ABSTRACT
Objective To investigate the protective effect of ligustrazine on the transporting function of hepatocellular mitochondria membrane in the rats with sepsis-induced acute liver injury (SALI). Methods The Sprague-Dawley (SD) rats were randomly divided into sham operation group, SALI group [established by cecal ligation and puncture (CLP)], ligustrazine treatment group (injection of ligustrazine 60 mg/kg through tail vein after CLP) and ligustrazine preventive group (7 days before CLP, ligustrazine was injected daily through tail vein for 60 mg/kg), and there were 12 rats in each group. Abdominal aorta blood and liver were harvested at 10 hours after operation. The content of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and mitochondrial aspartate aminotransferase (m-AST) were determined by enzyme coupling rate method. The content of ATP was detected by colorimetric and chemical fluorescein method. The activity of mitochondrial ATPase was detected by phosphorus quantification. The expressions of mitochondrial membrane aquaporin 8 (AQP8) and carnitine palmitoyl transferase (CPT) were detected by Western Blot. Results Compared with sham operation group, the levels of serum ALT, AST and m-AST were significantly increased in SALI group, ligustrazine treatment group and ligustrazine preventive group, and the content of ATP was reduced, the activity of mitochondrial membrane ATPase, the expressions of AQP8 and CPT-1A were significantly decreased. Compared with SALI group, the levels of serum ALT, AST and m-AST were significantly decreased in ligustrazine treatment and ligustrazine preventive groups [ALT (U/L): 123.8±32.8, 105.0±44.5 vs. 233.0±110.1; AST (U/L):427.0±117.9, 303.9±110.3 vs. 742.6±441.4; m-AST (U/L): 239.6±64.9, 168.2±60.0 vs. 412.8±252.6; all P <0.01], the content of ATP were significantly increased (nmol/mg: 29.5±10.3, 34.6±11.2 vs. 19.3±8.8, both P < 0.01), the activity of ATPase in hepatocellular mitochondrial membrane were significantly increased [Na+-K+-ATPase (U/mg):3.91±0.30, 3.97±0.35 vs. 2.87±0.82; Mg2+-ATPase (U/mg): 3.75±0.38, 3.88±0.35 vs. 2.64±1.06; Ca2+-ATPase (U/mg): 3.15±0.58, 2.98±0.31 vs. 1.75±1.25; Ca2+-Mg2+-ATPase (U/mg): 3.82±0.31, 3.91±0.42 vs. 2.57±1.01, all P < 0.01], the expressions of AQP8 and CPT-1A were significantly increased [percentage increase from sham operation group (100%), AQP8/COX-Ⅳ: (79.12±7.79)%, (88.40±9.22)% vs. (62.08±11.91)%; CPT-1A/COX-Ⅳ:(87.92±10.06)%, (84.91±17.48)% vs. (72.11±7.82)%, all P < 0.01]. The levels of serum AST and m-AST in ligustrazine preventive group were significant lower than those in ligustrazine treatment group [AST (U/L): 303.9± 110.3 vs. 427.0±117.9; m-AST (U/L): 168.2±60.0 vs. 239.6±64.9, both P < 0.05]. There was no significant difference in the expression of CPT-2 in mitochondrial membrane between the four groups. Conclusions Ligustrazine could play a protective role on the mitochondrial membrane function of transporting water, ion and fat in the rats with SALI. The preventive function of ligustrazine is better than the treatment effect of the rats with sepsis.
ABSTRACT
Objective To observe the effects of Angelica Sinensis Radix on the expression of aquaporin 8 (AQP8) and the levels of AC-cAMP-PKA in colon of blood-deficiency constipation in mice; To identify mechanism of Angelica Sinensis Radix for loosening the bowels to relieve constipation. Methods Sixty Kunming mice were randomly divided into control group, model group, positive medicine group, and Angelica Sinensis Radix high-, medium-, and low-dose groups, with ten mice in each group. Diphenoxylate, acetylphenlyhydrazine and cyclophosphamide were used to establish blood-deficiency constipation mice models. From the 14thday of the experiment, Angelica Sinensis Radix high-, medium-, and low-dose groups were given 16.7, 8.8, and 4.2 g/kg Angelica Sinensis Radix Decoction for gavage. Positive medicine group was given 5.0 g/kg Changtongshu Granules Liquid for gavage. Control group and model group were given equal volume of saline for gavage. The symptoms of blood deficiency constipation were observed and defecation time. Immunohistochemistry, Western blot and qRT-PCR were used to detect the expression of AQP8 protein and mRNA in colon, and the expression of AC-cAMP-PKA in colon was detected by ELISA. Results The mice in the model group developed blood deficiency constipation syndrome; the defecation time was significantly prolonged (P＜0.01); the expression level of colonic AQP8 protein and mRNA, AC, cAMP and PKA significantly increased (P＜0.05, P＜0.01). Compared with model group, the defecation time was significantly shortened in Angelica Sinensis Radix high-, medium-, and low-dose groups; the expression of AQP8 protein and mRNA and the levels of AC, cAMP and PKA were significantly decreased (P＜0.05, P＜0.01). Conclusion The treatment of Angelica Sinensis Radix for blood-deficiency constipation may be related to adjusting the AC-cAMP-PKA signaling pathways and reducing the expression of AQP8 protein and mRNA.
ABSTRACT
OBJECTIVE: To observe the effects of Modified Rhizoma Alismatis decoction on the expression of aquaporin 8 (AQP8) in liver tissue of hyperlipemia model rats, and to investigate the mechanism of preventing and treating hyperlipemia.METHODS: Total of 60 rats were randomly divided into blank control group (distilled water), model group, positive control group (simvastatin 1. 89 mg/kg) and modified Rhizoma Alismatis decoction high-dose, medium-dose and low-dose groups (29. 56, 14. 78, 7. 39 g/kg, calculated by crude drug), with 10 rats in each group. Those groups were given high-fat diet to induce hyperlipemia model and given relevant medicine intragastrically once a day for consecutive 5 weeks except that blank control group was given normal diet. After administration, the serum contents of TG, TC, HDL-C and LDL-C in rats were detected, and the pathomorphology changes of liver tissue were observed; the mRNA and protein expression of AQP8 in liver tissue were detected. RESULTS: Compared blank control group, the serum contents of TG, TC and LDL-C in model group were increased significantly (P<0. 05 or P<0. 01),while the serum content of HDL-C was decreased significantly (P<0. 01); pathological changes were found in liver tissue, such as irregular cell arrangement and hepatic sinusoidal hyperemia and edema; mRNA and protein expression of AQP8 in liver tissue were increased significantly (P<0. 01). Compared with model group, above indexes of treatment groups were improved significantly (P<0. 05 or P<0. 01); the structure of liver tissue tended to be normal and the fatty degeneration was obviously alleviated. CONCLUSIONS: Modified Rhizoma Alismatis decoction can regulate the mRNA and protein expression of AQP8 in liver tissue so as to play the effects on the prevention and treatment of hyperlipidemia.
ABSTRACT
Objective To observe the effect of Yang-warming and Qi-tonifying Recipe (YQP)on aquaporin 3(AQP3) and AQP8 in rats with slow transit constipation,and to explore its therapeutic mechanism. Methods Forty rats were randomly divided into modeling group(N=30)and normal control group(N = 10). After successful modeling by gastric gavage of loperamide,the modeled rats were randomly divided into model group,YQP group and Mosapride group,10 rats in each group,and were separately treated with corresponding medicine for 2 weeks. After treatment, the colonic transit function was measured by carbon propelling test. The protein levels of AQP3 and AQP8 were detected by immunohistochemistry and their mRNA expression levels were detected with real-time fluorescence polymerase chain reaction (qPCR). Results Compared with the normal control group,the propelling rate of carbon particle in the model group was decreased,and the protein and mRNA expression levels of AQP3 and AQP8 were significantly increased(P<0.05 or P<0.01). Compared with the model group,the propelling rate of carbon particle of YQP group and mosapride group was significantly increased, and the protein and mRNA expression levels of AQP3 and AQP8 were significantly decreased (P<0.05),but there was no significant difference between YQP group and mosapride group (P >0.05). Conclusion YQP had therapeutic effects on loperamide-induced constipation through decreasing the expression of AQP3 and AQP8 in the intestine,reducing the reabsorption of intestinal fluid, and increasing the fecal water content.
ABSTRACT
Objective To detect the expression level of aquaporin 8 (AQP8) in patients with functional constipation(FC) or constipated irritable bowel syndrome (IBS-C),and the correlation between the expression of AQP8 and clinical features.Methods From March to December 2014,a total of 16 patients with IBS-C and 19 patients with FC met Rome Ⅲ criteria were collected,and nine healthy individuals were assigned to control group at the same period.The ascending and decending colonic tissues mucosa of FC,IBS-C and control group were taken under endoscope.The expression of AQP8 at mRNA and protein level was detected by real-time polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC).The differences in AQP8 mRNA expression and AQP8 relative area were analyzed by Kruskal-Wallis test among groups,and Pearson correlation coefficient was performed for correlation analysis between the expression and clinical features.Results The relative expressions of AQP8 mRNA of ascending colon and descending colon of FC patients (1.38,0.61 to 4.09;2.65,0.82 to 7.52) and IBS-C patients (2.23,0.82 to 4.67;1.35,0.51 to 2.03) were higher than those of control group (0.56,0.19 to 0.97;0.38,0.21 to 1.19),and the differences were statistically significant (ZFc =-2.435,-3.149,ZIBS-C =-2.690,-2.152;all P＜0.05).AQP8 mRNA expression of descending colon in patients with FC was higher than that of patients with IBS-C,and the difference was statistically significant (Z =-2.003,P =0.045).The expression of AQP8 in patients with FC and IBS-C was positively correlated with disease course (ascending colon r=0.57 and 0.53;descending colon r=0.49 and 0.54,all P＜0.05),and was negatively correlated with frequency of defecation (ascending colon r=-0.82 and-0.61;descending colon r=-0.49 and-0.53,all P＜0.05).There was no correlation between the expression of AQP8 and age,gender,onset age,presence of abdominal symptoms of the patients (all P＞ 0.05).Most of AQP8 of FC group was expressed in cytoplasm of colonic mucosa epithelial cells,while that of IBS-C group and control group was mostly expressed at apical membrane and basal membrane of epithelial cells.The results of semi-quantification demonstrated that AQP8 relative area of descending colon of FC and IBS-C group increased compared with that of control group (3.42％ (1.24％ to 5.61％),2.45％(1.72％ to 4.27％) vs 1.18％ (0.35％ to 2.81％);Z=-2.534,-2.151,both P＜0.05).Meanwhile,AQP8 relative area of ascending colon of FC group increased compared with that of control group (2.46％(1.48％ to 4.18％) vs 1.14％(1.29％ to 2.15％) Z=-2.041,P＜0.05).Conclusion There are differences in AQP8 expression quantity and location in cells of descending colon between patients with FC and patients with IBS-C,which is a way for differentiation these two diseases.
ABSTRACT
OBJECTIVE: This study was to determine whether aquaporin-8, which plays a role as a transcellular water channel, is expressed in human placenta, and to compare the degree of its expression between preeclamptic women and normal pregnant women. METHODS: Placentas were obtained from severely preeclamptic women and normal pregnant women who were delivered babies by cesarean section before the onset of labor in the Chungbuk National University Hospital. In situ hybridization with aquaporin-8 cRNA probe was performed using paraffin-embedded tissue section. Signal of aquaporin-8 expression was observed with light microscope. RESULTS: In situ hybridization demonstrated strong expression of aquaporin-8 mRNA in the placentas of both preeclamptic women and normal pregnant women. The degree of expression was the same in both group. CONCLUSION: Aquaporin-8 in human placenta may not be related to the pathogenesis of preeclampsia.
Subject(s)
Female , Humans , Pregnancy , Cesarean Section , In Situ Hybridization , Placenta , Pre-Eclampsia , Pregnant Women , RNA, Complementary , RNA, Messenger , WaterABSTRACT
Objective:To determine the expression of aquaporin 8 and its localization in human WISH cells and to explore the molecular and cellular mechanisms for water absorption across the amniotic membranes.Methods:Human amnion-derive WISH cells were cultured.Western analysis was used to quantify AQP8 expression level.Reverse transcripton-polymerase chain reaction(RT-PCR)was used to quantify AQP8 mRNA expression levels.Immunofluorescence was used to determine localization of AQP8 in WISH cells.Results:AQP8 mRNA protein was found in human WISH cells.By western-blot,the AQP8 proteim was detected at 34ku in human WISH cells.AQP8 labeling was observed in intracellular vesicular structures throughout the cytosol and plasma membrane.Conclusion:The study demonstrates the expression of aquaporin 8 in human WISH cells.These results suggest that aquaporin 8 may be a channel that mediates amniotic fluid resorption by way of intramembranous pathway.