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Objective To explore the protective effects of schisandrin B (Sch B) on hypoxia injury induced by cobaltous chloride (CoCl2) in human proximal renal tubular epithelial (HK-2) cells, and the possible mechanism thereof. Methods HK-2 cells were randomly assigned to four groups:control group (Con, cells were untreated), CoCl2 group (CoCl2, cells were treated with 600μmol/L CoCl2 for 24 h), Sch B pretreat group (CoCl2+Sch B, cells were pretreated with 1μmol/L and 10μmol/L Sch B for 2 h) and Sch B group (Sch B, cells were treated with 1μmol/L and 10μmol/L Sch B for 2 h). CCK-8 kit was used to detect the cell viability of four groups. Flow cytometry was used to detect the apoptotic rate of four groups. The protein expression of hypoxia-inducible factor 1α(HIF-1α) was assessed by Western blot assay. The expressions of HIF-1α and inducible nitric oxide synthase (iNOS) mRNA were determined by RT-PCR. Results Compared with the control group, after treated with 600 μmol/L CoCl2, the cell viability was decreased, and the apoptosis was increased, the expressions of HIF-1α and iNOS mRNA were up-regulated in HK-2 cells. There was no significant difference in the expression of HIF-1α mRNA between control group and CoCl2 group. Compared with the CoCl2 group, after pretreated with 1μmol/L and 10μmol/L Sch B, the cell viability was increased and the apoptosis was decreased, the expressions of HIF-1α and iNOS were down-regulated in HK-2 cells. There were no significant differences in the cell viability and apoptotic rate between control group and Sch B group. Conclusion Pretreatment with Sch B can reduce the apoptosis of HK-2 cells by inhibiting the expression of HIF-1α and iNOS mRNA, which shows protective effects on hypoxia injury.
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Objective To observe the intervention effect of Schisandrin B (Sch B) on cisplatin induced acute kidney injury (AKI) in mice and its possible mechanism.Methods Twenty-five BALB/c mice were randomly divided into blank control group, model group, low and high dose of Sch B intervention groups and Sch B control group. Olive oil with Sch B was administered by gavage at the dose of 20 mg/kg or 100 mg/kg for low and high dose of Sch B intervention groups respectively; olive oil with Sch B 100 mg/kg was applied by gavage to the Sch B control group; the same volume of olive oil was perfused into the gastric cavity in the blank control group and model group; the above measures in various groups were consecutively used for 5 days. On the 3rd day of the experiment, AKI mice model was established by intraperitoneal injection of cisplatin (20 mg/kg) once and the same measure was given to the low and high dose of Sch B intervention groups; 1 mL/kg normal saline was injected into the peritoneal cavity in the bland control group and Sch B control group. At the end of the experiment, the serum creatinine (SCr) level was determined; apoptosis of renal tubular epithelial cells were detected by using terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay; the morphological changes of renal tubular epithelial cells were observed by hematoxylin eosin (HE) staining, and renal tubular injury score was evaluated; p53 protein content in the kidney tissue was measured by immunohistochemical analysis; furthermore, expressional level of p53 protein in renal tissue was tested by Western Blot.Results Compared with the blank control group, the level of SCr (μmol/L: 86.77±10.97 vs. 14.37±0.81), renal tubular injury score (9.67±1.20 vs. 1.00±0.45), the count of apoptotic renal tubular epithelial cells (cells/200 power field: 20.00±2.13 vs. 2.30±0.40) in the model group were all increased (P < 0.05 orP < 0.01), and p53 protein content (cells/400 power field: 13.40±2.66 vs. 57.30±3.82), and the expression of p53 protein [absorbency (A value) ratio: 0.79±0.09 vs. 1.42±0.09] in model group were decreased (bothP < 0.01). Compared with the model group, in the low and high dose Sch B intervented groups, the level of SCr (μmol/L: 21.98±5.52 and 37.45±5.04), renal tubular injury score (5.67±0.76 and 6.17±0.65), the count of apoptotic renal tubular epithelial cells (cells/200 power field: 10.60±1.05 and 11.60±1.45) were all reduced (allP < 0.01), p53 protein content (cells/400 power field: 42.40±3.67 and 45.90±2.31) and the expression of p53 protein (A value ratio: 1.36±0.16 and 1.25±0.11) were increased (bothP < 0.01). HE staining showed the pathological changes of renal tubules, such as renal tubular epithelial cellular fusion, vacuolization, cast formation, and tubular lumen constriction/dilation in model group; the pathological changes in kidney tissues observed in low and high dose Sch B intervention groups were milder than those in model group.Conclusion Sch B plays a beneficial role in the cisplatin induced AKI in mice, and its protective effect might be mediated by decreasing SCr, regulating p53 protein expression level and inhibiting the apoptosis of renal tubular epithelial cells.
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Objective To investigate the therapeutic effect of traditional Chinese medicine(TCM)recipe of nourishing kidney and activating blood for treatment of cisplatin induced acute kidney injury(AKI)in mouse model. Methods Twenty-one male BALB/c mice were randomly divided into three groups:control group,AKI model group and treatment group with the above TCM recipe(each n=7). The AKI model was reproduced by a single intraperitoneal injection of cisplatin 20 mg/kg;the TCM recipe with dosage of 10μL · g-1 · d-1 was given to the treatment group,while equal volume of normal saline was given to the model and control groups by gavage,lasting for 6 days in all groups. The changes of each mouse body weight were observed;mouse venous blood serum creatinine(SCr)level was detected,changes of renal tissue weight and its histopathology were observed under light microscope,and the score of acute tubular necrosis(ATN)was calculated. Results Compared to the control group,the body and renal weight in AKI model group were significantly lowered〔body weight(g):17.18±0.29 vs. 19.33±1.43,renal weight(g):0.28±0.01 vs. 0.32±0.11,both P<0.01〕,and SCr and ATN scores were significantly increased in AKI model group〔SCr(μmol/L):86.77±10.97 vs. 14.37±0.81,ATN score:3.33±0.52 vs. 0.17±0.41,both P<0.01〕. Compared to AKI model group,the body weight(g:18.70±0.28)and renal weight(g:0.31±0.01)in the treatment group were markedly increased,and SCr(μmol/L:21.98±5.52)and ATN scores(2.00±0.63)were significantly lower than those of the AKI model group(all P<0.01). Under optical microscope,there were mouse renal tubular epithelial cell edema and necrosis,renal interstitial inflammatory cell infiltration,the glomerular form roughly normal in the AKI model group;compared with the AKI model group,in the treatment group the degree of mouse renal tubular necrosis was significantly reduced and glomerular form was basically normal. Conclusion The TCM recipe of nourishing kidney and activating blood can reduce SCr and improve the pathological changes of renal tissue in cisplatin induced AKI mouse models,thus it has therapeutic effect for treatment of AKI in mice.
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Objective To explore the effect of schisandra chinensis fruit ethanol extract on nephrin and desmin ex-pression in adriamycin(ADR) induced podocyte injury in vitro. Methods Conditionally immortalized mouse podocytes were treated with ADR for 24 h in vitro, then the medium was changed to medium with SE(250 mg/L)for 24 h. Podocytes were di-vided into four groups:control group,model group, SE intervention group and SE group. The expression of nephrin in podo-cytes was detected by immunofluorescence. Western Blot was employed to assess nephrin and desmin expression. Transcrip-tion level of nephrin and desmin were determined by qRT-PCR. Results Nephrin expression was distributed along the cell membrane in linear or granular pattern in control group and SE group. Fluorescence intensity in model group was lower than that of control group SE group and SE intervention group. There was no significant difference of nephrin and desmin protein and mRNA level between control group and SE group. Compared with the model group, protein and mRNA level of nephrin was lower than that of control group and SE intervention group. The protein expression and mRNA transcription of desmin in model group was higher than those in control group and SE intervention group (P＜0.05). Conclusion SE(250 mg/L)has no harmful effect on the podocytes in vitro. SE can protect the podocytes from damage by adriamycin in vitro. SE not only up-regulate the expression of nephrin, but also down-regulate of desmin expression.
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Objective To study the change law between deficiency of kidney-Yang and stagnation of blood, and supply a study evidence for the therapy method of invigorate the kidney and promoting blood flow. Method 37 Wistar rats which were made of the model of deficiency of kidney-Yang, were randomly divided into three groups:control group, model group and treatment group. Changes including appearance, general objective sign, organ weight, microcirculation and blood rheology were recorded. Result Phenomenons, including rare fur, depression, lazy, duck stools and purpl dim in micrangium micrangium, occurred in model group. The weight of adrenal body and seminal vesicle and the contents of serum andrusol in model group were lower than those in control group. And in model group, significant increasing TC and LDL-C and decreasing HCL-C were observed. Conclusion Deficiency of kidney-Yang is associated with stagnation of blood, and the time delay of stagnation of blood can lead deficiency of kidney-Yang.
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Objective To detect the risk factors for type 2 diabetic nephropathy(DN).Methods A case-control study was conducted,136 hospitalized DN patients and 152 hospitalized type 2 diabetic patients without DN as control group were enrolled in the study.Information including demographic characteristics,history of diabetes and diabetic nephropathy,behaviors,social psychological characters,clinical data and laboratory data was collected retrospectively.Statistic methods including ?2 test,u test and non-conditional logistic regression were conducted to detect the risk factors of DN.Results Between the cases and the controls,there were significant differences in the course of diabetes mellitus(DM),being complicated with hypertension,hyperlipidemia,coronary heart disease,diabetic retinopathy,diabetic foot,diabetic peripheral neuropathy,pulse pressure,laboratory features as tiglyceride,total cholesterol,creatinine,uric acid,and behaviors including control of systolic blood pressure,exercise after DM.Results of multivariate logistic regression analysis showed that course of DM(OR=2.48 95%CI:1.87～3.42),being complicated with hypertension(OR=1.38 95%CI:1.09～1.62) and blood creatinine level(OR=6.23 95%CI:3.03～9.82) were risk factors for DN.Conclusion Longer course of DM,with hypertension and high level of blood creatinine were the risk factors of DN.DM patients should strictly control their blood pressure and blood sugar,periodically inspect renal function and blood creatinine,and correct dyslipidemia to prevent the development and progression of DN.
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Objective:To investigate the expression of mmp-2 in mesangial proliferative glomerulonephritis(MsPGN) rat renal tissue and the effects of nourishing kidney and activating blood flow recipe of TCM. Methods: 54 Male SD rats were divided into control group, MsPGN group and treatment group with nourishing kidney and activating blood flow recipe of TCM. Immunohistochemical staining, flow cytometry, western blot and RT-PCR were used to check the protein expression of mmp-2. Results: In MsPGN group, following with the disease progressing, the glomerulus grew graduatly, ECM increased, the basement membrane of glomerulus was thicker. The immunohistochemical staining showed that the chief positive granules were deposited in glomerulus and renal tubular, the expression of mmp-2 was lower than that in control group, but in treatment group the expression of mmp-2 was higher than that in MsPGN group. Conclusion: In MsPGN renal tissue, the expression of mmp-2 was reduced, its activity was weakened. Nourishing kidney and activating blood flow recipe of TCM could induce the expression of MMP-2 and partly recover the activity. Nourishing kidney and activating blood flow recipe of TCM could resist glomerular sclerosis and postpone the course of chronic renal failure.
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Objective:To investigate the expression of matrix metalloproteinase(MMP) and tissue inhibitor of matrix metalloproteinase(TIMP)in mesangial proliferative glomerulonephritis(MsPGN) rat renal tissue and the effects of nourishing kidney and activing blood recipe of traditional Chinese medicine.Methods:Chose 54 male rats and separated them into control group, MsPGN group and treated group with nourishing kidney and activing blood recipe of traditional Chinese medicine.Chose six rats per group at the time of the second,the fourth and the eighth week.The sections were treated with HE and PAS stains and the routine pathologic study was carried.Immunohistochemical staining was used to check the expression of MMP-2,TIMP-2, membrance type metalloproteinases-1(MT1-MMP) and transforming growth factor-?1(TGF-?1).Flow Cytometry was used to check the protein expression of MMP-2,TIMP-2,MT1-MMP and TGF-?1 in rat renal tissue.Results:In MsPGN group,following with the disease progressing,the glomerulus grew graduatly,ECM became more and more,the basement membrane of glomerulus was thicker.The immunohistochemical staining showed the expression of MMP-2 and MT1-MMP was lower compared with control group,but in treated group the level was weaker,the expression of TIMP-2 and TGF-?1 was higher compared with control group,but in treated group the level was weaker too. At the fourth and the eighth week, the Flow Cytometry showed that in MsPGN group the FI of MMP-2 was lower compared with control group(P
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Objective To investigate the mechanism of Bushen Huoxue Recipe (Recipe to reinforce the kidney and activate blood) in the treatment of glomerulosclerosis of the rats with mesangial proliferative glomerulonephritis (MsPGN).Methods Fifty-four SD male rats were randomized into normal group, model group, and treated group with 18 rats in each. The MsPGN models were made by injection of anti-rat-thymocyte antiserum (ATS) through the tail vein. After modeling, the treated group was administered Bushen Huoxue Recipe with a dosage of 5ml/kg, once a day. Six rats from each group were selected in the second, fourth, and eighth week respectively to determine the content of urine protein (Upro), blood urea nitrogen (BUN), and serum creatinine (SCr), and the kidney was cut for pathological observation and immunohistochemical detection of the kidney tissues, the expression of Ⅳ-type collagen (Col-Ⅳ) and laminin (LN).Results The pathological determination showed that, at the same time points, the hyperplasia of mesangial cells in the treated group was significantly relieved and mesangial matrix reduced as compared with the model group. The content of Upro, BUN and SCr at all time points in the treated group was significantly decreased as compared with the model group (P