To Explore The Effect Of GPER On Rat Renal Tubular Epithelial Cell Apoptosis After RIRI

Objective: To explore the renal ischemia-reperfusion(ischemia-reperfusion,I/R)injury.G protein-coupled estrogen receptor(GPER)effects on the apoptosis of rat renal tubular epithelial cells and related mechanisms.Methods:(1)32 SPF-grade female Sprague-Dawley(SD)rats were selected.First,all female SD rats were fasted with water for eight hours and then anesthetized,and both sides of the ovaries(ovariectomized,OVX)were surgically removed.After two weeks of routine rearing,they were randomly divided into 4groups with 8 animals in each group.The first group of OVX group;the second group of OVX+I/R group;the third group of OVX+I/R+G1(GPER agonist)group;the fourth group of OVX+I/R+G15(GPER blocker)+G1 group.OVX group: OVX was not treated for regular feeding.OVX+I/R group: Remov the right kidney,and the left renal artery was naked and clamped.After 45 minutes and one day of reperfusion,the OVX rat kidney I/R injury model was successfully prepared for use.OVX+I/R+G1 group: G1intervention(120μg/kg·d)was given by intraperitoneal injection for 3 consecutive days.The rest of the steps were the same as those in the OVX+I/R group to prepare rat kidney I/R injury model.OVX+I/R+G15+G1 group: G15 blocker(300μg/kg·d)was given by intraperitoneal injection,and G1intervention(120μg/kg·d)was given by intraperitoneal injection 30 minutes later,for 3 consecutive days,and the remaining steps were the same as OVX The rat kidney I/R injury model was prepared in the I/R group.Rats in each group were reared routinely for one day after successful modeling,and the left kidney was taken out immediately after blood sampling from the abdominal aorta through a median abdominal incision.(2)Measure the levels of arterial serum creatinine(SCr),arterial serum urea nitrogen(BUN),lactate dehydrogenase(LDH)of rats in each group to evaluate renal function.(3)Kidney tissue hematoxylin-eosin staining,Paller score jointly evaluate the degree of renal tissue damage.(4)Immunohistochemical.technique to detect the.expression position and positive.expression level.of apoptosis-related.protein in kidney tissue.(5)Western-blot.was used to detect and.quantify the expression.level of apoptotic protein in renal tissue.Results:(1)Compared with.the OVX group,the serum SCr,BUN,LDH levels in the.OVX+I/R group were.significantly increased(P<0.01);compared with the.OVX +I/R group,the serum SCr,BUN,and BUN levels in the.G1 intervention group.The level of LDH.decreased significantly(P<0.01);G15intervention can block the intervention effect of G1(P<0.01).(2)Hematoxylin-eosin.staining results:Compared with the.OVX group,the renal tubules.in the OVX+I/R group were.significantly expanded.Some renal tubules were unclear in structure,epithelial cell edema,nuclear anoikis,and lumen obstruction.The appearance of casts,the brush border can be seen to be damaged and shedding,and the Paller score increased,indicating that the renal tubular structure was seriously damaged(P<0.01).After G1 intervention,the renal tubules were slightly dilated,and the cell morphology was reduced compared with I/R group necrosis and shedding.The decrease of Paller score indicated that renal tubule damage was significantly reduced(P<0.01).The G15 blocker group reversed the protective effect of G1 activated GPER(P<0.01).(3)Results of immunohistochemical technique: Compared with the.OVX group,the OVX+I/R group.suppressed the positive expression of protein.Bcl-2 and promoted the positive.expression of protein Caspase-3;compared with the.OVX+I/R group,G1 intervention.In the group,the positive expression of.Bcl-2 protein was obviously promoted,and the positive expression.of Caspase-3 protein was inhibited;at the same time,G15.blocker could reverse.this situation.(4)Results of.Western Blot: Compared with the.OVX group,the OVX+I/R group.reduced the expression.of protein Bcl-2(P<0.01),and significantly.increased the expression of protein Cleaved.Caspase-3(P<0.01).G1 intervention.increased the expression of protein.Bcl-2(P<0.01),and decreased.the expression of.protein Cleaved Caspase-3(P<0.01).The G15 blocker group could partially reverse the intervention effect of G1(P<0.01).Conclusion: GPER can reduce renal I/R injury by reducing the apoptosis of renal tubular epithelial cells,further verifying the protective effect of G1 on renal I/R injury after G1 activates GPER,and its mechanism may be achieved by regulating the apoptosis pathway.