高血压肾交感神经的病理三维重建及组织学和功能学重塑研究

发布时间：2018-05-27 12:10

本文选题：高血压 + 犬肾交感神经　；参考：《重庆医科大学》2017年博士论文

【摘要】：第一部分二维层面高血压犬肾交感神经的组织学和功能学重塑研究目的:在二维层面运用常规病理切片评估高血压犬肾交感神经(Renal sympathetic nerve,RSN)是否存在组织学和功能学重塑。方法:对老年昆明犬进行有创血压测量,根据血压测量结果将其分为高血压犬组(n=7)和非高血压犬组(n=7)。将左侧肾动脉平均分为近、中、远3段行HE染色、酪氨酸羟化酶(tyrosine hydroxylase,TH)和P物质免疫组织化学染色。取左侧主动脉肾神经节行HE染色、TH和生长因子相关蛋白43(Growth associated protein 43,GAP43)免疫组织化学染色。取新鲜肾脏组织行高效液相色谱(High performance liquid chromatography,HPLC)分析,测量肾组织去甲肾上腺素浓度(norepinephrine,NE)。结果:高血压组的平均血压(175.7±8.5/116.0±9.1 mm Hg)显著高于非高血压组(124.1±9.8/85.4±5.1 mm Hg,P0.01)。高血压组与非高血压组相比,肾动脉近段的RSN数目(P=0.042)显著增多,但在肾动脉中段(P=0.417)和远段(P=0.641)二者无显著差异。肾动脉各节段RSN的大小和距肾动脉内膜距离在两组间均无统计学差异。高血压组TH(P=0.025)和P物质(P0.01)的平均密度以及肾组织NE浓度(P=0.027)均显著高于非高血压组。主动脉肾神经节神经元的平均面积在高血压组要显著高于非高血压组(P=0.023),但两组每个高倍视野下的平均神经元数目相当(P=0.21)。高血压时神经元GAP43表达显著增强(P=0.012),TH的表达也略高于非高血压组,但并无统计学差异(P=0.156)。结论:高血压时主动脉肾神经节可能存在组织学和功能学重塑,且RSN传入和传神经纤维的功能也有不同程度增强。但在高血压时RSN是否存在组织学重塑还需在三维层面进一步研究。第二部分高血压与非高血压犬肾交感神经的病理三维重建目的:利用病理三维重建技术(Pathological three-dimensional reconstruction,PTDR)三维重建犬肾交感神经(Renal sympathetic nerve,RSN),在三维层面分析RSN的解剖学特征并进一步明确高血压犬的RSN是否存在组织学重塑。方法:取犬右侧肾动脉(存在副肾动脉时取左侧)并以女性发丝作为定位标识,对肾动脉及其周围组织行间断连续冰冻切片和HE染色。利用高清扫描仪获取切片病理图像,并利用Photoshop软件根据定位标识进行图像匹配。将所有匹配图像导入3D-doctor软件,根据图像大小设置三维重建参数,对不同组织进行轮廓分割,最终采用Complex surface模式重建犬RSN的三维图像。利用3D-doctor软件将图像平均分为近段、中段和远段三个部分,自动计算肾动脉、RSN的体积等参数,手动完成RSN的计数和距离测量。结果:利用3D-doctor软件成功重建出犬RSN的三维图像。犬的RSN多呈单根形态从肾动脉上极发出,缠绕肾动脉并与其伴行至肾动脉远端入肾。犬肾动脉近段RSN的体积要大于中段(P0.01)和远段(P0.01)。RSN在上极(P0.01)的数目多于下极、腹侧和背侧,但近、中、远3段的RSN数目无显著差异(P=0.816)。RSN从肾动脉近段行进至中段和远段的过程中距肾血管内膜的距离逐渐减小(P0.01)。总体上,高血压犬RSN的数目要多于非高血压犬(P=0.02)。但在RSN距肾动脉内膜距离(P=0.054),RSN平均体积(P=0.12)和RSN/肾动脉体积比(P=0.44)方面,二者并无显著差异。结论:本部分研究首次利用PTDR技术重建出犬RSN的三维图像,并在三维层面对犬RSN的解剖学特征进行了较为详尽的描述。此外,我们发现高血压犬RSN的数目多于非高血压犬,提示高血压可能导致RSN的组织学重塑。第三部分人体肾交感神经的病理三维重建及组织学和功能学重塑目的:三维重建人体肾交感神经(Renal sympathetic nerve,RSN),评估其三维解剖分布特征;比较高血压和非高血压人群RSN和肾神经节(Renal ganglion,RG)的三维图像和免疫组化特征,明确高血压时人体RSN和RG是否存在组织学和功能学重塑。方法:收集新鲜人体肾动脉标本进行间断连续冰冻切片,利用病理三维重建技术重建人体肾交感神经(Renal sympathetic nerve,RSN)。测量RSN体积、RG体积、RSN和肾动脉体积比、RG和肾动脉体积比、RSN距肾动脉内膜最大距离、RSN距肾动脉内膜最小距离。对人体RSN冰冻切片行酪氨酸羟化酶(tyrosine hydroxylase,TH)和P物质免疫组化染色。对人体RG冰冻切片行TH和生长因子相关蛋白43(Growth associated protein 43,GAP43)免疫组化染色。收集患者病史资料,比较高血压与非高血压人群上述参数是否存在差异。结果:本部分研究纳入7例尸检病例(高血压4例,非高血压3例)共11条肾动脉(左侧5条,右侧6条)。三维图像显示人体RSN在肾动脉周围呈网状分布。肾动脉近段RSN平均体积和RSN/肾动脉体积比要大于中段和远段。肾动脉近段和中段的RG体积以及RG/肾动脉体积比显著大于远段。人体RSN从肾动脉近段行至远段的过程中,距肾动脉内膜的距离逐渐减小。高血压患者RSN体积与非高血压患者相当(P=0.072),但二者RSN/肾动脉体积比存在显著差异(P=0.01)。RG体积(P=0.029)和RG/肾动脉体积比(P=0.045)在高血压患者显著增大。RSN TH的平均密度在高血压组要显著高于非高血压组(P0.01),但P物质的平均密度在两组并无显著差异(P=0.80)。此外,RG平均神经元面积(P0.01)和GAP43的表达(P0.01)在高血压人群中也显著增加。结论:肾动脉近段RSN和RG的体积较大,但离肾动脉距离较远;虽然远段RSN体积较小,但离肾动脉距离较近。因此选择在肾动脉远段进行消融可能有助于提高肾去交感术的有效性。此外,本研究发现高血压患者RSN传出神经纤维的功能增强,RG体积增大,RG功能也显著增强,提示高血压患者RG和RSN存在不同程度的组织学和功能学重塑。
[Abstract]:Part 1: Study on histological and functional remodeling of renal sympathetic nerve in two dimensional hypertensive dogs. Objective: To evaluate the histological and functional remodeling of Renal sympathetic nerve (RSN) in hypertensive dogs by routine pathological sections at two-dimensional level. Methods: invasive blood pressure measurements were carried out in old Kunming dogs, according to blood pressure. The results were divided into the hypertension dog group (n=7) and the non hypertensive dog group (n=7). The left renal artery was divided into the proximal, middle and far 3 segments with HE staining, tyrosine hydroxylase (tyrosine hydroxylase, TH) and P substance immunohistochemical staining. The left aorta renal node was stained with HE, TH and growth factor related protein 43 (Growth associated P). Rotein 43, GAP43) immunohistochemical staining. Analysis of High performance liquid chromatography (HPLC) in fresh renal tissue and measurement of renal tissue norepinephrine concentration (norepinephrine, NE). Results: the mean blood pressure (175.7 + 8.5/116.0 + 9.1 mm Hg) in the hypertensive group was significantly higher than that in the non hypertensive group (124.1 +. 4 + 5.1 mm Hg, P0.01). Compared with non hypertension group, the number of RSN (P=0.042) in the proximal renal artery increased significantly, but there was no significant difference between the two renal artery (P=0.417) and the distal segment (P=0.641). The size of RSN in each segment of the renal artery and the distance from the renal artery intima were not statistically different. The TH (P=0.025) and P substance in the hypertension group The mean density of P0.01 and the concentration of NE in the renal tissue (P=0.027) were significantly higher than those in the non hypertensive group. The mean area of the aorta renal ganglion neurons in the hypertensive group was significantly higher than that in the non hypertensive group (P=0.023), but the average number of neurons in the two groups was equal to that in the high field of vision (P=0.21). The expression of GAP43 in the neuron was significantly increased in hypertension. The expression of strong (P=0.012), TH is also slightly higher than that in non hypertension group, but there is no statistical difference (P=0.156). Conclusion: the aorta and renal ganglion may have histological and functional remodeling in hypertension, and the function of RSN afferent and transmission of nerve fiber is also enhanced in varying degrees. But in high blood pressure, the existence of tissue remodeling in RSN needs to be in the three-dimensional level. Further study. The pathological three dimensional reconstruction of renal sympathetic nerve in second parts of hypertension and non hypertensive dogs: three-dimensional reconstruction of the renal sympathetic nerve (Renal sympathetic nerve, RSN) by Pathological three-dimensional reconstruction (PTDR). The anatomical features of RSN were analyzed and further demonstrated in the three-dimensional level. Methods: whether there is a tissue remodeling in the RSN of a dog with high blood pressure. Method: Taking the right renal artery in the dog (left renal artery in the left) and using a female hairline as a positioning sign, continuous frozen section and HE staining on the intersections of the renal artery and its surrounding tissue. The pathological image of the slice was obtained by the high-definition scanner, and the Photoshop software was used to locate the pathological images. The image is matched by the logo. All the matched images are imported into 3D-doctor software, the 3D reconstruction parameters are set according to the size of the image, the contour of different organizations is segmented, and the three-dimensional image of the dog RSN is reconstructed by the Complex surface model. The image is divided into three parts, the near segment, the middle and the far segments, with the 3D-doctor software to automatically calculate the kidney. The volume and other parameters of the artery, RSN volume and other parameters were done manually. Results: the three-dimensional image of the dog's RSN was successfully reconstructed by the 3D-doctor software. The dog's RSN was mostly emitted from the renal artery, and the renal artery was twined and accompanied with the renal artery to the distal part of the kidney. The volume of the near segment of the dog's renal artery was larger than the middle segment (P0.01) and far away from the middle segment of the renal artery (P0.01) and far away from the dog's renal artery. The number of segment (P0.01).RSN in the upper pole (P0.01) was more than the lower pole, the ventral side and the dorsal side, but the number of RSN in the near, middle and far 3 segments had no significant difference (P=0.816).RSN from the proximal renal artery to the middle and distal segments gradually decreased from the renal vascular intima (P0.01). In general, the number of hypertensive dogs RSN was more than that of non hypertensive dogs (P=0.02). There was no significant difference between the RSN distance (P=0.054), the mean volume of RSN (P=0.12) and the RSN/ renal artery volume ratio (P=0.44). Conclusion: this part of this study was the first to reconstruct the three-dimensional images of the dog RSN by PTDR technique and to describe the anatomical features of the dog RSN in a more detailed description in the three-dimensional layer. In addition, we found that the two The number of RSN in hypertensive dogs is more than that of non hypertensive dogs. It is suggested that hypertension may lead to the histologic remodeling of RSN. The third part of the pathological reconstruction of the human renal sympathetic nerve and the remodeling of histology and function: three-dimensional reconstruction of the Renal sympathetic nerve (RSN) in the human body, to evaluate the three-dimensional anatomical distribution of the human kidney, and to compare the hypertension. The three-dimensional images and immunohistochemical features of RSN and Renal ganglion (RG) in the non hypertensive population, and to determine whether there is a histological and functional remodeling of RSN and RG in human body during hypertension. Methods: collecting fresh human renal artery specimens for continuous frozen section and reconstructing the human renal sympathetic nerve (Ren) with pathological three-dimensional reconstruction technique. Al sympathetic nerve, RSN). Measurement of RSN volume, RG volume, RSN and renal artery volume ratio, RG and renal artery volume ratio, RSN distance from the renal artery intima, RSN distance from the renal artery intima. Growth factor related protein 43 (Growth associated protein 43, GAP43) immunohistochemical staining. Collect patients' medical history data to compare the differences between the above parameters of hypertension and non hypertensive people. Results: this part of the study included 7 cases of autopsy (4 cases of hypertension, 3 cases of non high blood pressure) and 11 renal arteries (left 5, right 6). The average volume of RSN in the proximal part of the renal artery and the volume ratio of RSN/ to the renal artery were greater than that of the middle and distal segments. The RG volume and the volume ratio of RG/ renal artery in the proximal and middle segments of the renal artery were significantly greater than that in the far segment. The distance from the human RSN from the proximal renal artery to the distal part of the renal artery was gradually greater than that of the distal segment of the renal artery. The RSN volume of hypertensive patients was equivalent to that of non hypertensive patients (P=0.072), but there was a significant difference in the volume ratio of RSN/ renal arteries in the two (P=0.01).RG volume (P=0.029) and RG/ renal artery volume ratio (P=0.045) in hypertensive patients, the average density of.RSN TH was significantly higher in the high blood pressure group than in the non hypertensive group (P0.01), but the P substance was flat. There was no significant difference in mean density between the two groups (P=0.80). In addition, the mean neuron area of RG (P0.01) and the expression of GAP43 (P0.01) also increased significantly in the hypertensive population. Conclusion: the volume of the proximal RSN and RG in the renal arteries is larger, but the distance from the renal artery is far away; although the distal RSN volume is smaller, the distance from the renal artery is close. Therefore, the choice of the renal artery is in the renal artery. Ablation in the far segment may help to improve the effectiveness of renal hyperactivity. In addition, this study found that the function of RSN efferent nerve fibers in hypertensive patients was enhanced, the volume of RG increased, and the function of RG was significantly enhanced, suggesting that RG and RSN in hypertensive patients have different degrees of histology and energy remodelling.
【学位授予单位】：重庆医科大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R544.1

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