Objective To investigate the changes of renal medulla aquaporin 2 expression and morphological changes of epithelia of collecting tube after bile duct recanalizaiton operation. Methods Thirty rats were divided into two groups randomly. Common bile duct ligation was performed on 20 experimental rats with silicon tubes 2 mm in extre-diameter, and sham operation on the other 10 rats. Seven days later, bile duct recanalizaiton was performed on obstructive jaundice group and sham operation on contrast group. Experimental rats were divided into two subgroups randomly. Half of them were killed immediately and the others would be killed 24 hours later. Serum of each rat was collected to detect hepatic function and renal function. Renal medulla was fixed for microscopic examination and was kept in the -80 ℃ refrigerator for aquaporin 2 expression measurement by Western blot technique. Results All of the animals accomplished the experiment smoothly. Golden ascites were found in the rats of obstructive jaundice group. Twenty-four hours after recanalization, serum bilirubin levels decreased 〔(45.95±8.39) μmol/L〕, P＜0.01, and there was no significant change in blood urine and creatine level. Compared with sham operation group (21 966.20±1 544.70), expression of aquaporin 2 decreased significantly after common bile duct ligation in obstructive jaundice group (15 665.30±1 181.85), P＜0.01. After recanalizaion, the expression of aquaporin 2 in obstructive jaundice group increased (19 490.80±4 239.32), P＜0.01. Conclusion Common bile duct obstruction would lead to epithelium injury of renal collecting tube, and down regulate the aquaporin 2 expression.
ObjectiveTo observe the effects of aquaporin 1 (AQP1) on the proliferation and migration of endothelial progenitor-endothelial progenitor cells (EPC).MethodsBone marrow cells of AQP1 wild-type (WT) (n=6) and knockout-type (KO) mice (n=6) were isolated and differentiated into EPC in vitro. Immunofluorescence was used to detect cell surface antigens to identify EPC. Live cell kinetic imaging and quantification technology, transwell migration assays, as well as scratch test were used to compare the function of EPC between AQP1 WT and KO mice.ResultsEPC culture showed that cells were initially suspended and gradually adhered to typical mesenchymal stem cells within 7 days. After cultured on special medium for endothelial cells they were adhered and differentiated, and fusiform or polygonal, paving stone-like EPC were observed around 14 days. When cultured by special medium of EPC, CD133 and CD31 were positively detected after 7 days, and CD34 and Flk-1 were positively detected after 14 days. Positive expression of AQP1 was only detected in EPC of AQP1 WT mice. Functional studies of EPC revealed there was no significant difference in the proliferation of EPC between AQP1 WT and KO group mice. Transwell assay showed that EPC migration ability of AQP1 KO mice was significantly weaker than that of WT mice. The scratch healing ability of EPC in AQP1 KO mice was significantly lower than that of WT mice.ConclusionsEPC initially shows the characteristics of stem cells and with the prolongation of culture time, EPC gradually shows the characteristics of endothelial cells. AQP1 affects the EPC migration rather than proliferation.
Objective To observe the correlation of serum aquaporin 4 (AQP4) antibodies and condition and visual prognosis in patients with severe neuromyelitis optica spectral disorders (NMOSD). Methods Fifty NMOSD patients with visual acuity of 20/200 or worse in at least one eye were enrolled in this retrospective analysis. There were 12 males and 38 females. The age ranged from 17 to 65 years, with the mean of (39.86±2.02) years. The patients were divided into two groups according to the serum AQP4-IgG status. The ophthalmologic examination, serum anti-nuclear antibodies (ANA), myelin oligodendrocyte glycoprotein (MOG) antibody detection and vision prognosis were compared and analyzed. Glucocorticoid therapy was delivered to 46 patients who were within 1 month of onset. The visual acuity of the patients after treatment was divided into complete recovery, partial recovery, stabilization and reduction, and the visual acuity of the two groups were analyzed. Results Among 50 patients, there were 30 (60%) seropositive patients (positive group), 20 (40%) seronegative patients (negative group). The positive group had significantly higher ratio of female to male (P=0.004), and more binocular optic neuritis (ON) (P=0.010) compared with the negative group. More recurrence ON were also found in the positive group, but without statistic difference between two groups (P=0.167). There was no difference of age, course, and vision damage degrees and abnormal orbital MRI scanning between two groups (P＞0.05). Among 24 patients who underwent serum ANA detection in the positive group, 8 patients were positive. All of 18 patients who underwent serum ANA detection in the negative group were negative. The difference of the ratio of serum ANA positive patients between two groups was significant (P=0.030). Serum MOG antibody detection in the positive group was negative (0/10). Sixteen patients who underwent MOG antibody detection in negative group, 4 patients were positive. After treatment, there were 23.3%, 23.3%, 53.3% patients with vision of complete recovery, partial recovery and reduction in the positive group; 25.0%, 30.0%, 25.0% patients with vision of complete recovery, partial recovery and reduction in the negative group, respectively. There was no difference in proportion of vision with complete recovery and partial recovery between two groups (P=0.163, 0.607), but significant difference was observed in proportion of vision with stabilization and reduction between two groups (P=0.021, 0.048). Conclusions The positive serum AQP4 antibody is common in patients with severe NMOSD. The patients with AQP4 antibody in the serum are more likely combined with immunological serological markers and poor vision prognosis.
ObjectiveTo observe the ocular manifestations and the titer of aquaporin 4 antibody (AQP-4) in NMO patients, and to evaluate the BCVA prognosis in patients with different titers of AQP-4Ab.MethodsA retrospective case study. From September 2009 to March 2014, 132 NMO patients diagnosed in Department of Neurology and Ophthalmology in Huashan Hospital of Fudan University were included in the study. Among the patients, 74 patients (56.06%) were involved in optic nerve for the first time, among which 63 patients (47.72%) were involved in optic nerve alone, and 11 patients (8.33%) were involved in optic nerve and spinal cord at the same time. The recurrence rate was 62.88% (twice or more). All patients underwent BCVA, slit lamp microscope, fundus examination, thyroid function, sex hormones, and serum AQP-4Ab detection. BCVA was recorded at admission and before discharge from hospital, and worse BCVA was recorded in binocular patients. The BCVA of patients with different titers of AQP-4Ab were analyzed comparatively.ResultsAmong the 74 patients with optic nerve involved in the first onset, 50 patients with BCVA＜0.1 at the initial diagnosis (67.57%); AQP-4Ab positive was found in 56 patients, which including 13, 9 and 34 patients of AQP-4Ab titer 5 - 60, 61 - 100 and ＞100 RSRU/ml. After 2 weeks of treatment, BCVA improved in 40 patients (71.42%), including 11 (84.62%), 6 (66.67%) and 23 (67.64%) of AQP-4Ab titer 5 - 60, 61 - 100 and ＞ 100 RSRU/ml. Among 132 patients, 98 patients (74.24%) were AQP-4Ab positive. There were 73 patients (55.30%) with abnormal immune rheumatoid index.ConclusionsThe optic nerve is involved in 56.06% patients with NMO for the first time, and 67.57% of the patients had poor vision with BCVA＜0.1. BCVA prognosis is better in patients with serum AQP-4Ab titer of 5 - 60 RSRU/ml.
【摘要】 目的 探讨腹水引起的腹内高压对肝硬化小鼠肺组织水通道蛋白1（AQP1）和水通道蛋白5（AQP5）表达的影响。 方法 雄性美国癌症研究所（Institudo of Cancer Reseach，ICR）小鼠50只，随机取10只作正常对照组（腹压0 cm H2O，1 cm H2O=0.098 kPa），其余40只用四氯化碳建立肝硬化小鼠模型，并随机分为4组:肝硬化（腹压0 cm H2O）组、肝硬化（腹压5 cm H2O）组、肝硬化（腹压10 cm H2O）组、肝硬化（腹压20 cm H2O）组，通过腹腔注射不同量的白蛋白生理盐水形成不同的腹压，并维持腹压24 h后取肺组织行病理、免疫组织化学、肺湿/干比值及实时荧光定量PCR检测AQP1和AQP5 mRNA表达量。 结果 与正常对照小鼠相比，肝硬化小鼠肺AQP5、AQP1表达明显下降（Plt;0.05）；肝硬化小鼠随着腹内压的升高，肺湿/干比值升高，AQP5、AQP1表达相应增加（Plt;0.05）。 结论 肝硬化可以影响肺AQP1、AQP5的表达；肝硬化小鼠随着腹内压的升高，AQP1、AQP5表达相应增加，并与肺水肿的严重程度密切相关。【Abstract】 Objective To investigate the role of intra-abdominal hypertension caused by ascites on the expression of Aquaporin (AQP) 1 and AQP 5 in the lung of cirrhotic mice. Methods We randomly chose 10 from 50 male Institude of Cancer Research (ICR) mice to form the control group [intra-abdominal pressure (IAP)=0 cm H2O, 1 cm H2O=0.098 kPa]. The model of cirrhosis were prepared by subcutaneous injection of carbon tetrachloride for the rest 40 mice which were then randomly divided into 4 groups: cirrhosis (IAP=0 cm H2O) group, cirrhosis (IAP=5 cm H2O) group, cirrhosis (IAP=10 cm H2O) group, and cirrhosis (IAP=20 cm H2O) group. Saline with different volume of albumin was injected into the peritoneum of each mouse in order to form different IAP. After 24 hours, analysis of pathology, immunochemistry and wet/dry ratio was done for the lungs of these mice; and the expression of AQP1 and AQP5 at the protein and mRNA levels were analyzed by IHC and qRT-PCR. Results Compared with the normal mice, the expression of AQP1 and AQP5 in lungs of cirrhotic mice were significantly lower (Plt;0.05). Both the lung wet/dry ratio and the expression of AQP1 and AQP5 raised with the increase of IAP. Conclusion Cirrhosis can affect the expression of AQP1 and AQP5 in lungs. The expression of AQP5 and AQP1 in lungs of cirrhotic mice increases with the increase of IAP, which is also closely correlated with the severity of pulmonary edema.
ObjectiveTo analyze the clinical features and prognosis of adult optic neuritis patients with positive serum myelin oligodendrocyte glycoprotein antibody (MOG-ON) or aquaporin 4 antibody (AQP4-ON).MethodsA retrospective study. From December 2015 to February 2018, in the Beijing Chaoyang Hospital of Capital Medical University and Chinese PLA General Hospital, 162 eyes of 132 patients with positive serum MOG antibody and AQP4 were included in the study. There were 42 MOG-ON patients (49 eyes, 31.8%), 90 AQP4-ON patients (113 eyes, 68.2%). The clinical features of optic neuritis (annual recurrence frequency, incidence of optic disc edema), brain and optic nerve enhanced MRI, serum autoimmune antibodies and cerebrospinal fluid test results were compared between MOG-ON and AQP4-ON patients. All patients were treated with intravenous methylprednisolone sodium succinate in the acute phase and then switched to oral prednisone acetate tablets. The average follow-up time was 15 months. The glucocorticoid dependence, visual prognosis, spinal cord symptoms, and myelitis at the last follow-up were comparatively analyzed between MOG-ON and AQP4-ON patients. The comparison of the count data was performed by χ2 test, and the measurement data were compared by t test.ResultsCompared with AQP4-ON patients, MOG-ON patients had higher annual recurrence frequency (t=3.760, P=0.005), higher incidence of optic disc edema (χ2=14.777, P＜0.001), higher incidence of hormone dependence (χ2=25.496, P＜0.001), and better visual prognosis (χ2=28.759, P＜0.001). MOG-ON patients were more likely to involve the optic nerve, AQP4-ON patients were more likely to involve the optic chiasm and the optic tract. There was a significant difference in the location of lesions between MOG-ON and AQP4-ON patients (χ2= 5.447, P= 0.015). The proportion of AQP4-ON patients with autoimmune antibodies was significantly higher than that of MOG-ON patients (χ2 = 20.453, P＜0.001). The results of cerebrospinal fluid test showed that the white blood cell count of patients with MOG-ON and AQP4-ON were within the normal range, but the IgG level of AQP4-ON patients was significantly higher than that of MOG-ON patients (t=8.669, P＜0.001). At the last follow-up, there were 7 and 29 patients of myelitis in MOG-ON and AQP4-ON patients respectively (χ2=3.494, P=0.046).ConclusionsThe clinical characteristics of MOG-ON were different from AQP4-ON. The incidence of optic disc edema and recurrence rate were higher, but the proportion of autoimmune antibodies was lower. MOG-ON was more likely to show hormone dependence, but the visual prognosis was better. AQP4-ON was easily involved in optic chiasm and optic tract, and the incidence of myelitis was higher.
Objective To investigate the effects of extracts of Pinellia ( EP) on a rat model of airway mucus hypersecretion induced by LPS. Methods Thirty Wisatr rats were randomly divided into 5 groups, ie. a blank group, a model group, and three EP groups treated with different doses of EP. There were 6 rats in each group. Airway mucus hypersecretion model was established by intratracheally instillation of LPS in the model group and three EP groups. The rats in three EP groups were orally administered with EP at dosages of 10 g/kg, 30 g/kg and 60 g/kg respectively for 4 days. The expression of Mucin 5AC ( MUC5AC) protein in airway was assayed by immunohistochemistry. The mRNA expressions of MUC5AC and Aquaporin-5( AQP-5) in lung tissue were detected by RT-PCR. ELISA technique was performed to detect TNF-αin bronchoalveolar lavage fluid( BALF) . Results LPS significantly stimulated the mRNA and protein expression of MUC5AC in lung and TNF-αlevel in BALF, and inhibited the expression of AQP-5 mRNA in lung. The EP at dosages of 10 g / kg and 30 g/ kg had little effect on mucus hypersecretion. While 60 g/kg of EP could significantly inhibited the expression of MUC5AC, and decreased the release of TNF-α in BALF. The AQP-5 mRNA was also up-regulated by 60 g /kg of EP. The expression of MUC5AC mRNA was positively correlated with level of TNF-α( r = 0. 948, P lt;0. 05) ; AQP-5 mRNA was negatively correlated with MUC5AC mRNA and TNF-α( r = - 0. 955, P lt; 0. 05; r = - 0. 909, P lt; 0. 05) . Conclusion EP ( 60 g/ kg) can significantly attenuated airway mucus hypersecretion in rats.
ObjectiveTo explore the value of Aquaporin-3 (AQP-3) on the detection of early renal function damage by investigating the expressions of renal AQP-3 mRNA and protein of rats with obstructive jaundice (OJ). MethodsForty mature male Wistar rats were divided into two groups randomly: experimental group (n=20) in which the model of OJ rats was established, and control group (n=20, sham operation group). The levels of serum total bilirubin (TBIL), direct bilirubin (DBIL), creatinine (Cr)， and blood urea nitrogen (BUN) were detected by fullautomatic biochemical analyzer on 7 d and 14 d after operation. The expressions of renal AQP-3 mRNA and protein of rats were detected by RT-PCR and Western blotting, respectively. ResultsThe levels of serum TBIL and DBIL were significantly higher on 14 d than those on 7 d after operation in experimental group (P=0.000), which were significantly higher than those at corresponding time point in control group (P=0.000), while the difference within control group was not significant (P=0.154). Thus, the OJ models of rats were established successfully. The difference of serum Cr levels of rats between inter-and intragroup were not significant (Pgt;0.05). Serum BUN level on 14 d after operation in experimental group was significantly higher than those on 7 d after operation in experimental group and on 14 d after operation in control group (P=0.001), although serum Cr levels were not different between 7 d and 14 d after operation in control group (P=0.288). The expressions of AQP-3 protein of rats on 7 d and 14 d after operation in experimental group were significantly lower than those at corresponding time point in control group (P=0.033, P=0.000), meanwhile on 14 d after operation in experimental group was significantly lower than those on 7 d after operation in experimental group (P=0.000). The expressions of AQP-3 mRNA of rats on 7 d and 14 d after operation in experimental group were significantly higher than those at corresponding time point in control group (P=0.000), but the difference at different time point in two groups was not significant (P=0.139, P=0.059). ConclusionsThe changes of renal AQP-3 protein and mRNA expressions are prior to the changes of serum Cr and BUN levels of rats suffered from OJ complicated renal function damage, which are promised to improve the early diagnosis rate of renal function damage in rats with OJ.
Objective To investgate the expression of p38 mitogen-activated protein kinase （p38MAPK） in lung tissue of rats with severe acute pancreatitis （SAP）， and to explore the relationship between p38MAPK and pulmonary capillary barrier injury. Methods Forty male and healthy Sprague-Dawley （SD） rats were randomly （random number method） divided into sham operation （SO） group and SAP group， then rats of SAP group were sub-divided into 3， 6， 12， and 24 h group， each group enrolled 8 rats， respectively. SAP model rats were established by injecting 5% sodium taurocholate solution retrograde into the biliopancreatic duct. ELISA method was used to test the serum tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β）， and pathological changes in lung and pancreas tissues were observed by HE staining. Immunohischemistry method was used to detect phosphorylated p38 （p-p38） protein and aquaporin 1 （AQP1） protein of lung tissues. The expression level of AQP1 mRNA was measured by quantitative real-time PCR. Results Hyperemia， edema， and inflammatory cell infiltration were observed in lung tissues， abundance of necrosis， part gland structure fuzzy or even disappear were observed in pancreas tissues of all 4 time point groups. Compared with SO group， levels of serum TNF-α and IL-1β were significantly higher in 4 time point groups （P＜0.05）. Lower expression level of p-p38 protein was detected in lung tissues of SO group， while in the early stage of SAP （SAP 3 h group）， the expression level of p-p38 protein significantly increased， which peaked in 6 h group and was still higher than SO group in 24 h group （P＜0.05）. Compared with SO group， the expression levels of AQP1 mRNA and protein were significantly lower in all 4 time point groups （P＜0.05）， which had negative correlation with the levels of serum TNF-α，IL-1β， and the expression level of p-p38 protein （r=-0.87， P＜0.05；r=-0.88， P＜0.05；r=-0.78， P＜0.05）. Conclusion The decrease of AQP1 protein in lung tissue is one of the vital causes for pulmonary capillary barrier injury in SAP， which probably works by the activation of p38MAPK and the excessive release of inflammatory cytokines.