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Objective@#To study the effects of estrogen on bicarbonate secretion of duodenal mucosal, and to observe estrogen receptor (ER) subtypes of estrogen.@*Methods@#Sixteen 4-week-old male C57 mice were divided into control group and estrogen group, with eight mice in each group. The mice serum level of estrogen was detected by chemiluminescence. The expression of cystic fibrosis transmembrane conductance regulator (CFTR), solute carrier family 26 (SLC26) A3 and SLC26A6 in the duodenum tissues were determined by real-time polymerase chain reaction (RT-PCR). After SCBN cells treated with estrogen, ERα and ERβ blocking agent, and transfected with silenced ERα and ERβ for 24 and 48 hours, the expression levels of CFTR, SLC26A3 and SLC26A6 mRNA in cells were detected by RT-PCR. The effects of estrogen before and after silenced ERα and ERβ on bicarbonate secretion of SCBN cells were observed by high-speed ion imaging system. T test and rank sum test were used for statistical analysis.@*Results@#Compared with that of control group, the serum estrogen level of estrogen group was significantly high ((4 874±942) pmol/L vs. (657±187) pmol/L, t=-11.579, P<0.01). The expression levels of CFTR, SLC26A3 and SLC26A6 mRNAs in duodenum tissues of estrogen group were higher than those of control group (0.856±0.302 vs. 0.452±0.246, 2.910±1.680 vs. 1.120±0.540, 1.272±0.667 vs. 0.319±0.114), and the differences were statistically significant (t=-2.317, -2.483 and -3.721, all P<0.05). Compared with those treated with estrogen for 24 and 48 hours, the levels of CFTR mRNA and SLC26A6 mRNA were lower after the ERβ blocking agent were added into estrogen for 24 and 48 hours (CFTR mRNA: 0.171±0.059 vs. 0.522±0.260 and 0.111±0.014 vs. 0.578±0.297; SLC26A6 mRNA: 0.486±0.289 vs. 1.118±0.571 and 0.492±0.231 vs. 1.551±0.605), and the differences were statistically significant (tCFTR mRNA=2.974 and 2.655, tSLC26A6 mRNA=2.393 and 3.272; all P<0.05). Compared with those of silenced ERα group, the levels of CFTR mRNA, SLC26A3 mRNA and SLC26A6 mRNA were higher after ERα silenced and then added estrogen for 24 and 48 hours (24 h: 5.073±2.270 vs. 1.185±0.494, 1.796±1.168 vs. 0.468±0.108 and 3.085±1.357 vs. 0.706±0.347; 48 h: 5.025±1.998 vs. 1.185±0.494, 1.557±0.653 vs. 0.468±0.108 and 3.290±1.750 vs. 0.706±0.347), and the differences were statistically significant (t24 h=-4.122, -2.773 and -4.162, t48 h=-4.604, -4.034 and -3.250; all P<0.05). Compared with that of silenced ERα group, the bicarbonate secretion increased after ERα silenced and then added estrogen for 24 and 48 hours (0.72±0.17 and 1.15±0.25 vs. 0.35±0.17), and pH also elevated, and the differences were statistically significant (t=-6.516 and -12.387, both P<0.01).@*Conclusion@#Estrogen mainly up-regulates the expression of bicarbonate transporter protein in duodenal mucosal epithelial cells through ERβ, and promotes the bicarbonate secretion of duodenal mucosa.
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Objective To study the effects of estrogen on bicarbonate secretion of duodenal mucosal , and to observe estrogen receptor (ER) subtypes of estrogen.Methods Sixteen 4-week-old male C57 mice were divided into control group and estrogen group , with eight mice in each group .The mice serum level of estrogen was detected by chemiluminescence .The expression of cystic fibrosis transmembrane conductance regulator (CFTR), solute carrier family 26 (SLC26) A3 and SLC26A6 in the duodenum tissues were determined by real-time polymerase chain reaction ( RT-PCR).After SCBN cells treated with estrogen , ERαand ERβblocking agent, and transfected with silenced ER αand ERβfor 24 and 48 hours, the expression levels of CFTR, SLC26A3 and SLC26A6 mRNA in cells were detected by RT-PCR.The effects of estrogen before and after silenced ER αand ERβon bicarbonate secretion of SCBN cells were observed by high-speed ion imaging system.T test and rank sum test were used for statistical analysis .Results Compared with that of control group , the serum estrogen level of estrogen group was significantly high ((4 874 ±942) pmol/L vs.(657 ±187) pmol/L,t＝-11.579, P?0.01). The expression levels of CFTR, SLC26A3 and SLC26A6 mRNAs in duodenum tissues of estrogen group were higher than those of control group (0.856 ±0.302 vs.0.452 ±0.246, 2.910 ±1.680 vs.1.120 ±0.540, 1.272 ± 0.667 vs.0.319 ±0.114), and the differences were statistically significant ( t ＝-2.317,-2.483 and-3.721, all P?0.05).Compared with those treated with estrogen for 24 and 48 hours, the levels of CFTR mRNA and SLC26A6 mRNA were lower after the ERβblocking agent were added into estrogen for 24 and 48 hours (CFTR mRNA: 0.171 ±0.059 vs.0.522 ±0.260 and 0.111 ±0.014 vs.0.578 ±0.297; SLC26A6 mRNA:0.486 ±0.289 vs.1.118 ±0.571 and 0.492 ±0.231 vs.1.551 ±0.605), and the differences were statistically significant (tCFTR mRNA＝2.974 and 2.655, tSLC26A6 mRNA＝2.393 and 3.272; all P?0.05).Compared with those of silenced ERαgroup, the levels of CFTR mRNA, SLC26A3 mRNA and SLC26A6 mRNA were higher after ERα silenced and then added estrogen for 24 and 48 hours (24 h: 5.073 ±2.270 vs.1.185 ±0.494, 1.796 ±1.168 vs.0.468 ±0.108 and 3.085 ±1.357 vs.0.706 ±0.347; 48 h: 5.025 ±1.998 vs.1.185 ±0.494, 1.557 ± 0.653 vs.0.468 ±0.108 and 3.290 ±1.750 vs.0.706 ±0.347), and the differences were statistically significant (t24 h＝-4.122,-2.773 and -4.162, t48 h ＝-4.604,-4.034 and -3.250; all P?0.05).Compared with that of silenced ERαgroup, the bicarbonate secretion increased after ER αsilenced and then added estrogen for 24 and 48 hours (0.72 ±0.17 and 1.15 ±0.25 vs.0.35 ±0.17), and pH also elevated, and the differences were statistically significant (t＝-6.516 and -12.387, both P?0.01).Conclusion Estrogen mainly up-regulates the expression of bicarbonate transporter protein in duodenal mucosal epithelial cells through ER β, and promotes the bicarbonate secretion of duodenal mucosa .
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Objective To investigate the effect and potential mechanism of autophagy inhibitor chloroquine on the calcium oxalate crystals formation in rats.Methods From September 2016 to October 2016,Thirty healthy male SD rats were randomly divided into 3 groups:control group,model group and chloroquine intervention group.The method to establish calcium oxalate stone model was drinking water with 1％ ethylene and 1％ ammonium chloride freely.The rats of chloroquine intervention group were treat with chloroquine (40mg/kg · d) by intraperitoneal injection.Modeling was finished after 28 days.The amounts of renalcalcium oxalate crystals were detected by polarizing microscope.For all groups,the amounts of autophagosome were detected by transmission electron microscope.Twenty four hour urine compositions for stone risk factors were detected.The expressions of oxidative stress injury related molecular markers (SOD,MCP-1 and 8-OHdG) and the expressions of autophagy markers (LC3 and P62) were detected by immunohistochemistry.The RNA expressions of SLC26A6 in kidney were detected by Real-time PCR.Results Compared to the model group,the amounts of renal calcium oxalate crystals were significantly reduced in chloroquine intervention group (32.37 ± 5.14 vs.4.18 ± 0.25,P ＜ 0.05).Compared to the control group,the level of autophagy was increased in the model group.Compared to the model group,the level of autophagy was inhibited in the chloroquine intervention group.For control group,model group and chloroquine intervention group,the excretion of urinary oxalate were (3.1 ± 1.5) mmol,(22.5 ± 8.1) mmol,(2.8 ± 1.2) mmol,respectively;the excretion of urinary citrate were (63.4 ± 7.4) mmol,(45.9 ± 9.5)mmol,(15.6 ± 8.2) mmol,respectively.Compared to the control group,the amounts of urinary oxalate weresignificantly elevated in model group (P ＜ 0.05),but citrate were significantly reduced in the chloroquineintervention group(P ＜ 0.05).For control group,model group and chloroquine intervention group,theexpressions of SOD were 42.24 ±4.16,19.21 ± 2.25,39.08 3.53,respectively;the expressions of MCP-1 were 4.02 0.51,8.45 ± 0.55,5.52 ± 0.34,respectively;the expressions of 8-OHdG were 7.16 ± 0.54,11.21 ± 1.12,8.67 ±0.34,respectively;the RNA expressions of SLC26A6 were 0.35 ±0.07,1.02 ±0.17,0.70 ± 0.06,respectively.Compared to the control group,the expressions of SOD were significantly reduced in the model group,but the expressions of MCP-1,8-OHdG and SLC26A6 were significantly elevated(P ＜0.05).Compared to the model group,the expressions of SOD were significantly elevated chloroquine intervention group (P ＜ 0.05),but the expressions of MCP-1,8-OHdG and SLC26A6 were significantly elevated(P ＜ 0.05).Conclusions The autophagy inhibitor chloroquine could inhibit the formation of calcium oxalate crystals induced by ethylene in rat kidney via inhibit the renal autophagy level and expressions of the SLC26A6,reducing the renal oxidative stress injury and urinary oxalate excretion.