groups than that in the normal control group. There was a significant difference in the mRNA expression in the groups with different blood glucose levels. The mRNA expression tended to decrease with increased blood glucose levels. The expression trend of GLUT1 and GLUT3 protein was similar to that of GLUT1 and GLUT3 mRNA. Conclusions GLUT1 and GLUT3 expression was notably up-regulated in the penumbra region after cerebral ischemia in this study. But the up-regulated amplitude of GLUT1 and GLUT3 in the diabetic rats with cerebral ischemic injury became smaller than that of the normal controls. In the treatment of diabetic patients with cerebral embolism, blood glucose control should not be too strict, otherwise the up-regulation of GLUT1 and GLUT3 induced by cerebral ischemic injury might not be able to meet the needs of energy metabolism in cells. Chin Med J 2009; 122（ 17）： 1996-2001     

A Comparative Study on Several Models of Experimental Renal Calcium Oxalate Stones Formation in Rats

In order to compare the effects of several experimental renal calcium oxalate stones formation models in rats and to find a simple and convenient model with significant effect of calcium oxalate crystals deposition in the kidney, several rat models of renal calcium oxalate stones formation were induced by some crystal-inducing drugs (CID) including ethylene glycol (EG), ammonium chloride (AC), vitamin D3 [1α(OH)VitD3, alfacalcidol], calcium gluconate, ammonium oxalate, gen-tamicin sulfate, L-hydroxyproline. The rats were fed with drugs given singly or unitedly. At the end of experiment, 24-h urines were collected and the serum creatinine (Cr), blood urea nitrogen (BUN), the extents of calcium oxalate crystal deposition in the renal tissue, urinary calcium and oxalate ex-cretion were measured. The serum Cr levels in the stone-forming groups were significantly higher than those in the control group except for the group EG L-hydroxyproline, group calcium gluconate and group oxalate. Blood BUN concentration was significantly higher in rats fed with CID than that in control group except for group EG L-hydroxyproline and group ammonium oxalate plus calcium gluconate. In the group of rats administered with EG plus Vitamin D3, the deposition of calcium ox-alate crystal in the renal tissue and urinary calcium excretion were significantly greater than other model groups. The effect of the model induced by EG plus AC was similar to that in the group in-duced by EG plus Vitamin D3. EG plus Vitamin D3 or EG plus AC could stably and significantly in-duced the rat model of renal calcium oxalate stones formation.     

Astragali and Angelica protect the kidney against ischemia and reperfus ion injury and accelerate recovery

Objective To investigate the role of Astragali and Angelica (A&amp;A) of Chinese herbs in acut e renal ischemia/reperfusion injury, and the related intracellular signal transd uction mechanism.Methods Acute ischemic renal injury in rats was induced by clamping in renal pedicel for 45 minutes. Rats in therapy group were given a single dose （2 ml/day） of A&amp; A for 3 days before clamping, and then continued for another 3 days. Forty-fiv e minutes after clamping and at different reperfusion time, serum creatinine (Scr) and renal pathological changes were taken and compared in both groups. The p roliferating cell nuclear antigen (PCNA) positive cells were detected by immunoh istochemistry. Extracelluar regulating kinase (ERK) and c-Jun N-terminal kina se (JNK) activity was assayed by specific substrate phosphorylation with immunop recipitation. Results At the 24th hour of reperfusion, Scr was lower in A&amp;A group than that in the con trol. Much less necrotic tubular cells, casts, and more PCNA-positive cells were found in A&amp;A group. ERK activity decreased after clamping, and recovered a t 5 minutes of reperfusion. There was no difference between the two groups. JN K activity did not change after ischemia, but increased at 5 minutes and peaked at 20 minutes of reperfusion. JNK activity was significantly higher in A&amp;A group than that in the control group. Conclusion A&amp;A protected kidney against ischemic insult and accelerated both functional and histological recovery after acute renal ischemia/reperfusion injury, which may associate with the change of JNK signaling pathway.     

Bone Marrow-Derived Mesenchymal Stem Cells Incubated with Tanshinone IIA Aattenuates Learning and Memory Impairment Induced by β-amyloid and its Underlying Mechanisms in Rats

Objective：The present study was designed to investigate the neuroprotective effect of mesenchymal stem cells （MSC） which incubated with tanshinone IIA on β-amyloid （Aβ）-induced learning and memory impairment in rats,and further explore the underlying potential mechanisms. Methods：60 healthy male SD rats were randomly divided into 4 groups：sham-operated group,model group,MSC group,MSC+tanshinone group IIA（ n=15）.The rats of model group were injected into the hippocampal with aggregation of 5 μL Aβ25-35（ 2 μg·μL-1） to establish the model of learning and memory impairment. 1 week after surgery,the MSC were injected into the hippocampus,while the rats of model group were injected with volume-matched DMEM,instead. 2 weeks after surgery, Morris water maze （MWM） test was applied to evaluate spatial learning and memory ability of rats,then the rats were sacrifi ced. The survival status of hippocampal neurons was detected by HE and Tunel staining;the formation of hippocampal senile plaques was detected by thiofl avin S staining;the protein expression of Aβ1-42,p-Tau and AMPK in mouse hippocampus were detected by Western blot,respectively;the expression of AMPK mRNA was detected by PCR and the activity of SOD,T-AOC,GSH-PX and content of MDA in plasma by the kits.Results：Compared with sham-operated group, the mean escape latency was markedly increased and the time percentage in target quadrant showed notable decrease in model group;large number of the formation of senile plaque in the hippocampus was detected;the number of surviving neurons in hippocampus was signifi cantly decreased and the apoptosis increased;the protein expression of Aβ1-42 and p-Tau were increased;the mRNA expression and protein levels of AMPK was decreased;the activity of SOD,T-AOC,GSH-PX were decreased and content of MDA increased in plasma.However,compared with model group,the escape latency of MSC group and MSC+Tanshinone group rats was shorter,time percentage in the target quadrant and target quadrant frequency were markedly increased;moreover,the formation of senile plaque in the hippocampus was decreased;the number of surviving neurons in hippocampus was signifi cantly increased;the protein expression of Aβ1-42 and p-Tau were decreased;the mRNA expression and protein levels of AMPK was increased;the activity of SOD,T-AOC,GSH-PX were increased and content of MDA decreased in plasma.And MSC+tanshinone group was more significant than the
MSC group. Conclusion：Under the experimental conditions,MSC incubated with tanshinone ⅡA signifi cantly ameliorates spatial learning and memory impairment and reduce Aβ aggregation and tau protein phosphorylation levels induced by Aβ25-35 in rats,and its potential mechanism may be related
toanti-oxidative stress.     

Effects of Kaixin Jieyu Decoction (开心解郁汤) on Behavior and Glial Fibrillary Acidic Protein Expression in Cerebral Hippocampus of A Rat Vascular Depression Model

Objective:To explore the effects and anti-depression mechanisms of Kaixin Jieyu Decoction(开心解郁汤,KJD).Methods:The rat vascular depression(VD) model was established by ligation of bilateral common carotid arteries(LBCCA) combined with chronic unpredictable mild stress(CUMS).Forty Wistar rats were randomly divided into sham,VD model,VD + high-dose KJD[15.4 g/(kg·d) of crude drug],VD + medium-dose KJD[7.7 g/(kg·d) of crude drug],and VD + fluoxetine[2.4 mg/(kg·d)]groups(r=8 in each group),and the treatments lasted for 21 days.Changes of behavior and hippocampus pathology were observed.The level of glial fibrillary acidic protein(GFAP)protein and mRNA in hippocampus was detected respectively by immunohistochemistry and real-time polymerase chain reaction.Results:Compared with the sham group,rats in model group showed a variety of behavioral obstacles,including a significant reduction in sucrose consumption percentage,horizontal and vertical activity scores in open-field tests(P0.05 or P0.01),pathological damage like neuronal degeneration,necrosis,and a significant decrease of GFAP protein and mRNA in hippocampus(P0.01);compared with the model group,rats in the high-dose KJD group,medium-dose KJD group and fluoxetine group obtained notable higher behavioral scores,and pathological injury lessened in hippocampus with a increased expression of GFAP protein and mRNA(P0.05 or P0.01);compared with the medium-dose KJD group and fluoxetine group,GFAP mRNA in highdose KJD group expressed higer(P0.05).Conclusion:LBCCA combined with CUMS may cause depression-like behavioral changes resulting in the VD model of rats whose depression state can be ameliorated by KJD,and the mechanism of cerebral protection is related possibly with promoting expression of GFAP in hippocampus.