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目的探讨糖尿病大鼠膀胱神经生长因子(nerve growth factor,NGF)与糖尿病膀胱病变发生、发展的关系。方法66只SD大鼠随机分为糖尿病组和对照组各33只,糖尿病组腹腔注射链脲佐菌素60mg/kg诱导糖尿病大鼠模型,对照组腹腔注射柠檬酸缓冲液。造模成功后第6、9、12周时2组随机抽取10只大鼠检测体质量和24h尿量,并行尿动力学检测,比较2组最大膀胱容量、残尿量、最大膀胱压力、顺应性和膀胱湿质量;取2组大鼠膀胱组织行HE染色、Masson三色法染色观察膀胱壁组织学变化情况;采用Western blot法检测2组膀胱组织NGF蛋白表达水平,采用反转录PCR法检测2组膀胱组织NGF mRNA表达水平。结果造模第6、9、12周时,糖尿病组大鼠体质量较对照组明显减轻,血糖水平较对照组明显升高,24h尿量较对照组明显增多(P<0.05);造模第6、9、12周时,糖尿病组大鼠最大膀胱容量[(2.25±0.51)、(3.49±0.40)、(4.03±0.68)mL]、顺应性[(1.03±0.13)、(1.39±0.11)、(1.33±0.08)mL/kPa]、膀胱湿质量[(171.15±8.22)、(181.00±9.13)、(183.87±9.55)mg]和残尿量[(0.55±0.47)、(0.66±0.51)、(0.73±0.56)mL]均较对照组[(1.78±0.12)mL、(0.68±0.07)mL/kPa、(135.12±3.54)mg、(0.06±0.01)mL]明显增加(P<0.05);造模第9、12周时糖尿病组最大膀胱压力[(3.09±0.01)、(3.07±0.33)kPa]较对照组[(4.81±0.27)kPa]明显降低(P<0.05);造模第12周时糖尿病组最大膀胱容量、顺应性、膀胱湿质量和残尿量均较第6周时明显增加,最大膀胱压力较第6周时[(4.30±0.24)kPa]明显降低(P<0.05);糖尿病组大鼠第6、9、12周时NGF mRNA表达水平(0.149±0.094、0.109±0.016、0.084±0.078)和蛋白表达水平(0.342±0.056、0.287±0.080、0.106±0.050)均较对照组(0.250±0.123、0.718±0.440)明显降低(P<0.01)。结论 NGF表达水平降低与糖尿病膀胱病变的发生、发展相关。
Objective To investigate the relationship between nerve growth factor (NGF) and the occurrence and development of diabetic bladder disease in diabetic rats. Methods Sixty-six SD rats were randomly divided into diabetic group and control group, with 33 rats in each group. Diabetic rats were induced by intraperitoneal injection of streptozotocin (60mg / kg) and diabetic rats were injected with citrate buffer solution. At the 6th, 9th and 12th week after the model was established, 10 rats in the two groups were randomly selected to detect the body weight and 24h urinary volume, and the urodynamic tests were performed. The maximal bladder capacity, residual urine output and maximal bladder pressure were compared between the two groups Sex and bladder wet weight. Bladder tissue was taken from 2 groups for HE staining and Masson’s trichrome staining for histological changes of bladder wall. Western blot was used to detect the expression of NGF in 2 groups of bladder tissues. Reverse transcription-polymerase chain reaction The expression of NGF mRNA in two groups of bladder tissues was detected. Results At the 6th, 9th and 12th week, the body weight of rats in diabetic group was significantly lower than that in control group, the blood glucose level was significantly higher than that in control group, and the urine output in 24h group was significantly higher than that in control group (P <0.05) At 6, 9 and 12 weeks, the maximum bladder capacity in diabetic rats [(2.25 ± 0.51), (3.49 ± 0.40), (4.03 ± 0.68) mL], compliance [(1.03 ± 0.13), (1.39 ± 0.11) , (1.33 ± 0.08) mL / kPa], bladder wet mass [(171.15 ± 8.22), (181.00 ± 9.13), (183.87 ± 9.55) mg] and residual urine volume (0.55 ± 0.47) and (0.66 ± 0.51) , (0.73 ± 0.56) mL] were significantly higher than those in the control group [(1.78 ± 0.12) mL, (0.68 ± 0.07) mL / kPa, (135.12 ± 3.54) mg and (0.06 ± 0.01) mL] ; Compared with the control group [(4.81 ± 0.27) kPa] (P <0.05), the maximal bladder pressure in the diabetes mellitus group was significantly lower at the 9th and 12th weeks after modeling ([3.09 ± 0.01], (3.07 ± 0.33) kPa] At 12 weeks, the maximum bladder capacity, compliance, bladder wet weight and residual urine volume in diabetic group were significantly increased compared with the 6th week, and the maximum bladder pressure was significantly lower than that of the 6th week [(4.30 ± 0.24) kPa] (P <0.05 ); NGF mRNA expression levels (0.149 ± 0.094, 0.109 ± 0.016, 0.084 ± 0.078) and protein (0.342 ± 0.056,0.287 ± 0.080,0.106 ± 0.050) were significantly lower than those in the control group (0.250 ± 0.123,0.718 ± 0.440) (P <0.01). Conclusion The decreased expression of NGF is associated with the occurrence and development of diabetic bladder disease.