CNP对Ach引起的糖尿病大鼠胃平滑肌收缩增强的影响及IGF-1的干预效应
发布时间：2018-11-02 19:04
【摘要】：目的：本实验通过观察C型钠尿肽、乙酰胆碱、硝普钠等药物对STZ诱导的糖尿病模型大鼠胃自发性收缩活动的作用以及IGF-1对糖尿病胃动力障碍的治疗效果,进一步探讨糖尿病胃动力障碍的发病机理,以期为寻找防治糖尿病胃动力障碍的有效方案提供理论依据。方法：Wistar雄性大鼠30只(250g±20g),随机分为正常对照组、模型组、IGF-1治疗组各10只。STZ腹腔注射制备糖尿病大鼠模型,糖尿病造模成功后第10周对IGF-1治疗组行腹腔注射IGF-1(1500ng/kg)治疗2周,均于造模12周后进行如下实验。测量体质量、血糖、尿量；通过多道生理信号记录系统记录各组大鼠胃窦平滑肌自发性收缩活动；对各组大鼠胃窦平滑肌组织进行HE染色以观察其形态结构；采用ELISA测定血清CNP的含量；利用RT-PCR观察胃组织中CNP mRNA的表达；免疫组织化学方法观察各组大鼠胃平滑肌组织中NPR-A和NPR-B的分布；利用电镜观察胃窦平滑肌细胞内超微结构；利用Western bolt观察胃平滑肌组织中GRP78的表达。结果:1.模型组及IGF-1治疗组大鼠血糖浓度明显高于正常对照组大鼠(n=6,P0.01),体重明显低于正常对照组大鼠(n=6,P0.05),尿量明显多于正常对照组大鼠(n=6,P0.01),差异均有统计学意义;2.HE染色示模型组及IGF-1治疗组大鼠胃窦平滑肌变薄；IGF-1治疗组与模型组间无显著性差异。3.模型组及IGF-1治疗组大鼠胃收缩张力、频率较正常组明显减低,对乙酰胆碱敏感性降低,对CNP、SNP敏感性增加,差异均有统计学意义(n=6,p0.05)。4.免疫组织化学结果显示模型组及IGF-1治疗组大鼠与正常对照组相比胃窦平滑肌组织NPR-B表达明显增多,差异具有统计学意义(n=9,p0.05),各组大鼠胃平滑肌中NPR-A表达无显著性差异。5.IGF-1治疗组大鼠胃肌条收缩、胃平滑肌形态学改变、胃平滑肌组织NPR-B、 NPR-A较模型组无显著性差异(n=6,P0.05)。6. ELISA结果显示模型组及IGF-1治疗组大鼠血清中的CNP较正常对照组无显著性差异(n=6,P0.05)。7. RT-PCR结果显示模型组及工GF-1治疗组大鼠胃窦平滑肌组织中的CNPmRNA表达较正常组明显增多,差异具有统计学意义(n=6,P0.05)。8.电镜下可见,与对照组相比,模型组及IGF-1治疗组平滑肌细胞内质网明显肿胀、空泡化,线粒体极消失,但IGF-1治疗组较模型组明显改善(n=6,P0.05)。9. Western结果显示IGF-1治疗组GRP78表达明显低于模型组,差异具有统计学意义(n=6,P0.05)。结论：STZ诱导的糖尿病大鼠发病12周后,胃窦平滑肌自发性收缩活动明显异常,收缩频率减慢、振幅明显下降、收缩节律紊乱；内源性CNP参与糖尿病胃动力障碍大鼠胃收缩的调节；胃窦平滑肌NP-NPR-B-cGMP和NO-sGC-cGMP信号转导途径的活性明显提高；IGF-1治疗对糖尿病胃动力障碍大鼠胃收缩活动无明显改善作用,但是可以阻止或改善内质网应激作用。
[Abstract]:Objective: to observe the effect of type C natriuretic peptide, acetylcholine and sodium nitroprusside on gastric spontaneous contraction induced by STZ in diabetic rats and the therapeutic effect of IGF-1 on diabetic gastric motility. To explore the pathogenesis of diabetic gastric motility disorder in order to provide theoretical basis for finding an effective scheme for prevention and treatment of diabetic gastric motility disorder. Methods: thirty Wistar male rats (250g 卤20g) were randomly divided into normal control group, model group and IGF-1 treatment group. Diabetic rats were induced by intraperitoneal injection of STZ. The IGF-1 group was treated with intraperitoneal injection of IGF-1 (1500ng/kg) for 2 weeks at the 10th week after successful diabetic modeling. The following experiments were carried out 12 weeks later. Body mass, blood glucose and urine volume were measured. Spontaneous contraction of gastric antral smooth muscle was recorded by multi-channel physiological signal recording system. The antral smooth muscle tissue of each group was stained with HE to observe its morphological structure. The content of serum CNP was measured by ELISA, the expression of CNP mRNA in gastric tissue was observed by RT-PCR, the distribution of NPR-A and NPR-B in gastric smooth muscle tissue was observed by immunohistochemical method. The ultrastructure of gastric antral smooth muscle cells and the expression of GRP78 in gastric smooth muscle tissue were observed by electron microscope and Western bolt respectively. The result is 1: 1. The blood glucose concentration in the model group and the IGF-1 group was significantly higher than that in the normal control group (n = 6, P 0.01), and the body weight was significantly lower than that in the normal control group (P 0.05), and the urine volume was significantly higher than that in the normal control group (n = 6, P 0.01). The differences were statistically significant. 2.HE staining showed that gastric antrum smooth muscle thinned in model group and IGF-1 group. There was no significant difference between IGF-1 treatment group and model group. The gastric contractile tension and frequency in model group and IGF-1 group were significantly lower than those in normal group, and the sensitivity to acetylcholine was decreased, and the sensitivity to CNP,SNP was increased, with significant difference (nong6, p0.05). 4. The results of immunohistochemistry showed that the expression of NPR-B in the antral smooth muscle tissue of the model group and the IGF-1 group was significantly higher than that of the normal control group, and the difference was statistically significant. There was no significant difference in the expression of NPR-A in the gastric smooth muscle of rats in each group. The gastric muscle strips contracted and the gastric smooth muscle morphologically changed in the 5.IGF-1 treatment group. There was no significant difference in NPR-B, NPR-A in the gastric smooth muscle tissue compared with the model group (n = 6, P < 0.05). P0.05). 6. ELISA results showed that there was no significant difference in serum CNP between the model group and the IGF-1 treatment group compared with the normal control group (7. 05). The results of RT-PCR showed that the expression of CNPmRNA in gastric antral smooth muscle tissue in model group and GF-1 group was significantly higher than that in normal group (nti6P 0.05). Compared with the control group, the endoplasmic reticulum of smooth muscle cells in the model group and IGF-1 treatment group was obviously swollen, vacuolated and mitochondria disappeared, but the IGF-1 group was significantly improved compared with the model group (P 0.05). Western results showed that the expression of GRP78 in IGF-1 group was significantly lower than that in model group (P 0.05). Conclusion: the spontaneous contractile activity of gastric antral smooth muscle in diabetic rats induced by STZ was obviously abnormal, the contractile frequency slowed down, the amplitude decreased, and the contraction rhythm was disturbed 12 weeks after onset. Endogenous CNP was involved in the regulation of gastric contraction in diabetic rats with gastric motility disorder, and the activity of NP-NPR-B-cGMP and NO-sGC-cGMP signal transduction pathway in gastric antral smooth muscle was significantly increased. IGF-1 treatment did not significantly improve gastric contractile activity in diabetic rats with gastric motility disorder, but could prevent or improve endoplasmic reticulum stress.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.2
本文编号：2306684
[Abstract]:Objective: to observe the effect of type C natriuretic peptide, acetylcholine and sodium nitroprusside on gastric spontaneous contraction induced by STZ in diabetic rats and the therapeutic effect of IGF-1 on diabetic gastric motility. To explore the pathogenesis of diabetic gastric motility disorder in order to provide theoretical basis for finding an effective scheme for prevention and treatment of diabetic gastric motility disorder. Methods: thirty Wistar male rats (250g 卤20g) were randomly divided into normal control group, model group and IGF-1 treatment group. Diabetic rats were induced by intraperitoneal injection of STZ. The IGF-1 group was treated with intraperitoneal injection of IGF-1 (1500ng/kg) for 2 weeks at the 10th week after successful diabetic modeling. The following experiments were carried out 12 weeks later. Body mass, blood glucose and urine volume were measured. Spontaneous contraction of gastric antral smooth muscle was recorded by multi-channel physiological signal recording system. The antral smooth muscle tissue of each group was stained with HE to observe its morphological structure. The content of serum CNP was measured by ELISA, the expression of CNP mRNA in gastric tissue was observed by RT-PCR, the distribution of NPR-A and NPR-B in gastric smooth muscle tissue was observed by immunohistochemical method. The ultrastructure of gastric antral smooth muscle cells and the expression of GRP78 in gastric smooth muscle tissue were observed by electron microscope and Western bolt respectively. The result is 1: 1. The blood glucose concentration in the model group and the IGF-1 group was significantly higher than that in the normal control group (n = 6, P 0.01), and the body weight was significantly lower than that in the normal control group (P 0.05), and the urine volume was significantly higher than that in the normal control group (n = 6, P 0.01). The differences were statistically significant. 2.HE staining showed that gastric antrum smooth muscle thinned in model group and IGF-1 group. There was no significant difference between IGF-1 treatment group and model group. The gastric contractile tension and frequency in model group and IGF-1 group were significantly lower than those in normal group, and the sensitivity to acetylcholine was decreased, and the sensitivity to CNP,SNP was increased, with significant difference (nong6, p0.05). 4. The results of immunohistochemistry showed that the expression of NPR-B in the antral smooth muscle tissue of the model group and the IGF-1 group was significantly higher than that of the normal control group, and the difference was statistically significant. There was no significant difference in the expression of NPR-A in the gastric smooth muscle of rats in each group. The gastric muscle strips contracted and the gastric smooth muscle morphologically changed in the 5.IGF-1 treatment group. There was no significant difference in NPR-B, NPR-A in the gastric smooth muscle tissue compared with the model group (n = 6, P < 0.05). P0.05). 6. ELISA results showed that there was no significant difference in serum CNP between the model group and the IGF-1 treatment group compared with the normal control group (7. 05). The results of RT-PCR showed that the expression of CNPmRNA in gastric antral smooth muscle tissue in model group and GF-1 group was significantly higher than that in normal group (nti6P 0.05). Compared with the control group, the endoplasmic reticulum of smooth muscle cells in the model group and IGF-1 treatment group was obviously swollen, vacuolated and mitochondria disappeared, but the IGF-1 group was significantly improved compared with the model group (P 0.05). Western results showed that the expression of GRP78 in IGF-1 group was significantly lower than that in model group (P 0.05). Conclusion: the spontaneous contractile activity of gastric antral smooth muscle in diabetic rats induced by STZ was obviously abnormal, the contractile frequency slowed down, the amplitude decreased, and the contraction rhythm was disturbed 12 weeks after onset. Endogenous CNP was involved in the regulation of gastric contraction in diabetic rats with gastric motility disorder, and the activity of NP-NPR-B-cGMP and NO-sGC-cGMP signal transduction pathway in gastric antral smooth muscle was significantly increased. IGF-1 treatment did not significantly improve gastric contractile activity in diabetic rats with gastric motility disorder, but could prevent or improve endoplasmic reticulum stress.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.2
【参考文献】
相关期刊论文 前5条
1 程晓雯;郑清华;李小玲;林中;;血管活性肠肽与某些胃肠动力紊乱性疾病的内在关联研究进展[J];中国全科医学;2012年03期
2 邢德刚;张燕;梁艳玲;杨红;;钙离子在CNP抑制Ach增强胃窦环行平滑肌收缩效应中的作用[J];中国老年学杂志;2010年14期
3 ;Comparative study in the effect of C-type natriuretic peptide on gastric motility in various animals[J];World Journal of Gastroenterology;2003年03期
4 Xia Chen;Xiang-Sheng Fu;Chang-Ping Li;Hong-Xian Zhao;;ER stress and ER stress-induced apoptosis are activated in gastric SMCs in diabetic rats[J];World Journal of Gastroenterology;2014年25期
5 景琪;吴国泰;杜丽东;刘五州;李应东;任远;;当归黄芪醇提物对糖尿病胃轻瘫模型大鼠的治疗作用[J];中药材;2014年08期
，本文编号：2306684
本文链接：https://www.wllwen.com/yixuelunwen/nfm/2306684.html
最近更新
教材专著
