本文選題：高血壓 + 犬腎交感神經(jīng)��； 參考：《重慶醫(yī)科大學(xué)》2017年博士論文

【摘要】：第一部分二維層面高血壓犬腎交感神經(jīng)的組織學(xué)和功能學(xué)重塑研究目的:在二維層面運(yùn)用常規(guī)病理切片評(píng)估高血壓犬腎交感神經(jīng)(Renal sympathetic nerve,RSN)是否存在組織學(xué)和功能學(xué)重塑。方法:對(duì)老年昆明犬進(jìn)行有創(chuàng)血壓測(cè)量,根據(jù)血壓測(cè)量結(jié)果將其分為高血壓犬組(n=7)和非高血壓犬組(n=7)。將左側(cè)腎動(dòng)脈平均分為近、中、遠(yuǎn)3段行HE染色、酪氨酸羥化酶(tyrosine hydroxylase,TH)和P物質(zhì)免疫組織化學(xué)染色。取左側(cè)主動(dòng)脈腎神經(jīng)節(jié)行HE染色、TH和生長(zhǎng)因子相關(guān)蛋白43(Growth associated protein 43,GAP43)免疫組織化學(xué)染色。取新鮮腎臟組織行高效液相色譜(High performance liquid chromatography,HPLC)分析,測(cè)量腎組織去甲腎上腺素濃度(norepinephrine,NE)。結(jié)果:高血壓組的平均血壓(175.7±8.5/116.0±9.1 mm Hg)顯著高于非高血壓組(124.1±9.8/85.4±5.1 mm Hg,P0.01)。高血壓組與非高血壓組相比,腎動(dòng)脈近段的RSN數(shù)目(P=0.042)顯著增多,但在腎動(dòng)脈中段(P=0.417)和遠(yuǎn)段(P=0.641)二者無(wú)顯著差異。腎動(dòng)脈各節(jié)段RSN的大小和距腎動(dòng)脈內(nèi)膜距離在兩組間均無(wú)統(tǒng)計(jì)學(xué)差異。高血壓組TH(P=0.025)和P物質(zhì)(P0.01)的平均密度以及腎組織NE濃度(P=0.027)均顯著高于非高血壓組。主動(dòng)脈腎神經(jīng)節(jié)神經(jīng)元的平均面積在高血壓組要顯著高于非高血壓組(P=0.023),但兩組每個(gè)高倍視野下的平均神經(jīng)元數(shù)目相當(dāng)(P=0.21)。高血壓時(shí)神經(jīng)元GAP43表達(dá)顯著增強(qiáng)(P=0.012),TH的表達(dá)也略高于非高血壓組,但并無(wú)統(tǒng)計(jì)學(xué)差異(P=0.156)。結(jié)論:高血壓時(shí)主動(dòng)脈腎神經(jīng)節(jié)可能存在組織學(xué)和功能學(xué)重塑,且RSN傳入和傳神經(jīng)纖維的功能也有不同程度增強(qiáng)。但在高血壓時(shí)RSN是否存在組織學(xué)重塑還需在三維層面進(jìn)一步研究。第二部分高血壓與非高血壓犬腎交感神經(jīng)的病理三維重建目的:利用病理三維重建技術(shù)(Pathological three-dimensional reconstruction,PTDR)三維重建犬腎交感神經(jīng)(Renal sympathetic nerve,RSN),在三維層面分析RSN的解剖學(xué)特征并進(jìn)一步明確高血壓犬的RSN是否存在組織學(xué)重塑。方法:取犬右側(cè)腎動(dòng)脈(存在副腎動(dòng)脈時(shí)取左側(cè))并以女性發(fā)絲作為定位標(biāo)識(shí),對(duì)腎動(dòng)脈及其周圍組織行間斷連續(xù)冰凍切片和HE染色。利用高清掃描儀獲取切片病理圖像,并利用Photoshop軟件根據(jù)定位標(biāo)識(shí)進(jìn)行圖像匹配。將所有匹配圖像導(dǎo)入3D-doctor軟件,根據(jù)圖像大小設(shè)置三維重建參數(shù),對(duì)不同組織進(jìn)行輪廓分割,最終采用Complex surface模式重建犬RSN的三維圖像。利用3D-doctor軟件將圖像平均分為近段、中段和遠(yuǎn)段三個(gè)部分,自動(dòng)計(jì)算腎動(dòng)脈、RSN的體積等參數(shù),手動(dòng)完成RSN的計(jì)數(shù)和距離測(cè)量。結(jié)果:利用3D-doctor軟件成功重建出犬RSN的三維圖像。犬的RSN多呈單根形態(tài)從腎動(dòng)脈上極發(fā)出,纏繞腎動(dòng)脈并與其伴行至腎動(dòng)脈遠(yuǎn)端入腎。犬腎動(dòng)脈近段RSN的體積要大于中段(P0.01)和遠(yuǎn)段(P0.01)。RSN在上極(P0.01)的數(shù)目多于下極、腹側(cè)和背側(cè),但近、中、遠(yuǎn)3段的RSN數(shù)目無(wú)顯著差異(P=0.816)。RSN從腎動(dòng)脈近段行進(jìn)至中段和遠(yuǎn)段的過(guò)程中距腎血管內(nèi)膜的距離逐漸減小(P0.01)�？傮w上,高血壓犬RSN的數(shù)目要多于非高血壓犬(P=0.02)。但在RSN距腎動(dòng)脈內(nèi)膜距離(P=0.054),RSN平均體積(P=0.12)和RSN/腎動(dòng)脈體積比(P=0.44)方面,二者并無(wú)顯著差異。結(jié)論:本部分研究首次利用PTDR技術(shù)重建出犬RSN的三維圖像,并在三維層面對(duì)犬RSN的解剖學(xué)特征進(jìn)行了較為詳盡的描述。此外,我們發(fā)現(xiàn)高血壓犬RSN的數(shù)目多于非高血壓犬,提示高血壓可能導(dǎo)致RSN的組織學(xué)重塑。第三部分人體腎交感神經(jīng)的病理三維重建及組織學(xué)和功能學(xué)重塑目的:三維重建人體腎交感神經(jīng)(Renal sympathetic nerve,RSN),評(píng)估其三維解剖分布特征;比較高血壓和非高血壓人群RSN和腎神經(jīng)節(jié)(Renal ganglion,RG)的三維圖像和免疫組化特征,明確高血壓時(shí)人體RSN和RG是否存在組織學(xué)和功能學(xué)重塑。方法:收集新鮮人體腎動(dòng)脈標(biāo)本進(jìn)行間斷連續(xù)冰凍切片,利用病理三維重建技術(shù)重建人體腎交感神經(jīng)(Renal sympathetic nerve,RSN)。測(cè)量RSN體積、RG體積、RSN和腎動(dòng)脈體積比、RG和腎動(dòng)脈體積比、RSN距腎動(dòng)脈內(nèi)膜最大距離、RSN距腎動(dòng)脈內(nèi)膜最小距離。對(duì)人體RSN冰凍切片行酪氨酸羥化酶(tyrosine hydroxylase,TH)和P物質(zhì)免疫組化染色。對(duì)人體RG冰凍切片行TH和生長(zhǎng)因子相關(guān)蛋白43(Growth associated protein 43,GAP43)免疫組化染色。收集患者病史資料,比較高血壓與非高血壓人群上述參數(shù)是否存在差異。結(jié)果:本部分研究納入7例尸檢病例(高血壓4例,非高血壓3例)共11條腎動(dòng)脈(左側(cè)5條,右側(cè)6條)。三維圖像顯示人體RSN在腎動(dòng)脈周圍呈網(wǎng)狀分布。腎動(dòng)脈近段RSN平均體積和RSN/腎動(dòng)脈體積比要大于中段和遠(yuǎn)段。腎動(dòng)脈近段和中段的RG體積以及RG/腎動(dòng)脈體積比顯著大于遠(yuǎn)段。人體RSN從腎動(dòng)脈近段行至遠(yuǎn)段的過(guò)程中,距腎動(dòng)脈內(nèi)膜的距離逐漸減小。高血壓患者RSN體積與非高血壓患者相當(dāng)(P=0.072),但二者RSN/腎動(dòng)脈體積比存在顯著差異(P=0.01)。RG體積(P=0.029)和RG/腎動(dòng)脈體積比(P=0.045)在高血壓患者顯著增大。RSN TH的平均密度在高血壓組要顯著高于非高血壓組(P0.01),但P物質(zhì)的平均密度在兩組并無(wú)顯著差異(P=0.80)。此外,RG平均神經(jīng)元面積(P0.01)和GAP43的表達(dá)(P0.01)在高血壓人群中也顯著增加。結(jié)論:腎動(dòng)脈近段RSN和RG的體積較大,但離腎動(dòng)脈距離較遠(yuǎn);雖然遠(yuǎn)段RSN體積較小,但離腎動(dòng)脈距離較近。因此選擇在腎動(dòng)脈遠(yuǎn)段進(jìn)行消融可能有助于提高腎去交感術(shù)的有效性。此外,本研究發(fā)現(xiàn)高血壓患者RSN傳出神經(jīng)纖維的功能增強(qiáng),RG體積增大,RG功能也顯著增強(qiáng),提示高血壓患者RG和RSN存在不同程度的組織學(xué)和功能學(xué)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.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R544.1

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