High glucose dialysate enhances peritoneal fibrosis through upregulating glucose transporters GLUT1 and SGLT1 / 浙江大学学报·医学版

To investigate the role of glucose transporter 1 (GLUT1) and sodium-glucose cotransporter 1 (SGLT1) in high glucose dialysate-induced peritoneal fibrosis.Thirty six male SD rats were randomly divided into 6 groups (6 in each):normal control group, sham operation group, peritoneal dialysis group (PD group), PD+phloretin group (PD+T group), PD+phlorizin group (PD+Z group), PD+phloretin+phlorizin group (PD+T+Z group). Rat model of uraemia was established using 5/6 nephrotomy, and 2.5% dextrose peritoneal dialysis solution was used in peritoneal dialysis. Peritoneal equilibration test was performed 24 h after dialysis to evaluate transport function of peritoneum in rats; HE staining was used to observe the morphology of peritoneal tissue; and immunohistochemistry was used to detect the expression of GLUT1, SGLT1, TGF-β1 and connective tissue growth factor (CTGF) in peritoneum. Human peritoneal microvascular endothelial cells (HPECs) were divided into 5 groups:normal control group, peritoneal dialysis group (PD group), PD+phloretin group (PD+T group), PD+phlorezin group (PD+Z group), and PD+phloretin+phlorezin group (PD+T+Z group). Real time PCR and Western blotting were used to detect mRNA and protein expressions of GLUT1, SGLT1, TGF-β1, CTGF in peritoneal membrane and HPECs., compared with sham operation group, rats in PD group had thickened peritoneum, higher ultrafiltration volume, and the mRNA and protein expressions of GLUT1, SGLT1, CTGF, TGF-β1 were significantly increased (all<0.05); compared with PD group, thickened peritoneum was attenuated, and the mRNA and protein expressions of GLUT1, SGLT1, CTGF, TGF-β1 were significantly decreased in PD+T, PD+Z and PD+T+Z groups (all<0.05). Pearson's correlation analysis showed that the expressions of GLUT1, SGLT1 in peritoneum were positively correlated with the expressions of TGF-β1 and CTGF (all<0.05)., the mRNA and protein expressions of GLUT1, SGLT1, TGF-β1, CTGF were significantly increased in HPECs of peritoneal dialysis group (all<0.05), and those in PD+T, PD+Z, and PD+T+Z groups were decreased (all<0.05). Pearson's correlation analysis showed that the expressions of GLUT1, SGLT1 in HPECs were positively correlated with the expressions of TGF-β1 and CTGF (all<0.05).High glucose peritoneal dialysis fluid may promote peritoneal fibrosis by upregulating the expressions of GLUT1 and SGLT1.

Influence of Butanol Extract from Stevia on the expression of single immunoglobulin interleukin-1 receptor-related molecule and nuclear factor kappa-B in depression mice liver / 中华实用儿科临床杂志

Objective To study the antidepressant-like effect of Butanol Extract from Stevia (BEFS) and its possible corresponding mechanisms.Methods Forty-five mice were randomly divided into control group,model group,model + Fluoxetine group,model + BEFS-3 g/kg group and model + BEFS-6 g/kg group.The chronic unpredictable mild stress(CUMS) for 3 weeks was used to make up depressive animal model.It was based on body weight,crossing numbers,rearing numbers and sucrose preference to assess whether the model was a success.After continuous lavage giving BEFS and fluoxetine for 2 weeks and subsequent behavioral test,the mRNA transcription of Single immunoglobulin interleukin-1 receptor-related molecule (SIGIRR) and nuclear factor kappa-B (NF-κB) in liver tissue of mice were detected by using quantitative real time-PCR.Results Three weeks after CUMS,behavior parameters were significantly decreased in mice exposed to CUMS compared with control group[(17.28 ±0.49) g vs (22.47 ± 0.82) g,P =0.000;(43.35 ± 1.68) scores vs (125.25 ± 3.07) scores,P =0.000;(3.92 ± 0.18) scores vs (15.92 ± 1.78) scores,P =0.000;(49.39 ± 2.05) ％ vs (67.18 ± 4.46) ％,P =0.001].However,2 weeks after dosing,body weight,crossing numbers,rearing numbers and sucrose preference were (19.03 ± 0.44) g,(70.50 ± 6.56) scores,(3.08 ± 0.77) scores and (41.27 ± 7.59) ％,respectively in model group.Compared to model group,behavior parameters of BEFS (3 and 6 g/kg BEFS) groups were significantly increased [BEFS-6 g/kg group:(24.42 ± 1.46) g,(99.75 ± 5.07) scores,(13.08 ± 1.60) scores,(64.31 ± 1.92) ％,all P ＜ 0.05;BEFS-3 g/kg group:(23.82 ± 1.72) g,(91.67 ± 5.69) scores,(6.92 ± 1.05) scores,(59.56 ± 5.31) ％,respectively,all P ＜ 0.05].Correspondingly,BEFS could reverse the mRNA expression of SIGIRR lowering and NF-κB rising in the liver of mice exposed to CUMS(BEFS-6 g/kg group:1.208 1 ± 0.102 2,1.437 7 ± 0.352 9;BEFS-3 g/kg group:0.768 0 ± 0.108 6,2.186 2 ± 0.203 8,all P ＜ 0.05).Conclusions BEFS can remarkably improve depression-like behaviors in CUMS mice and its antidepressant activity is mediated,at least in part,by upregulating SIGIRR and downregulating NF-κB in the liver.

Effect of rosiglitazone on the expression of AQP-1, VEGF-A and COX-2 in uremic rat of peritoneal dialysis / 中华肾脏病杂志

Objective To investigate the effect of rosiglitazone(RGZ) on peritoneal morphology,function and the expressions of Aquaporin 1 (AQP-1),vascular endothelial growth factor A (VEGF-A) and cyclooxygenase 2(COX-2) in uremic rat of peritoneal dialysis.Methods Thirty Sprague-Dawley rats were randomly divided into five groups.Group S (n=6) was subjected to sham operation.Group N (n=6) was subjected to nephrectomy with silicon catheter inserted,but no peritoneal exposure.Group P (n=6) was subjected to nephrectomy with silicon catheter inserted and receiving daily peritoneal injection through the catheter,using 4.25％ peritoneal dialysis fluid 10 ml twice a day for 2 weeks.Group R (n=6) was subjected to nephrectomy with silicon catheter inserted and receiving daily peritoneal injection through the catheter,using 4.25％ peritoneal dialysis fluid containing rosiglitazone (0.2 mg/kg) 10 ml twice a day for 2 weeks.Group GW (n=6) was subjected to nephrectomy with silicon catheter inserted and receiving daily peritoneal injection through the catheter,using 4.25％ peritoneal dialysis fluid containing rosiglitazone (0.2 mg/kg) and GW9662 (0.2 mg/kg) 10 ml twice a day for 2 weeks.After two weeks of dialysis,a 90 min peritoneal equilibration test was performed and the amount of ultrafiltration was accurately measured.The partial peritoneum tissues of rats were harvested and stained by hematoxylin-eosin (HE),then morphology changes of partial peritoneum were examined by light microscopy.The expression of AQP-1,VEGF-A and COX-2 in omentum were detected with immunohistochemistry assay.AQP-1,VEGF-A and COX-2 mRNA were detected by qRT-PCR.Results Morphology changes of partial peritoneum showed that compared with Group S,a dramatic increase in thickness of the mesothelium-to-muscle layer of peritoneum in Group N,P,R and GW(P ＜0.05).Compared with group P,the thickness significantly decreased in Group R(P ＜ 0.05).PET results showed that compared with Group S,ultrafiltration (UF) significantly reduced in Group P,R,and GW (P ＜ 0.05).Compared with Group P,ultrafiltration significantly increased in Group P,R,and GW (P ＜0.05).Compared with group S,the expressions of AQP1,VEGF-A and COX-2 mRNA and protein were significantly increased in group P,R and GW(P ＜ 0.05).Compared with group P,the expressions of AQP1,VEGF-A mRNA and protein were significantly decreased in Group R and GW(P ＜ 0.05).Compared with group P,the expressions of COX-2 mRNA and protein were significantly decreased in group R (P ＜ 0.05),while no differences in the expression of COX-2 mRNA and protein in group GW (P ＜ 0.05).Conclusions Rosiglitazone can inhibit peritoneal interstitial and vascular proliferation,protect peritoneal function and increase ultrafiltration.Rosiglitazone can protect peritoneal function probably by inhibiting expression of VEGF-A and COX-2.

The role of adiponectin postconditioning in myocardial ischemia reperfusion injury in 3-week old rats / 中华实用儿科临床杂志

Objective To study the role of adiponectin (ADP)post-conditioning in alleviating myocardial ischemia reperfusion injury (MIRI) in 3-week old rats.Methods SD rats (3-week old) were randomly divided into 4 groups by random digits table (n =6):sham group was processed identically except that the left anterior descending coronary artery(LAD) was not ligated;MIRI group was subjected to occlusion of the LAD followed by reperfusion,occlusion for 30 min and reperfusion for 120 min;ADP group was subjected to injection of ADP into femoral vein before reperfusion;LY294002 group (ADP post-conditioning and LY294002) was subjected to injection of ADP and LY294002 into femoral vein before reperfusion.The titer of lactate dehydrogenase(LDH),cardiac troponin I(cTnl) and malondialdehyde(MDA) in plasma were observed.The levels of superoxide dismutase (SOD)and MDA in myocardial tissue were observed.Results The levels of LDH,cTnI and MDA in plasma of MIRI group was (732.11 ± 111.29) IU/L,(38.05 ± 6.17) g/L and (4.49 ± 0.17) nmol/L,respectively,compared with sham group,the levels of LDH,cTnI and MDA in plasma of MIRI group increased obviously(all P ＜ 0.05);the levels of LDH,cTnI and MDA in plasma of ADP group were (581.72 ± 68.41) IU/L,(20.70 ± 5.43) g/L and (3.10 ± 0.20) nmol/L,respectively;compared with MIRI group,the levels of LDH,cTnⅠ and MDA in plasma of ADP group declined significantly (all P ＜0.05);the levels of LDH,cTnI and MDA in plasma of LY294002 group were (701.10 ±99.59) IU/L,(33.13 ± 4.32) g/L and (4.19 ± 0.46) nmol/L,compared with ADP group,the levels of LDH,cTnI and MDA in plasma of group LY294002 increased remarkably (t =2.477,1.804,2.961,all P ＜ 0.05).Compared with sham group,the SOD level decreased and the MDA level increased in myocardial tissue of MIRI group(all P ＜ 0.05);compared with MIRI group,the SOD level increased and the MDA level decreased in myocardial tissue of ADP group (all P ＜ 0.05);compared with ADP group,the SOD level decreased and the MDA level increased in myocardial tissue of LY294002 group (all P ＜ 0.05).Conclusions ADP postconditioning is perhaps a protective factor for alleviating myocardial ischemia reperfusion injury in 3-week-old rats,the protective effect is perherps related to alleviating the damage of the lipid peroxidation by signaling pathway of adiponectin/phosphoinositide-3 kinase/protein kinase B.

Effect of rosiglitazone on expression of AQP1, VEGF-A and COX-2 in human peritoneal microvascular endothelial cells / 中国病理生理杂志

AIM:To investigate the effect of rosiglitazone on the expression of aquaporin-1 (AQP1), vascular endothelial growth factor-A ( VEGF-A ) and cyclooxygenase-2 ( COX-2 ) in human peritoneal microvascular endothelial cells.METHODS: Cultured peritoneal microvascular endothelial cells were divided into 4 groups.The morphological changes of the cells were observed under inverted microscope .The protein expression of AQP1, VEGF-A and COX-2 in hu-man peritoneal microvascular endothelial cells was determined by Western blotting .The mRNA expression of AQP1, VEGF-A and COX-2 in the cells was measured by real-time PCR.RESULTS:Rosiglitazone stimulated the proliferation of the cells.Rosiglitazone up-regulated the expression of AQP1, and down-regulated the expression of VEGF-2 and COX-2 at mRNA and protein levels in the cells .The PPAR-γantagonist GW9662 partly inhibited the up-regulation of AQP1 expres-sion by rosiglitazone (P0.05).CON-CLUSION:Rosiglitazone up-regulates the expression of AQP 1 and down-regulates the expression of VEGF-A and COX-2 in human peritoneal microvascular endothelial cells , thus promoting water transportation and attenuating peritoneal fibrosis during peritoneal dialysis .

Modified method of chromosome G-banding in human peripheral blood / 中国组织工程研究

BACKGROUND: Routine methods of chromosome G-bending are complicated, time-consuming and worse in effect, which is not suitable for clinical examination and teaching investigation of chromosomal disorder.OBJECTIVE: To seek for proper method of improving the effect of chromosome G-banding.DESIGN: Observation experiment.SETTING: Xinxiang Medical University.MATERIALS: The experiment was conducted in the Laboratory of Morphous, Xinxiang Medical University between January 2001 and January 2005. 376 blood samples were obtained from patients with infertility and sterility or those had abnormal childbearing history, who came from the Clinic of Gynecology and Obstetrics, Third Hospital Affiliated to Xinxiang Medical College between 2001 and 2004. Main agents: RPMI1640 culture fluid (GIBCO Co., Ltd), 20% calf serum (CS), colchicines, 0.075 mol/L KCL, methanol, glacial acetic acid, trypase (SIGMA Co., Ltd) and Giemsa staining solution.METHODS: Modified method of human peripheral chromosomal preparation and G-banding: The procedures were the same as routine methods, while partial influential factors were regulated, for example, the action time of colchicines was set at 2 hours before the ending; 1 mL of fixation fluid (the ratio of methanol and glacial acetic acid was 2:1) was added for pre-fixation, and samples were mixed and centrifuged at 1 500 r/minute for 10 minutes. After re-centrifugalization,fresh fixation fluid was added to prepare for cell suspension and glass slide by according to the amount of cells.Above-mentioned glass samples were baked at 50 ℃ for 1-2 hours, naturally cooled to 37 ℃, digested for 3-5 minutes by immersing into trypase, rapidly stained for 10 minutes with Giemsa staining solution, and counted by the test under microscope to observe 3 000 metaphases, and the percentages; The percentage of 400-600 metaphases and well-dispersion rate were calculated. Well-dispersion rate of chromosome referred to the overlapping after dispersion with complete numbers. The trabant and kinetic body were obvious and in bright color with clear shape. Moreover, all chromosomes were on the same plane. The percentage of metaphase = the number of metaphase cells (400-600)/number of cells under observation×100%.MAIN OUTCOME MEASURES: The percentage of 400-600 pieces of metaphases in samples and good integration chromosome.RESULTS: 400-600 metaphases obtained with routine methods accounted for 52% with a well-dispersion rate of 68%,while 400-600 metaphases obtained with modified methods accounted for 75% with a well-dispersion rate of 86%, and there were significant differences between the two groups (P ＜ 0.05).the stripes are chear.CONCLUSION:Chromoseme metaphases obtained with modified methods are more in number and good in dispersion with proper size. Giemsa shows that the stripes art chear.

Effect of eggembryosin on cosmetology, anti-senility and immunoloregulation / 中国组织工程研究

BACKGROUND: Eggembryosin is characterized by tonifying kidney and vital essence, invigorating qi and spleen, replenishing and activating blood, beautifying and nourishing face, and improving constitution of whole organism.OBJECTIVE: To observe the influence of eggembryosin on aging index,quantity of hemocytes and quality of immune organs of animal models, and analyze its effect on cosmetology, anti-senility and immunoloregulation.DESIGN: Randomized controlled observation.SETTING: Department of Pharmacology, Xinxiang Medical College.MATERIALS: The experiment was carried out in the Department of Pharmacology of Xinxiang Medical College between April 1998 and September 2002. A total of 80 Kunming mice of both sexes were selected in this study.METHODS: ① Effect of eggembryosin on activity of superoxide dismutase (SOD) in serum, content of malondialdehyde (MDA), lipofuscin and hydroxyproline respectively in serum, liver and tendon of tail of aging mice: Thirty mice were divided into 3 groups according to randomly lined table, including eggembryosin group, D-galactose model group and normal control group with 10 in each group. The former two groups were injected with D-galactose to copy aging models, and eggembryosin group were perfused with eggembryosin to measure activity of SOD in serum, content of MDA, lipofuscin (aging index) and hydroxyproline (elasric index of skin) respectively in serum, liver and tendon of tail. ② Effect of eggembryosin on mass of immune organs of mice with low immunological function: Thirty mice were selected and divided as the same way mentioned above. Mice in eggembryosin group were perfused with eggembryosin. In addition, those in eggembryosin group and cyclophosphamide (CAP) group were peritoneally injected with CAP to copy immunosuppression models and calculate index of thymus (spleen) (mg/g) = weight of thymus (spleen)/body. ③ Effect of eggembryosin on quantities of erythrocytes and hemoglobin of mice with blood deficiency: Twenty mice were divided into 2 groups according to randomly lined table, including eggembryosin group and normal control group with 10 in each group. Values of erythrocytes and hemoglobins of mice were measured before administration. Models in types of blood-deficiency blood-loss were copied with tail xsanguinations, and then, mice were perfused with eggembryosin and saline, respectively. Blood of mice were collected from their tails to measure values of erythrocytes and hemoglobin.MAIN OUTCOME MEASURES: Activity of superoxide dismutase (SOD) in serum, content of malondialdehyde (MDA), lipofuscin and hydroxyproline respectively in serum, liver and tendon of tail of aging mice, index of thymus, and spleen, contents of blood erythrocytes and hemoglobins.RESULTS: A total of 80 mice were involved in the final analysis without any loss. ① Activity of SOD in serum and contents of hydroxyproline in tendon of tail of mice in eggembryosin group were significantly higher than those of mice in D-galactose model group (P ＜ 0.01); content of MDA in serum and content of lipofuscin in liver of mice in eggembryosin group were significantly lower than those of mice in D-galactose model group (P＜ 0.01). ② Indexes of thymus and spleen of mice in eggembryosin group were higher than those of mice in CAP group (P ＜ 0.05-0.01). ③ Increasing values of blood erythrocytes and hemoglobin of mice in eggembryosin group were significantly higher than those of mice in normal control group (P ＜ 0.01).CONCLUSION: Eggembryosin taken orally can improveanti-oxidation of organism and elasticity of skin, increase quality of immune organs damaged by CAP and immunological function, and nourish the blood.

Effects of Huperzine A on the memory and learning behavior in D galactose induced senile mice / 中国组织工程研究

BACKGROUND:Huperzine A is a reversible choline enzyme(CHE) inhibitor initially developed in China,which has favorable therapeutic effectiveness in benign memory dysfunction,and could significantly promote the learning process and memory reappearance in rodent animals. OBJECTIVE:To comprehend the impacts of Huperzine A on the enhancement of learning, memory and behavior in platform and maze experiments in D Galoctose(Gal) induced senile mouse model. DESIGN:A randomized and controlled trial. SETTING:Institute of Pharmaceuticals of Xinxiang Medical College. MATERIALS:Study was performed in the Laboratory of Physiology and the Institute of Pharmaceuticals of Xinxiang Medical College from January to December in 2002.A total of 100 healthy Kunming mice were selected for platform and electric maze experiments with 50 mice in each experiment.Huperzine A tablet was obtained from Henan Zhulin Zhongsheng Pharmaceutical Company Limited(batch number:031001).D Gal is the product of Shanghai No.2 Reagent Factory. MAIN OUTCOME MEASURES:The latency in the safe area on the platform and the times of electric shock occurred within 5 minutes were observed in mice in platform experiment;the times of the mice mistakenly escaping to the right electric attacking area were observed in mice in electric maze experiment. CONCLUSION:Huperzine A can effectively improve the learning and memory ability of D Gal induced senile mice, and there is dose effect relationship within certain range. Qiu PY,Chen ZY,Wang YX,Zhang JX,Wu ZZ.Effects of Huperzine A on the memory and behavior in D galactose induced senile mice.Zhongguo Linchuang Kangfu 2005;9(4):247- 9(China) [www.zglckf.com]

Effect of cristata L flavonoid on expression of bone morphogenetic protein and function of tubular reabsorption of rats with diabetes mellitus / 中国组织工程研究

BACKGROUND: Cristata L flavonoid is a kind of plant estrin, which possesses multiple physiological function, has no toxicity and adverse effect, and is effective in treating and preventing osteoporosis.OBJECTIVE: To study the effect of cristata L flavonoid on bone morphogenetic protein, urine inorganic salt and content of lysozyme of rats with diabetes mellitus (DM).DESIGN: Randomized grouping design and controlled study.SETTING: Pharmaceutical Laboratory of Xinxiang Medical College.MATERIALS: The experiment was completed in the Xinxiang Medical College from September to December 2003. Totally 24 health male SD rats were randomly divided into three groups: normal control group, diabetes mellitus (DM) group, DM + cristata L flavonoid group with 8 in each group.were injected intraperineally with 60 mg/kg streptozotocin, and 48-72 hours later, blood of rear caudal vein was collected to measure total blood glucose.Rats were determined as DM if the blood glucose was ≥ 16.7 mmol/L; oth erwise, 60 mg/kg streptozotocin was injected once more. After modeling,cristata L flavonoid and rats in normal control group were given the same was measured with atomic absorbency method and content of urine sodium histochemistry of bone morphogenic protein-2 (BMP2) in bone was detected with streptavidin tagged by peroxidase and immunohistochemic expression lysozyme reflected reabsorption function of renal tubule was measured with with t-test. MAIN OUTCOME MEASURES: Comparison between content of calcium, sodium, kalium in urine and lysozyme and immunohistochemic expression of BMP2 10-week intervention later.calcium and sodium in urine in DM model group were obviously higher than those in normal control group, but content of kalium in urine was obviously lower (P ＜ 0.05-0.01). Contents of calcium in urine in DM +cristata L flavonoid group were obvioualy lower than those in DM model group and normal control group were obviously higher than those in DM model group, and content of lysozyme in urine in those groups was obviously lower than that in DM model group (P ＜ 0.05-0.01).CONCLUSION: Expression of BMP2 of DM rats is decreased, but after supplement of cristata L flavonoid compound, the expression is increased.In addition, output of urine calcium and sodium in DM rats is decreased,and reabsorption function of renal tubule is increased.