| Objective:The current incidence of diabetic cystopathy(DCP)is high,the clinical attention is not enough,and its molecular mechanism is not clear.This study investigated the role of cyclooxygenase-2(cox-2)in the development and progression of DCP and its changes after intervention.Method:1.STZ(streptococcus streptococcus)was used to establish the SD rat diabetic bladder model(diabetes group),and urodynamics was used to evaluate the bladder function of the rats,so as to verify the success of DCP modeling;R-PCR,Western blot and immunohistochemistry were used to detect the expression levels of cox-2 and p38-mapk-cox-2 at different levels.2.Eighty rats were divided into five groups:the control group(15rats),diabetes group(12 rats),cox-2 inhibitors(16 rats),P38 lightning inhibitors(16rats),cox-2+P38 lightning inhibitors group(16rats).Western Blot,real-time quantitative PCR(Q-PCR)technique and immunohistochemistry method were used to detect cox-2 and P38 expression level in each group;3.The above results were compared with the results of inhibitor addition and combined with the results of electron microscopy to investigate the role of cox-2 in the development and progression of bladder in diabetes mellitus and the changes after intervention.Results:1.60 diabetic rats were successfully modeled in this experiment,with a success rate of 92%.In the control group,bladder volume(2.23±0.16)ml,bladder pressure(30.67±3.20)cmH2O,and leakage point pressure(59.17±3.13)cmH2O were compared.Bladder volume(4.51±0.32)ml,bladder pressure(24.50±3.55)cmH2O,and leakage point pressure(33.42±6.78)cmH2O in the diabetes group.Compared with the control group,the maximum bladder volume,the maximum bladder pressure,the residual urine volume and the leakage point pressure of rats in the diabetes group were increased,and the differences were statistically significant(P<0.05).2.The levels of related protein molecules in the bladder of rats were detected by Western blot and real-time quantitative PCR:(0.16-0.03)compared with control group,diabetes group(1.30+/-0.11),cox-2 inhibitors group(0.55+/-0.26)and P38 lightning inhibitors(0.96+/-0.12)of cox-2 expression increased significantly,with significant difference statistically significant difference,statistically significant(P<0.01),COX-2+P38 lightning inhibitor group(0.12+0.05)compared with control group,the expression level is lower,the difference is statistically significant(P<0.05),The expression levels of cox-2 inhibitor group,P38 inhibitor group and c0x-2+P38 inhibitor group were significantly lower than that of the diabetes group,with statistically significant differences(P<0.01).The expression levels of cox-2 inhibitor group,cOx-2+P38 inhibitor group were significantly lower than that of the P38 inhibitor group,with statistically significant differences(P<0.01).P38 lightning in diabetic group(1.42+/-0.14),cox-2 inhibitors group(1.42+/-0.30),P38 lightning inhibitors(0.99+0.16),2=+P38 lightning inhibitors COX-group(0.80+0.14)compared with control group(0.53+0.03),significantly increased expression level,with significant difference statistically significant(P<0.01),P38 lightning inhibitors,COX-2+P38 lightning inhibitors group compared with diabetes group,expression level decreased significantly,with significant difference statistically significant(P<0.01),There was no significant change in the expression level between the cox-2 inhibitor group and the diabetes group(P>0.05).3.Immunohistochemical staining intensity comparison;cox-2 expression mainly in the cytoplasm in the urinary bladder,staining intensity respectively:diabetes group(0.74+/-0.14),COX-2 inhibitor group(0.75+/-0.15),P38 lightning inhibitors(0.53+0.16),the control group(0.48+0.38),P38 lightning+cox-2 inhibitors group(0.48+0.22)statistical analysis are as follows:(1)diabetes mellitus group,COX-2 inhibitor,P38 lightning inhibitors group compared with control group were significantly higher(P<0.01);(2)the COX-2 inhibitor group,P38 inhibitor group,P38+cox-2 inhibitor group and diabetes group all significantly reduced(P<0.01).(3)P38 inhibitor group and COX-2 inhibitor group significantly decreased(P<0.05).The cox-2+P38 inhibitor group was significantly lower than the P38 inhibitor group(P<0.01).(4)the difference between the COX-2 inhibitor group and the COX-2+P38 inhibitor group was statistically significant(P<0.01).(5)there was no statistical significance between the P38+COX-2 inhibitor group and the control group(P=0.16>0.05).P38MAPK mainly expressed in the cytoplasm in the urinary bladder,staining intensity respectively:diabetes group(0.74+/-0.14),COX-2 inhibitor group(0.75+/-0.15),P38 lightning inhibitors(0.53+0.16),the control group(0.48+0.38),P38 lightning+ COX-2 inhibitors group(0.48+0.22)statistical analysis are as follows:(1)diabetes mellitus group,COX-2 inhibitor,P38 lightning inhibitors group compared with control group were significantly higher(P<0.01);(2)P38 inhibitor group,P38+COX-2 inhibitor group and diabetes group were significantly reduced(P<0.01).(3)the difference between the P38 inhibitor group and the cox-2 inhibitor group was statistically significant(P=0.03<0.05).(4)the difference between the COX-2 inhibitor group and the COX-2+P38 inhibitor group was statistically significant(P<0.01).(5)compared with the diabetes group,the expression-of P38 in the cOx-2 inhibitor group was not statistically significant(P=0.14>0.05).P38 showed no statistical significance in the P38 inhibitor group and the P38+COX-2 inhibitor group(P=0.53>0.05).4.Observation under light microscope showed that the muscle fibers of the detrusor cross section of the control group were arranged neatly,connected closely,and the nerve structure was complete.A small amount of collagen fibers were filled in the gap.In the DCP group,the muscle fibers were arranged in disorder and the connection structure was disordered.The bladder mucosa was significantly proliferated,and the muscle fibers were fractured to different degrees.The interstitial space was filled with collagen fibers,and the interstitial space was significantly widened.In the COX-2 and p38 inhibitor groups,the detrusor cross section and longitudinal muscle fibers were arranged in a relatively orderly manner,and the connections between them were relatively close.The gap was filled with a small amount of collagen fiber tissue.There was no significant difference between the COX-2 and p38 inhibitor groups.The ultrastructure of mitochondria in the bladder mucosa of diabetic rats was observed to be seriously damaged,which was manifested by the increased mitochondrial volume,the decreased number of "ridges" in the mitochondria,the fracture and even the disappearance,and the decreased light density of mitochondrial matrix.Conclusion;1.In diabetic bladder rats,the maximum bladder volume increased,the maximum bladder pressure decreased,the residual urine increased,and the leakage point pressure decreased.2.Cox-2 and P38 were highly expressed in the bladder of diabetic rats.3.After the use of cox-2 inhibitors,the expression of cox-2 was significantly reduced,but the expression of P38 was not significantly changed.After the use of P38 inhibitor,the expression of P38 decreased significantly,and the expression of cox-2 also decreased significantly.4.This experiment verified that in the p38-cox-2 pathway,P38 is located in the upstream of cox-2 and regulates the expression of cox-2,which has a better inhibitory effect on the two inflammatory factors under the application of dual inhibitors.5.Under the light microscope,detrusor muscle fibers in the diabetes group were observed to be disordered and broken,which indicated that diabetes had an effect on the bladder and damaged the normal physiological structure of detrusor muscle and mucous membrane of the bladder,thus affecting its normal physiological function.6.The damage of mitochondria and other organelles in the bladder mucosa and detrusor cells of the diabetic rats were observed under electron microscope,which affected their functions.The pathological mechanism of the diabetic bladder was described from the perspective of ultrastructure. |
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