ABSTRACT
OBJECTIVE: To investigate the role of N-methyl-D-aspartate receptor(NMDAR)/cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA) signaling pathways in regulating 2-chloroethanol-induced aquaporin-4(AQP4) expression in astrocytes(AS). METHODS: i) AS in logarithmic growth phase were treated with 2-chloroethanol at the doses of 0.0, 7.5, 15.0 and 30.0 mmol/L for 12 hours, and the cells were collected for detection. ii) The AS in logarithmic growth phase were divided into blank control group, inhibitor control group, 2-chloroethanol group, and inhibitor intervention group. The inhibitor included dizocilpine(MK-801) and N-(2-[p-bromocinnamylamino-]ethyl)-5-isoquinolinesulfonamide(H89). The blank control group did not receive any treatment. The inhibitor control group was treated with a concentration of 10.0 μmmol/L MK-801 or 15.0 μmmol/L H89. The MK-801 intervention group was pretreated with MK-801 at a concentration of 10.0 μmmol/L for 30 minutes. The H89 intervention group was pretreated with H89 at a concentration of 15.0 μmmol/L for 1 hour. After the intervention, the AS in 2-chloroethanol group and MK-801, H89 intervention group were stimulated with 2-chloroethanol at a dose of 30.0 mmol/L for 12 hours. iii) The AS in each group were collected and used for Western blotting and real-time fluorescence quantitative polymerase chain reaction analysis to detect the protein and mRNA expression of AQP4, NMDAR receptor main subunit(NR1), NMDAR receptor 2 B subunit(NR2 B) and calmodulin dependent protein kinaseⅡ(CaMKⅡ). The Western blotting was adopted to detect the expression of phosphorylase-CaMKⅡ(p-CaMKⅡ) and PKA. Colorimetric method was used to detect the concentration of calcium(Ca~(2+)) in AS. The enzyme-linked adsorption test was used to measure adenylate cyclase(AC) activity and cAMP levels. RESULTS: i) The relative expression of protein and mRNA of AQP4, NR1 and NR2 B, PKA at protein level and CaMKⅡ at mRNA level, and the ratio of p-CaMKⅡ/CaMKⅡ protein, the concentration of Ca~(2+), AC activity and cAMP level in 30.0 mmol/L group were higher then those of 0.0 mmol/L group in AS(P<0.05). The relative protein expression of PKA and the concentration of Ca~(2+) increased with the increase of 2-chloroethanol(P<0.05). ii) The relative protein expression of AQP4 and the concentration of Ca~(2+) in the 2-chloroethanol group were higher than that of the blank control group and MK-801 control group(P<0.05). The relative protein expression of AQP4 and the concentration of Ca~(2+) in MK-801 intervention group were lower than that in 2-chloroethanol group(P<0.05). The relative protein expression of AQP4 and PKA in 2-chloroethanol group were higher than that of the blank control group and H89 control group(P<0.05). The relative protein expression of AQP4 and PKA in H89 intervention group was lower than that in 2-chloroethanol group(P<0.05). CONCLUSION: The 2-chloroethanol timulation induces the expression of AQP4 by activating NMDAR/cAMP/PKA signaling pathway in AS.
ABSTRACT
Objective To explore the effect of the modified Sheng operation for adult proctoptosis Methods The modified Sheng operation was performed on 13 adult patients with whole proctoptosis. The average 3.5 years postoperative follow-up was done. Results 12 case were cured(12/13). one case was obvionsly improved (1/13). Conclusions The modified Sheng operation is a satisfactory procedure in treating adult proctoptosis. It does not only treat rectum and tedious sigmoid , but also correct the parenteral factor ,such as pelvic floor hernia , uterine compression etc.