羟乙基淀粉对兔腹部大手术后早期微血管白蛋白渗漏影响机制的研究

发布时间：2018-04-21 23:08

本文选题：腹部手术 + 血清白蛋白　；参考：《昆明医科大学》2015年硕士论文

【摘要】：目的：通过观察兔腹部大手术后早期肠系膜微血管白蛋白渗漏现象,羟乙基淀粉干预,以及肠系膜微血管内皮细胞间隙的变化,测定肠系膜微血管白蛋白和羟乙基淀粉渗漏率,以及血清白蛋白浓度及血清炎性介质浓度的变化,探讨羟乙基淀粉对兔腹部大手术后早期微血管白蛋白渗漏的影响机制。方法：将54只日本大耳兔随机分为术前对照组6只,术后生理盐水组24只(6h、24h、48h、72h各6只)和术后羟乙基淀粉组24只(6h、24h、48h、72h各6只)。两手术组均行胃大部切除加胰体尾、脾切除术,术后所有兔均采用每公斤体重等热卡、等氮量营养支持,热量60kca/Kg,由葡萄糖及脂肪提供,分别提供热量的70%、30%。氮源由复方氨基酸提供,氮热比为1：130。此外,生理盐水组术后每天给予25ml/Kg的生理盐水至观测时间点；羟乙基淀粉组术后每天给予6%HES (130,0.4) 25ml/kg至观测时间点；对照组(不手术)作为术前检测点,生理盐水组及羟乙基淀粉组以术后(6h、24h、48h、72h)作为检测点,予各检测点检测血清白蛋白浓度、血清白细胞介素-6(Interleukin-6, IL-6)、肿瘤坏死因子-α (Tumour necrosis factor-α, TNF-α)、白细胞介素-2(Interleukin-2, IL-2)、细胞间黏附分子-1(intercellular adhesion molecule-1, ICAM-1)、血管内皮生长因子(vascular endothelial growth factor, VEGF)、组胺(histamine, HIS)浓度。生理盐水组予术后6h、24h、48h、72h静脉注射异硫氰酸罗丹明B标记的牛血清白蛋白(RBITC-BSA),注射后2min在激光共聚焦显微镜下观察兔肠系膜微血管白蛋白的分布、渗漏情况,并计算微血管白蛋白渗透率；羟乙基淀粉组组予术后6h、24h、48h、72h先静脉注射异硫氰酸荧光素标记的羟乙基淀粉(FITC-HES),注射后2min在激光共聚焦显微镜下观察兔肠系膜微血管内羟乙基淀粉的分布、渗漏情况,然后再注射异硫氰酸罗丹明B标记的牛血清白蛋白(RBITC-BSA),注射后2min在同一激光共聚焦显微镜视野下观察兔肠系膜微血管白蛋白的分布、渗漏情况,并计算微血管白蛋白渗透率；透射电镜观察术前对照组、生理盐水组术后48h和羟乙基淀粉组术后48h之间兔肠系膜微血管内皮细胞间隙的差异。结果：1、血清白蛋白浓度变化：①生理盐水组各组组内比较：术后6h、24h、48h、72h与术前相比均降低,其差异有统计学意义(P0.01),其中术后48h降低最明显。②羟乙基淀粉组各组组内比较：术后6h、24h、48h、72h与术前相比均降低,其差异有统计学意义(P0.01),其中术后48h降低最明显。③两组组间比较：术后6h、24h、48h、72h生理盐水组与羟乙基淀粉组相比差异均有统计学意义(P0.01)。2、血清炎性介质浓度比较：(1)血清IL-6：①生理盐水组各组组内比较：术后6h、24h、48h、72h与术前相比差异均有统计学意义(P0.01)。②羟乙基淀粉组各组组内比较：术后6h、24h、48h、72h与术前相比差异均有统计学意义(P0.01)。③两组组间比较：术后6h、24h、48h、72h生理盐水组与羟乙基淀粉组相比差异均有统计学意义(P0.01)。(2)血清TNF-α:①生理盐水组各组组内比较：术后6h、24h、48h、72h与术前相比差异均有统计学意义(P0.01)。②羟乙基淀粉组各组组内比较：术后6h与术前相比差异无统计学意义(P0.05),24h、48h、72h与术前相比差异均有统计学意义(P0.01)。③两组组间比较：术后6h、24h、48h、72h生理盐水组与羟乙基淀粉组相比差异均有统计学意义(P0.01)。(3)血清IL-2：①生理盐水组各组组内比较：术后6h、24h、48h、72h与术前相比差异均有统计学意义(P0.01)。②羟乙基淀粉组各组组内比较：术后6h、24h、48h、72h与术前相比差异均有统计学意义(P0.01)。③两组组间比较：术后6h、24h生理盐水组与羟乙基淀粉组相比差异均有统计学意义(P0.01)。术后72h生理盐水组与羟乙基淀粉组相比差异有统计学意义(P0.05)。术后48h生理盐水组与羟乙基淀粉组相比差异无统计学意义(P0.05)。(4)血清ICAM-1:①生理盐水组各组组内比较：术后6h、24h、48h、72h与术前相比差异均有统计学意义(P0.01)。②羟乙基淀粉组各组组内比较：术后6h、24h、48h、72h与术前相比差异均有统计学意义(P0.01)。③两组组间比较：术后6h、24h、48h、72h生理盐水组与羟乙基淀粉组相比差异均有统计学意义(P0.01)。(5)血清VEGF:①生理盐水组各组组内比较：术后6h、24h、48h、72h与术前相比差异均有统计学意义(P0.01)。②羟乙基淀粉组各组组内比较：术后6h与术前相比差异无统计学意义(P0.05),术后24h、48h、72h与术前相比差异均有统计学意义(P0.01)。③两组组间比较：术后6h、24h、48h、72h生理盐水组与羟乙基淀粉组相比差异均有统计学意义(P0.01)。(6)血清HIS：①生理盐水组各组组内比较：术后6h、24h、48h、72h与术前相比差异均有统计学意义(P0.01)。②羟乙基淀粉组各组组内比较：术后6h、24h、48h、72h与术前相比差异均有统计学意义(P0.01)。③两组组间比较：术后6h、24h、48h生理盐水组与羟乙基淀粉组相比差异均有统计学意义(P0.01)。术后72h生理盐水组与羟乙基淀粉组相比有差异有统计学意义(P0.05)。3、兔肠系膜微血管渗漏率的比较：①生理盐水组白蛋白渗漏率和羟乙基淀粉组白蛋白渗漏率比较,术后6h、24h、48h、72h渗漏率相比差异均有统计学意义(p0.01);②羟乙基淀粉组羟乙基淀粉渗漏率和白蛋白渗漏率比较,术后6h、24h、48h、72h渗漏率相比差异均有统计学意义(pO.01)。4、肠系膜微血管内皮细胞间隙变化：在术前对照组中,兔肠系膜微血管内皮细胞胞膜完整,连接紧密；在羟乙基淀粉组中,兔肠系膜微血管内皮细胞胞膜较完整,连接较紧密,内皮细胞间形成约0.1 μ m裂隙；在生理盐水组中,兔肠系膜微血管内皮细胞肿胀明显,内皮细胞间形成0.2-0.3 μ m裂隙。结论：1、兔腹部大手术后早期存在血清白蛋白浓度降低,术后48h血清白蛋白浓度降至最低；使用羟乙基淀粉(130/0.4)干预能减轻手术创伤后血浆白蛋白浓度下降幅度。2、兔腹部大手术后早期可出现血清炎性介质(IL-6、TNF-α、IL-2、ICAM-1、VEGF、HIS)浓度普遍升高；术后使用羟乙基淀粉干预可以降低IL-6、TNF-α、 IL-2、ICAM-1、VEGF的浓度,但不能抑制组胺(HIS)浓度升高。3、激光共聚焦显微镜观察到兔腹部大手术后早期微血管白蛋白和羟乙基淀粉渗漏,且二者都以术后48h渗漏最明显,羟乙基淀粉渗漏率较白蛋白低,术后使用羟乙基淀粉干预可以减轻肠系膜微血管白蛋白渗漏。4.电镜下可见术后早期微血管内皮细胞间隙异常开放。使用羟乙基淀粉干预可以减少微血管内皮细胞间隙的异常开放。
[Abstract]:Objective: To observe the early mesenteric microvascular albumin leakage, the intervention of hydroxyethyl starch, and the changes in the mesenteric microvascular endothelial cell space, and to determine the leakage of microvascular albumin and hydroxyethyl starch in mesenteric microvessels, the concentration of serum albumin and the concentration of serum inflammatory mediators. The effect of basic starch on the early microvascular albumin leakage after the major abdominal operation in rabbits. Methods: 54 Japanese big ear rabbits were randomly divided into 6 rats in the pre operation control group, 24 rats in the saline group after operation (6h, 24h, 48h, 72h each 6) and 24 hydroxyethyl starch group after operation (6h, 24h, 48h, 72h each 6). The two operation group all underwent gastrectomy plus the tail of pancreas, spleen, spleen and spleen. All rabbits were treated with heat card per kilogram, equal nitrogen nutrition support, heat 60kca/Kg, supplied by glucose and fat, provided 70% of the heat, 30%. nitrogen source was supplied by compound amino acids, the nitrogen heat ratio was 1:130., and 25ml/Kg was given to the physiological saline group to the observation time per day after the operation. The powder group was given 6%HES (130,0.4) 25ml/kg to the observation time every day, and the control group (no operation) was used as the detection point, the saline group and the hydroxyethyl starch group were used as the detection point after the operation (6h, 24h, 48h, 72h), and the serum albumin concentration was detected at every detection point, serum interleukin -6 (Interleukin-6, IL-6), tumor necrosis factor - alpha (T). Umour necrosis factor- alpha, TNF- alpha), interleukin -2 (Interleukin-2, IL-2), intercellular adhesion molecule -1 (intercellular adhesion molecule-1, ICAM-1), vascular endothelial growth factor, histamine concentration. Luo Danming B labeled bovine serum albumin (RBITC-BSA), the distribution, leakage, and permeability of microvascular albumin in rabbit mesenteric microvessel were observed under the laser confocal microscope after the injection of 2min, and the hydroxyethyl starch group was given 6h, 24h, 48h, and 72h first intravenous injection of hydroxyethyl starch labeled with fluorescein isothiocyanate (FITC) in the group of hydroxyethyl starch (FITC). -HES), after the injection of 2min, the distribution of hydroxyethyl starch in the rabbit mesenteric microvessel was observed under the laser confocal microscope, and the leakage was observed and then the bovine serum albumin (RBITC-BSA) was injected with isothiocyanate Luo Danming B. The distribution of the rabbit mesenteric microvascular albumin was observed under the same laser confocal microscope, and the distribution of the microvascular albumin in the rabbit mesentery was observed. Leakage, and calculation of microvascular albumin permeability; transmission electron microscopy to observe the differences in the mesenteric microvascular endothelial cell gap between the 48h and the hydroxyethyl starch group after the operation of the saline group. Results: 1, the change of serum albumin concentration: (1) the comparison of the serum albumin concentration in the saline group: the postoperative 6h, 24h, 48h, 72h and the operation in the saline group The difference was statistically significant (P0.01), and the postoperative 48h decreased most obviously. 2. In the group of hydroxyethyl starch groups, 6h, 24h, 48h, 72h were lower after operation than before operation, and the difference was statistically significant (P0.01), and the postoperative 48h decreased most obviously. (3) the two groups were compared: postoperative 6h, 24h, 48h, 72h physiological saline group. Compared with the hydroxyethyl starch group, the difference was statistically significant (P0.01).2, and the serum levels of inflammatory mediators were compared: (1) serum IL-6: (1) the comparison of 6h, 24h, 48h, 72h in the group after operation was statistically significant (P0.01). (2) the comparison of the groups in the group of hydroxyethyl starch: postoperative 6h, 24h, 48h, 72h and preoperative The difference was statistically significant (P0.01). (3) the comparison between the two groups: postoperative 6h, 24h, 48h, 72h physiological saline group compared with the hydroxyethyl starch group, the difference was statistically significant (P0.01). (2) serum TNF- A: (1) the physiological saline group group comparison: postoperative 6h, 24h, 48h, 72h compared with preoperative statistical significance (P0.01). (2) hydroxyethyl starch Group comparison: there was no significant difference in 6h after operation (P0.05), 24h, 48h, 72h and preoperative differences were statistically significant (P0.01). (3) the two groups were compared: postoperative 6h, 24h, 48h, 72h saline group compared with the hydroxyethyl starch group was statistically significant (P0.01). (3) serum IL-2: 1 physiological saline group, each Comparison in group group: 6h, 24h, 48h, 72h after operation were statistically significant compared with preoperative (P0.01). (2) the comparison of 6h, 24h, 48h, 72h (P0.01) in postoperative 6h, 24h, 48h, 72h compared with preoperative (P0.01). (3) the comparison of postoperative 6H, 24h physiological saline group and hydroxyethyl starch group had statistical difference. Significance (P0.01). Compared with the hydroxyethyl starch group, there was significant difference between the 72h saline group and the hydroxyethyl starch group (P0.05). There was no significant difference between the 48h saline group and the hydroxyethyl starch group after the operation (P0.05). (4) the comparison of the serum ICAM-1: (ICAM-1:) group in the saline group: 6h, 24h, 48h and 72h were statistically significant (P) after operation. 0.01) (0.01) 2 groups in the hydroxyethyl starch group: after the operation, the difference in 6h, 24h, 48h and 72h was statistically significant (P0.01). (3) the comparison between the two groups: postoperative 6h, 24h, 48h, 72h physiological saline group compared with the hydroxyethyl starch group was statistically significant (P0.01). (5) the comparison of serum VEGF: (1) group of saline group: 6 after operation H, 24h, 48h, 72h were statistically significant compared with preoperative (P0.01). 2. There was no significant difference in 6h from preoperative (P0.05), 24h, 48h, 72h were statistically significant (P0.01) compared with preoperative (P0.01). (3) the two groups were compared: postoperative 6h, saline, and hydroxyethyl The difference in starch group was statistically significant (P0.01). (6) serum HIS: (1) the comparison of 6h, 24h, 48h, 72h in each group after operation was statistically significant (P0.01). (2) the comparison of the groups in the group of hydroxyethyl starch: 6h, 24h, 48h, 72h compared with preoperative (P0.01). (3) two groups Comparison: the difference of 6h, 24h, 48h saline group and hydroxyethyl starch group after operation was statistically significant (P0.01). The difference between 72h physiological saline group and hydroxyethyl starch group after operation was statistically significant (P0.05).3, the comparison of the leakage rate of rabbit mesenteric microvascular: (1) the albumin leakage rate in the saline group and the hydroxyethyl starch group white egg Compared with the white leakage rate, the postoperative 6h, 24h, 48h, 72h leakage rates were statistically significant (P0.01). (2) compared with the hydroxyethyl starch leakage rate and the albumin leakage rate in the hydroxyethyl starch group, the postoperative 6h, 24h, 48h, and 72h leakage rates were statistically significant (pO.01).4, the mesenteric microvascular endothelial cell gap changes: in the pre operation control group In the group of hydroxyethyl starch, the membrane of the rabbit mesenteric microvascular endothelial cells was more complete and closely connected, and the endothelial cells formed about 0.1 M fissure in the hydroxyethyl starch group. In the saline group, the rabbit mesenteric microvascular endothelial cells were swollen obviously and the endothelial cells formed 0.2-0.3 mu m. Conclusions: 1. 1, the serum albumin concentration decreased in the early stage after the major abdominal operation and the concentration of serum albumin decreased to the lowest level after operation. The intervention of hydroxyethyl starch (130/0.4) could reduce the decrease of plasma albumin concentration of.2 after the operation, and the serum inflammatory mediators (IL-6, TNF- a, IL-2, ICAM-1) could be found at the early stage of the major abdominal operation in rabbits. The concentration of VEGF, HIS) was generally elevated, and the concentration of IL-6, TNF- a, IL-2, ICAM-1, VEGF could be reduced after the intervention of hydroxyethyl starch, but the concentration of histamine (HIS) could not be inhibited and.3 was not inhibited. The early microvascular albumin and hydroxyethyl starch leakage was observed by laser confocal microscope after the major operation of the rabbit abdomen, and the two were most obvious in the postoperative 48h leakage. The leakage rate of hydroxyethyl starch is lower than that of albumin. The intervention of hydroxyethyl starch after operation can reduce the microvascular albumin leakage in mesenteric microvessel.4. under the electron microscope, and it can be seen that the early microvascular endothelial gap in the microvascular spaces is abnormal opening. The intervention of hydroxyethyl starch can reduce the abnormal opening of the intercellular space of microvascular endothelial cells.

【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R656

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