4.Establishment of a new rat model of chronic cyclosporine A nephrotoxicity.
Qiao-ling SUN ; Yi-pu CHEN ; Hong-liang RUI
Acta Academiae Medicinae Sinicae 2010;32(2):205-209
OBJECTIVETo establish a new rat model of chronic cyclosporine A nephrotoxicity and explore its features.
METHODSTotally 24 male SD rats were equally randomized divided into 3 groups: sham-adrenalectomized (sham-ADX) group, ADX group and ADX plus cyclosporine A (CsA) group. Rats in ADX and CsA group first underwent adrenalectomy, followed by the administration of placebo or dexamethasone, respectively. Rats in sham-ADX group received sham adrenalectomy and distilled water as control. Six weeks later, all rats were sacrificed and the following indicators were evaluated: urine protein excretion, creatinine clearance, aldosterone level in serum and urine, aldosterone level and its synthase CYP11B2 gene expression in kidney, serum natrium and potassium, urine natrium and potassium excretion, and tubulointerstitial fibrosis by masson trichrome stain.
RESULTSIn ADX and CsA group, serum and urine aldosterone were undetectable on the second post-operative day, with other observations including natriuresis, hyponatremia, decreased urine potassium excretion, and hyperpotassemia, suggesting that adrenals were removed intact and the adrenalectomy was successful. Rats in CsA group showed increased urine protein, decreased creatinine clearance and tubulointerstitial fibrosis, suggesting that a model of chronic CsA nephrotoxicity was successfully established. At the endpoint, serum potassium, serum aldosterone, urine potassium and urine aldosterone excretion partially retrieved. Natrium in serum and urine was not significant different between ADX group/CsA group and sham-ADX group. Local renal aldosterone and its gene expression were remarkably upregulated.
CONCLUSIONSWe successfully established a new rat model of chronic CsA nephrotoxicity by adrenalectomy without low sodium diet. After adrenalectomy, local renal aldosterone in kidney may compensate for circulatory aldosterone deficit to maintain electrolyte balance.
Acute Kidney Injury ; chemically induced ; Adrenalectomy ; Aldosterone ; metabolism ; Animals ; Cyclosporine ; toxicity ; Disease Models, Animal ; Immunosuppressive Agents ; toxicity ; Kidney ; drug effects ; metabolism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley
5.Complications of Hyperbaric Treatment
Yiping TANG ; Wei XU ; Dengli FU ; Yi CAO ; Qi LING ; Liang SUN
Journal of Kunming Medical University 2001;22(1):89-91
To analyze complications of hyperbar ic treatment and the management of these complications. Methods: There are 129 cases treated with huperbaria, 125 were severe head injury, 2 were brain infarction, one was carbon monoxide toxication and an other was anesthesia accident. These complications were analyzed. Results: There were 7 of 129 cases suffered with headache and vertigo, one of them was dead. Conclusion: Severe h ead injury patients treated with hyperbaria must be careful, cease the huperbaric treatmen t immediately whenever patients feel discomfort.
6.Effects of tidal volume with different amount of gas after recruitment maneuver ventilation on lung vascular endothelial diastole function in rats with acute lung injury
Jianqiang WANG ; Chun PAN ; Ling LIU ; Liang JIN ; Yi YANG ; Guomin LI ; Dong YUAN ; Haibo QIU
Chinese Journal of Emergency Medicine 2010;19(9):931-937
Objective To observe the effects of recruitment maneuver (RM) and tidal volume with different amount of gas after RM ventilation on lung diastole function in rats with acute lung injury (ALI). Method ALI rat models were induced by intravenous infusion of lipopolysaccharide (LPS) in dose of 6 mg/kg. Twenty-five rats were randomly(random number) divided into control group ( n = 5), ALI group ( n = 5), low tidal volume group (LV group,VT= 6 mL/kg, n = 5), sustained inflation (SI) with low tidal volume (SI+ LV group, VT=6 mL/kg, n = 5), and SI with moderate tidal volume group (SI+ MV group, VT= 12 mL/kg, n = 5). The RM carried out by using SI with airway pressure 30 cmH-2O for 30 seconds, and the positive end-expiratory pressure (PEEP)was set at 5 cmH2O. Lung tissue was taken after mechanical ventilation for 5 hours. The mean arterial pressure (MAP) was monitored throughout the entire course of experiment. Endothelin-1 ( ET-1 ), endothelial nitricoxide synthase (eNOS), and acetylcholine-(Ach-) induced endothelium-dependent relaxation response of isolated pulmonary artery rings were investigated after mechanical ventilation for 5 hours. Results The LPS increased the ET-1 level in lung tissue, decreased the level of eNOS in lung tissue, and impaired the Ach-induced endothelium-dependent relaxation response in pulmonary vassals, without obvious influence on systemic hemodynamics. SI + LV significantly reduced LPS-induced elevation of ET-1 level, and increased the level of eNOS, and significantly lessened endothelial dysfunction and ameliorated dysfunction od endothelium-dependent relaxation in pulmonary vas sals. Conclusions RM with high tidal volume or lowtidal volume ventilation could improve the lung vascular endothelial function of rats with acute lung injury, and RM with low tidal volume ventilation could lessen more the injury of lung vascular endothelial diastole function in rats with acute lung injury.
7.Immunoregulatory effects of bone marrow-derived mesenchymal stem cells from NZBWF1/J mice on T lymphocyte subsets balance
Hui LI ; Jun LIANG ; Ya-Yi HOU ; Le-Ming FAN ; Ling-Yun SUN ;
Chinese Journal of Rheumatology 2003;0(08):-
Objective To investigate the immunoregulatory effect and compare the different regula- tions of bone marrow mescnchymal stem cells(MSCs)derived from both lupus(NZBWF1/J)and normal(BALB/ c)mice on T lymphocytes in vitro.Methods MSCs from NZBWF1/J and BALB/c mice bone marrow were iso- lated and expanded,and identified by the surface phenotypes.CD3~+ T lymphocytes isolated by nylon wool columns were stimulated by phorbol myristate acetate(PMA)and co-cultured with or without the two strains of MSCs for 24 h.Intracellular eytokines of T cell,such as interferon(IFN)-?,interleukin(IL)-4,IL-12,IL-6, were analyzed by flow cytometry and quantification of transcription factors T-box expressed in T cells(T-bet) and GATA-binding protein 3(GATA-3)were detected by reverse transcriptase PCR(RT-PCR).T cell apop- tosis was assessed by flow cytometry using rhodamine123.Results The results showed that a decrease of CD3~+ T cell apoptosis was seen when NZBWF1/J MSCs or BALB/c MSCs were added to T cells stimulated by PMA(P<0.05),and an increase of TH2 cytokines by NZBWF1/J MSCs and TH1 eytokines by BALB/c MSCs were observed in the CD3~+ T cells eo-cuhured with MSCs(P<0.05).Conclusion It is suggested that the al- teration of T subsets caused by MSCs may interfere with the systemic lupus erythematosus(SLE)development and normal MSCs may be effective in the improvement of SLE.NZBWF1/J MSCs have defective immunoregula- tory function when compared with MSCs from healthy mouse strains.
8.Effects of carbon nanoparticles-epirubicin suspension on cell proliferation and apoptosis in breast cancer MCF-7 cells
Yongsong ZHANG ; Quankun LIANG ; Ling ZHONG ; Xiaowei QI ; Beige ZONG ; Junze DU ; Yi ZHANG ; Linjun FAN
Chinese Journal of Endocrine Surgery 2017;11(2):110-114
Objective To explore the effect of carbon nanoparticles-epirubicin suspension on proliferation and apoptosis in human breast cancer MCF-7 cells.Methods MCF-7 cells were cultured with different concentrations of CNP-EPI in vitro.CCK-8 assay was used for determinate inhibition effect of CNP-EPI on the proliferation of MCF-7 cells at different concentration and different time.According to the determination of IC90,5 μg/ml CNP-EPI was selected,and cell morphology and cell apoptosis rates were observed after 24 h.Results The inhibition effect of the CNP-EPI was stronger significantly in CNP-EPI group than in normal control group within 24 h,48 h,72 h when the concentration was from 1 μg/ml to 200 μg/ml (P<0.01).The inhibition of CNP-EPI on the proliferation of MCF-7 cells was gradually strengthened in a dose-dependent relation within the same time,and the inhibition effect is reduced in the same concentration of drugs with the time extension,but it still has a strong inhibitory effect in 72 h,and the inhibition effect of different concentration of CNP was not obvious on MCF-7 cells.Obvious changes of cell morphology were observed under inverted microscope such as:a lot of dead cells,cell adherent growth poor,cell morphology became round and karyopycnosis etc,in 5 μg/ml CNP-EPI group after 24 h.However,no obvious abnormity of cell norphology was observed in normal control group and corresponding CNP group.Late apoptosis rate was (14.57±2.41) %,the mortality rate could reach (78.63±-20.55)% in 5 μg/ml CNP-EPI group after 24 h.The mortality rate and apoptosis rate of cells was higher significantly in CNP-EPI group than in CNP group and normal control group (P<0.05).Conclusion CNP-EPI can obviously inhibit the proliferation or kill human breast cancer MCF-7cells,and the inhibition effect of CNP-EPI on proliferation of breast cancer cells might be the result of delayed releasing of EPI.
9.The control effects of FLT3 signaling-dependent pulmonary conventional dendritic ceils on the initiation of acute lung inflammation response to lipopolysaccharide induced acute lung injury in mice
Liang DONG ; Hongli HE ; Jun LIU ; Ling LIU ; Yi YANG ; Haibo QIU
Chinese Journal of Emergency Medicine 2016;25(11):1412-1417
Objective To clarify the role of FLT3 signaling-dependent pulmonary conventional dendritic cells (cDCs) in the pathogenesis of lipopolysaccharide (LPS)-induced acute lung injury (ALI),and as well as the modulation effects of cDCs in vivo on the inflammatory responses to acute lung injury.Methods Thirty C57BL/6 male mice were divided into normal control group,LPS group,FLT3L pretreatment group,lestaurtinib,(a high efficient and specific blocker in FLT3 signal pathway) pretreatment group and vehicle (DMSD) control group.FLT3L and lestaurtinib were administrated subcutaneously for 5 days.Murine model of ALI was subsequently established by intra-tracheal application of LPS and lung specimens were harvested 6 h or 24 h later.The accumulation and maturation of pulmonary cDCs were assessed by flow cytometry.IL-6 and TNF-α were quantified to evaluate lung inflammation.Lung injury was estimated by lung wet weight/body weight ratio (LWW/BW) and histopathological assessment.Lung myeloperoxidase (MPO) activity was measured to evaluate neutrophil infiltration.Transcription factors Tbet/GATA-3 mRNA ratio was determined to estimate balance of Th1/Th2 response.IFN-γ and IL-4 were quantified to evaluate Th1-specific and Th2-specific cytokine production respectively.Results The accumulation and maturation of pulmonary cDCs peaked at 6h after LPS challenge.FLT3L pretreatment significantly stimulated the accumulation and maturation of pulmonary cDCs (P < 0.05),leading to markedly deterioration of LWW/BW and lung histopathological changes.Meanwhile lung MPO activity and T-bet/GATA-3 mRNA ratio were elevated (P < 0.05).Furthermore,the production of IL-6,TNF-α and IFN-γwas markedly increased by FLT3L pretreatment (P < 0.05).In contrast,lestaurtinib pretreatment markedly inhibited the accumulation and maturation of pulmonary cDCs (P < 0.05),leading to significant improvement of LWW/BW and lung histopathological changes.Meanwhile lung MPO activity and T-bet/ GATA-3 mRNA ratio were decreased (P < 0.05).Furthermore lestaurtinib efficiently suppressed the production of IL-6,TNF-α and IFN-γ (P < 0.05).Conclusion This study thus demonstrated that FLT3 signaling-dependent pulmonary cDCs could control the initiation of acute lung inflammation response to LPS-induced ALI through the regulation of neutrophil infiltration and balance of Thl/Th2 response.
10.Detection and analysis of T-lymphocyte subpopulation in population exposed to high concentrations of arsenic in drinking water
Yi, GAO ; Guang, HAN ; Jiang, LIANG ; Feng-jie, TIAN ; Qiu-ling, PEI
Chinese Journal of Endemiology 2009;28(4):398-400
Objective To study the change and the significance of T-lymphocyte immune function in peripheral blood in population living in arsenic-contaminated area. Methods Fifty-three cases of patients with arsenism symptoms were selected into experimental group, inhabitants who had no chronic arsenism symptoms into control group in the endemic area of Shuocheng District, Shuozhou City, Shanxi Province in 2006. Vein blood samples were taken and analyzed with SAP assay to measure the percentage of CD3+ ,CD4+ and CD8+ T-cells. Results It was found that the percentage of CD3+, CD4+, and CD4+/CD8+ [(41.89 ± 11.58)%, (25.60 ± 9.05)% and 1.02 ± 0.41] in the experimental group was lower than that in the control group [(68.38 ± 7.23)%, (39.17± 4.28)% ,1.69 ± 0.56, t = 13.61,18.72,14.79, all P < 0.05], while there was no statistical differences of CD8+ [(25.30 ± 6.85)%] compared to the control group[(23.54 ± 8.35)%,t = 3.07,P > 0.05]. The gender-related effect of arsenic on CD4+ and CD8+ was found by multiple linear step regression analysis(t = - 3.05, - 4.30, all P < 0.05). In case group, there were no statistical differences in CD3+, CD4+, CD8+ and CD4+/CD8+[(40.65±10.06)%, (24.48 ± 6.29)%, (24.52 ± 8.16)%,0.98 ± 0.25] between males and females [(43.07±12.96)%, (26.77±3.12)%, (26.50 ±9.32)%, 1.07 ±0.41, t = - 0.76,3.05,0.30,2.10, all P > 0.05]. Conclusions The immune function of T-lymphocytes of patients with chronic arsenism has been suppressed. It is of active significance to detect T-lymphocyte subpopulation in peripheral vein in patients with chronic arsenism aiming at estimating the function of cell immune and providing early diagnosis index.