围术期利多卡因对宫颈癌根治术患者淋巴细胞功能及HMGB1释放的影响

发布时间：2018-01-14 14:43

本文关键词：围术期利多卡因对宫颈癌根治术患者淋巴细胞功能及HMGB1释放的影响　出处：《山东大学》2014年硕士论文　论文类型：学位论文

【摘要】：研究背景及目的：外科手术及创伤可使机体内分泌系统的下丘脑-垂体-肾上腺轴(HPA)功能增强,增强交感神经系统的应激反应及急性期的应答反应,引发机体应激性炎症反应甚至是全身性炎症反应综合症,这一过程伴随着机体免疫系统的激活,促炎介质呈高水平加工及释放,最终导致机体免疫功能迅速受抑,而手术应激导致的免疫抑制效应,是造成术后并发感染及加速残存肿瘤转移的重要原因。利多卡因作为一种常用的酰胺类局部麻醉药,不仅具有镇痛、抗痛觉过敏、抗心律失常的作用,研究显示其还具有抗炎作用,且围术期静脉输注利多卡因能够减轻术后疼痛,减少阿片类镇痛药使用量,减轻机体炎症反应,加快胃肠功能恢复及缩短患者住院时间。高迁移率族蛋白B1(HMGB1)在多种恶性肿瘤中表达异常增高,作为一种晚期炎症性蛋白,已有研究显示其可介导机体对感染及损伤的炎症反应,从而促进炎症反应的发生及发展。本研究通过观察术中持续静脉输注小剂量利多卡因对淋巴细胞增殖、凋亡、Th1/Th2漂移及HMGB1释放的影响,探讨利多卡因对手术创伤致机体免疫功能下降的保护作用及机制,为利多卡因应用于临床抗炎治疗提供理论依据。研究方法：选择2013年6月至2014年1月已确诊宫颈癌且于山东大学齐鲁医院第一手术室择期行全身麻醉下宫颈癌根治术(广泛子宫切除术加盆腔淋巴结清扫术)患者30例,所有患者都签署医学试验知情同意书,采用随机数字表法,将病人随机分为对照组(C)和利多卡因组(L),每组各15例(n=15),两组病人临床基线无统计学差异。气管插管静吸复合麻醉。病人进入手术间后开放利多卡因专用静脉通道,麻醉诱导前15min静推利多卡因1.5mg/kg(推注时间持续约10min),麻醉诱导后微量泵静脉输注利多卡因1.5mg·kg-1·h-1至病人离开手术间。对照组给予相等容量的生理盐水。手术前24h、手术后Oh、手术后48h分别抽取病人静脉血10ml。离心,收集血浆,分离外周血淋巴细胞。ELISA法测定各时间点血浆HMGB1、IFN-γ及IL-4浓度,通过IFN-γ与IL-4浓度的比值反映Th1/Th2细胞因子平衡的情况,评估两组机体免疫功能的变化差异,通过HMGB1浓度值比较手术前后两组HMGB1浓度的变化,CCK-8法进行PHA诱导的淋巴细胞增殖试验,流式Annexin V-PI双染法进行淋巴细胞凋亡检测。研究结果： 1、两组病人年龄、体重等无统计学差异(p0.05)； 2、相比于术前24h,与C组比较,L组术后oh(0.448±0.139vs.0.352±0.112OD)及术后48h(0.420±0.094vs.0.326±0.141OD) PHA诱导的淋巴细胞增殖试验中淋巴细胞活性下降减少(p0.05)； 3、流式Annexin V-PI双染法进行淋巴细胞凋亡检测,两组淋巴细胞凋亡比例在术后都有所增加,与C组相比,L组术后48h淋巴细胞凋亡上升幅度较低(3.560±0.775vs.4.173±0.779%),差异有统计学意义(p0.05)； 4、与C组比较,L组术后48hIFN-y的浓度(4.041±0.230vs.3.782±0.282pg/ml)升高,差异有统计学意义(p0.05)；分别计算两组IFN-y/IL-4的比值,与C组比较,L组比值升高(0.676±0.288vs.0.489±0.124,p0.05)； 5、与C组比较,L组术后48h的HMGB1表达浓度下降更明显(53.458±8.983vs.59.387±5.025μg/L),差异有统计学意义(p0.05)。研究结论： 1、围术期静脉输注利多卡因可减轻手术创伤对淋巴细胞增殖能力的抑制作用； 2、围术期静脉输注利多卡因可减少淋巴细胞术后凋亡； 3、围术期静脉输注利多卡因可抑制机体免疫调节平衡向Th2漂移,保护机体免疫功能； 4、围术期静脉输注利多卡因对机体免疫功能的调节作用可能与其能下调高迁移率族蛋白B1(HMGB1)的表达有关。研究意义：本研究通过观察静脉输注利多卡因对机体免疫系统的影响,一方面验证了利多卡因的抗炎作用,为临床应用利多卡因提供理论依据：一方面提出了利多卡因对机体免疫功能的保护作用可能是通过其抑制晚期炎症因子高迁移率族蛋白B1(HMGB1)来实现的假设,为临床抗炎治疗提供了新思路,但这一假设的确切机制尚未明确,还待进一步的深入研究。
[Abstract]:Research background and purpose:
Surgical trauma and the endocrine system of the hypothalamic pituitary adrenal axis (HPA) function enhancement, enhanced responses to stress response of the sympathetic nervous system and acute stress, triggering the body's inflammatory response and systemic inflammatory response syndrome, this process is accompanied by the activation of the immune system, proinflammatory mediators a processing and release of high levels, resulting in rapid inhibition of immune function, and immune suppression caused the effect of surgical stress, is an important cause of infection and accelerate the transfer of residual tumor after surgery. Lidocaine is a kind of common amide local anesthetics, not only has analgesic, anti hyperalgesia, anti arrhythmia research shows that the effect also has anti-inflammatory effect, and perioperative intravenous infusion of lidocaine can reduce postoperative pain, reduce opioid analgesic usage, relieve inflammation In response, accelerate the recovery of gastrointestinal function and shorten the hospitalization time. High mobility group protein B1 (HMGB1) abnormal expression in a variety of malignant tumors, as a late inflammatory protein, has been shown to mediate the inflammatory response to infection and injury, so as to promote the occurrence and development of inflammatory reaction this study. Through continuous intravenous injection of small dose of lidocaine on apoptosis of lymphocyte proliferation were observed, Th1/Th2 drift effects and released by HMGB1, this study was designed to investigate the protective effect and mechanism of immunity of surgical trauma, and provide a theoretical basis for the clinical application of lidocaine anti-inflammatory treatment.
Research methods:
From June 2013 to January 2014 has been diagnosed with cervical cancer in Qilu Hospital of Shandong University and the first operation room undergoing radical hysterectomy under general anesthesia (hysterectomy and pelvic lymphadenectomy) in 30 patients, all patients signed informed consent to medical tests, using the random number table method, the patients were randomly divided into control group (C) and lidocaine group (L), 15 cases in each group (n=15), no significant difference between the two groups of patients with clinical baseline. Tracheal intubation anesthesia. The patient into the operating room after open special lidocaine intravenous access, 15min before induction of anesthesia (intravenous lidocaine 1.5mg/kg injection duration is about 10min), 1.5mg injection of lidocaine anesthesia kg-1 H-1 to the patients with micro pump intravenous induction after leaving the operating room. The control group received equal volume normal saline. 24h before surgery, Oh after surgery, 48h after surgery were selected with static 10ml. vein blood centrifugation, plasma was collected, separation of peripheral blood lymphocyte.ELISA method for determination of plasma HMGB1 at each time point, IFN- y and IL-4 concentration, reflecting Th1/Th2 cytokine balance by the ratio of IFN- gamma and IL-4 concentration changes, assessment of two groups of immune function, the value changes compared before and after the operation of two groups of HMGB1 concentration through the method of CCK-8 HMGB1 concentration, PHA induced lymphocyte proliferation test, lymphocyte apoptosis were detected by flow cytometry with Annexin V-PI double staining method.
The results of the study:
1, there was no statistical difference in age and weight between the two groups (P0.05).
2, compared with preoperative 24h, compared with group C, the activity of lymphocytes in group L decreased (P0.05) after Oh (0.448 + 0.139vs.0.352 + 0.112OD) and 48h (0.420 + 0.094vs.0.326 + 0.141OD) PHA induced lymphocyte proliferation test.
3, the apoptosis rate of lymphocytes was detected by flow Annexin V-PI double staining. The proportion of lymphocyte apoptosis increased in two groups after operation. Compared with C group, the apoptosis rate of 48h lymphocyte in L group increased significantly (3.560 + 0.775vs.4.173 + 0.779%), the difference was statistically significant (P0.05).
4, compared with group C, the concentration of 48hIFN-y in group L increased (4.041 + 0.230vs.3.782 + 0.282pg/ml), and the difference was statistically significant (P0.05). The ratio of IFN-y/IL-4 in the two groups was calculated. Compared with C group, the ratio of L group increased (0.676 + 0.288vs.0.489 + 0.124, P0.05).
5, compared with the C group, the HMGB1 expression concentration of 48h in group L decreased significantly (53.458 + 8.983vs.59.387 + 5.025 g/L), and the difference was statistically significant (P0.05).
The conclusions are as follows:
1, intravenous infusion of lidocaine during perioperative period can reduce the inhibitory effect of surgical trauma on the proliferation of lymphocyte.
2, intravenous infusion of lidocaine during perioperative period can reduce apoptosis after lymphocyte operation.
3, intravenous infusion of lidocaine during perioperative period can inhibit the shift of immunoregulation balance to Th2 and protect the immune function of the body.
4, the regulation of intravenous infusion of lidocaine on the immune function of the body may be related to the ability to reduce the expression of high mobility group protein B1 (HMGB1).
Research significance:
This study observed the effects of intravenous infusion of lidocaine on the immune system, verify the anti-inflammatory effect of lidocaine on the one hand, to provide a theoretical basis for the clinical application of lidocaine: on the one hand, put forward the protective effect of lidocaine on immune function may be through the inhibition of late inflammatory factor of high mobility group protein B1 (HMGB1) to achieve that provides a new idea for clinical anti-inflammatory treatment, but the exact mechanism of this hypothesis is not yet clear, still needs further research.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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