清热解毒方与凉血活血方防治术后腹腔粘连的实验研究
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摘要
一、术后腹腔粘连动物模型的建立与筛选
目的：比较研究8种大鼠术后腹腔粘连模型的特点及应用。方法：180只Wistar大鼠随机分为9组,每组20只,设假手术组和8个模型组,各组按不同手术方式制作模型。术后详细记录动物生存情况,存活大鼠于术后第7天处死剖验,按Nair五级评分标准对腹腔粘连进行评价,并取粘连组织送检病理。结果：E组(盲肠+切口两侧腹壁刷擦,100%)、D组(盲肠+切口右侧腹壁刷擦,90%)、Ⅰ组(止血钳钳夹十二指肠,75%)术后7天动物生存率明显高于其他各组。G组(盲肠结扎+盲端开口+滑石粉涂抹,3.000±0.926)、H组(盲肠结扎穿孔+盲肠刷擦,3.500±0.577)和Ⅰ组(止血钳钳夹十二指肠,3.270±0.704)粘连程度优于其他各组。结论：止血钳钳夹十二指肠致腹腔粘连模型是较理想的大鼠术后腹腔粘连模型。
二、十二指肠钳火致术后腹腔粘连大鼠模型的形态学研究
目的：观察术后14天内十二指肠钳夹致大鼠术后腹腔粘连形成过程中的组织学和形态学变化。方法：48只Wistar大鼠随机分为6组,分别在术后即刻、术后第1、3、5、7和14天处死,取手术或者粘连部位,应用光镜及扫描电镜进行组织学和形态学观察。结果：术后第1天时手术部位炎症最为严重,粘连形成的范围和强度分别在术后第3天和第5天时达到峰值,术后第7天时间皮细胞完全覆盖手术部位；肌成纤维细胞从术后第3天开始大量增殖。结论：十二指肠钳夹可以诱发明显的术后腹腔粘连,肌成纤维细胞在粘连形成过程中可能起重要作用。
三、清热解毒方与凉血活血方防治大鼠术后腹腔粘连的实验研究
目的：观察清热解毒方与凉血活轿方对大鼠术后腹腔粘连的防治效果并探索其机制。方法：160只Wistar大鼠随机分为假手术组、模型组、抗生索治疗组、瑞术康治疗组、清热解毒方组、凉血活血方组、两方合用组、两方序贯组等8组,制作十二指肠钳夹致大鼠术后腹腔粘连模型,并分别给药治疗7天。术后14天时处死剖验并对粘连情况评分,取手术部位及粘连组织分别进行HE染色和苦味酸-天狼星红染色,镜下观察并摄片,对粘连组织纤维化、炎症、血管增殖及Ⅰ、Ⅲ型胶原胶原含量进行评价。结果：4个中药治疗组生存情况与模型组无差异。清热解毒方组在术后腹腔粘连范围、强度评分低于其他治疗组。各组粘连主要涉及肝脏、十二指肠、大网膜及腹壁。清热解毒方组、凉血活血方组纤维化评分低于其他组,4个中药治疗组炎症评分相似,血管增殖评分以清热解毒方组为最低。清热解毒方组粘连组织中Ⅰ、Ⅲ型胶原含量低于其他组。结论：清热解毒方、凉血活血方、两方合用及两方序贯给药均有一定的抗粘连效果,其中以清热解毒方单用效果最佳；其抗粘连机理可能与其抗炎作用有关。
四、基于抗体芯片技术的术后腹腔感染大鼠细胞因子谱分析
目的：应用抗体芯片技术筛选术后腹腔感染大鼠特异性标志物。方法：20只Wistar大鼠随机分为实验组和假手术组并分别手术,术后3天内取大鼠腹腔灌洗液、血清、血浆及淋巴液送检,应用抗体芯片进行检测并筛选出差异表达蛋白。结果：从大鼠腹腔灌洗液、血清、血浆中分别筛选出19种、20种、22种差异表达蛋白。淋巴液检测未能得出结果。其中Integrin αMβ2、TRAIL两种细胞因子在3组标本中含量均有差异；CINC-2α/β、FADD、MuSK、IL-3、IL-4、 TIE-2、TLR4等7种细胞因子在3组标本中的2组表达有差异。结论：血浆和腹水是监测腹腔感染比较理想的标本,所筛选出的9种细胞因子的应用价值,需要更多证据支持。
1. Comparative study of8kinds of postoperative abdominal adhesion models
Objective:To study the features and application of8kinds of rat model of postoperative abdominal adhesion. Methods:180Wistar rats were randomly divided into9groups of20animals in each, including sham operation group and8model groups. The survival of each group was registered, survivors were sacrificed on the seventh post-operative day, and the degree of peritoneal adhesion was evaluated by Nair adhesion scale, and the adhesion tissues were examined by histopathology. Results:The survival rates of Group E, Group D and Group I were significantly higher than other model groups. The scores of adhesion of Group G, Group H and Group1were higher than other groups. Conclusion:Duodenum clamped using hemostat is an ideal method for postoperative abdominal adhesion rat model.
2. Duodenum clamping trauma induces significant postoperative intraperitoneal
Objective:To investigate the histological and morphological changes in the first14postoperative days on a rat intraperitoneal adhesions model induced by duodenum clamping trauma. Method:The rat model of postoperative intraperitoneal adhesions was established in48male Wistar rats by laparotomy, followed by the duodenum clamping trauma. Rats were sacrificed immediately after the operation (0day), and then on1st,3rd,5th,7th and14th day, respectively. The intraperitoneal adhesions were assessed macroscopically. Histopathology and immunohistochemistry were performed to evaluate the adhesion tissues. In addition, the changes of the mesothelium covering the surgical sites were examined by scanning electron microscopy. Results:The extent and tenacity of intraperitoneal adhesions reached their peaks on3rd and5th days, respectively. Histopathological examination showed that all rats developed inflammatory responses at the clamped sites of duodenum, which was most prominent on1st day. The scores of fibrosis and vascular proliferation increased slowly from3rd to5th day. Myofibroblasts proliferated significantly in the adhesion tissues from3rd day, which were examined by immunohistochemical method. And the mesothelium covering the surgical sites and the adhesion tissues healed on7th day. Conclusion:The clamping trauma to the duodenum can result in significant postoperative intraperitoneal adhesions formation, which represents an ideal rat model for intraperitoneal adhesions research and prevention. And myofibroblasts may play an important role in the forming process of intraperitoneal adhesions.
3. Experiment study of the prevention of QingreJiedu Herbs and LiangxueHuoxue Herbs on postoperative abdominal adhesion in rats
Objective:To observe the prevention of QingreJiedu Herbs and LiangxueHuoxue Herbs on postoperative abdominal adhesion in rats and explore the mechanism. Method:160Wistar rats were randomly divided into8groups and then made into postoperative abdominal adhesion model by duodenum clamped; protective measures or medicines were given respectively in the7days. Adhesions war evaluated on the14th post-surgical day, specimens were collected for histological staining and grading. Results:QingreJiedu Herbs, LiangxueHuoxue Herbs and their combinations can reduce the adhesion formation after the operation. QingreJiedu Herbs can inhibit the inflammation, the fibrosis and the neovasculrization of the surgical sites or adhesion tissues, and the formation and deposition of type Ⅰ, Ⅲ collagen were also depressed. Conclusion:QingreJiedu Herbs, LiangxueHuoxue Herbs, their combining and sequential applications can all depress the postoperative abdominal adhesion formation; the effect of QingreJiedu Herbs is the best. And the anti-adhesion mechanism may be related to their anti-inflammatory effects.
4. An antibody microarray assay of postoperative abdominal infection in rats
Objective:To screen specific markers of postoperative abdominal infection in rats using an antibody microarray. Method:20Wistar rats were randomly divided into experimental group and control group; peritoneal ravage fluid, serum, plasma and lymph fluid were collected3days after the operation, and tested using antibody chips. Results:19,20and22kinds of proteins were respectively elected from the peritoneal lavage fluid, serum and plasma. Lymphatic fluid failed to result. Among which, Integrity αMβ2, TRAIL singled out in3kinds of samples, and CINC-2α/β, FADD, MuSK, IL-3, IL-4, TIE-2, TLR4in2. Conclusion:The serum and peritoneal lavage fluid are ideal specimens for monitoring postoperative abdominal infection, the value of the9cytokines needs more supporting information.
目的：比较研究8种大鼠术后腹腔粘连模型的特点及应用。方法：180只Wistar大鼠随机分为9组,每组20只,设假手术组和8个模型组,各组按不同手术方式制作模型。术后详细记录动物生存情况,存活大鼠于术后第7天处死剖验,按Nair五级评分标准对腹腔粘连进行评价,并取粘连组织送检病理。结果：E组(盲肠+切口两侧腹壁刷擦,100%)、D组(盲肠+切口右侧腹壁刷擦,90%)、Ⅰ组(止血钳钳夹十二指肠,75%)术后7天动物生存率明显高于其他各组。G组(盲肠结扎+盲端开口+滑石粉涂抹,3.000±0.926)、H组(盲肠结扎穿孔+盲肠刷擦,3.500±0.577)和Ⅰ组(止血钳钳夹十二指肠,3.270±0.704)粘连程度优于其他各组。结论：止血钳钳夹十二指肠致腹腔粘连模型是较理想的大鼠术后腹腔粘连模型。
二、十二指肠钳火致术后腹腔粘连大鼠模型的形态学研究
目的：观察术后14天内十二指肠钳夹致大鼠术后腹腔粘连形成过程中的组织学和形态学变化。方法：48只Wistar大鼠随机分为6组,分别在术后即刻、术后第1、3、5、7和14天处死,取手术或者粘连部位,应用光镜及扫描电镜进行组织学和形态学观察。结果：术后第1天时手术部位炎症最为严重,粘连形成的范围和强度分别在术后第3天和第5天时达到峰值,术后第7天时间皮细胞完全覆盖手术部位；肌成纤维细胞从术后第3天开始大量增殖。结论：十二指肠钳夹可以诱发明显的术后腹腔粘连,肌成纤维细胞在粘连形成过程中可能起重要作用。
三、清热解毒方与凉血活血方防治大鼠术后腹腔粘连的实验研究
目的：观察清热解毒方与凉血活轿方对大鼠术后腹腔粘连的防治效果并探索其机制。方法：160只Wistar大鼠随机分为假手术组、模型组、抗生索治疗组、瑞术康治疗组、清热解毒方组、凉血活血方组、两方合用组、两方序贯组等8组,制作十二指肠钳夹致大鼠术后腹腔粘连模型,并分别给药治疗7天。术后14天时处死剖验并对粘连情况评分,取手术部位及粘连组织分别进行HE染色和苦味酸-天狼星红染色,镜下观察并摄片,对粘连组织纤维化、炎症、血管增殖及Ⅰ、Ⅲ型胶原胶原含量进行评价。结果：4个中药治疗组生存情况与模型组无差异。清热解毒方组在术后腹腔粘连范围、强度评分低于其他治疗组。各组粘连主要涉及肝脏、十二指肠、大网膜及腹壁。清热解毒方组、凉血活血方组纤维化评分低于其他组,4个中药治疗组炎症评分相似,血管增殖评分以清热解毒方组为最低。清热解毒方组粘连组织中Ⅰ、Ⅲ型胶原含量低于其他组。结论：清热解毒方、凉血活血方、两方合用及两方序贯给药均有一定的抗粘连效果,其中以清热解毒方单用效果最佳；其抗粘连机理可能与其抗炎作用有关。
四、基于抗体芯片技术的术后腹腔感染大鼠细胞因子谱分析
目的：应用抗体芯片技术筛选术后腹腔感染大鼠特异性标志物。方法：20只Wistar大鼠随机分为实验组和假手术组并分别手术,术后3天内取大鼠腹腔灌洗液、血清、血浆及淋巴液送检,应用抗体芯片进行检测并筛选出差异表达蛋白。结果：从大鼠腹腔灌洗液、血清、血浆中分别筛选出19种、20种、22种差异表达蛋白。淋巴液检测未能得出结果。其中Integrin αMβ2、TRAIL两种细胞因子在3组标本中含量均有差异；CINC-2α/β、FADD、MuSK、IL-3、IL-4、 TIE-2、TLR4等7种细胞因子在3组标本中的2组表达有差异。结论：血浆和腹水是监测腹腔感染比较理想的标本,所筛选出的9种细胞因子的应用价值,需要更多证据支持。
1. Comparative study of8kinds of postoperative abdominal adhesion models
Objective:To study the features and application of8kinds of rat model of postoperative abdominal adhesion. Methods:180Wistar rats were randomly divided into9groups of20animals in each, including sham operation group and8model groups. The survival of each group was registered, survivors were sacrificed on the seventh post-operative day, and the degree of peritoneal adhesion was evaluated by Nair adhesion scale, and the adhesion tissues were examined by histopathology. Results:The survival rates of Group E, Group D and Group I were significantly higher than other model groups. The scores of adhesion of Group G, Group H and Group1were higher than other groups. Conclusion:Duodenum clamped using hemostat is an ideal method for postoperative abdominal adhesion rat model.
2. Duodenum clamping trauma induces significant postoperative intraperitoneal
Objective:To investigate the histological and morphological changes in the first14postoperative days on a rat intraperitoneal adhesions model induced by duodenum clamping trauma. Method:The rat model of postoperative intraperitoneal adhesions was established in48male Wistar rats by laparotomy, followed by the duodenum clamping trauma. Rats were sacrificed immediately after the operation (0day), and then on1st,3rd,5th,7th and14th day, respectively. The intraperitoneal adhesions were assessed macroscopically. Histopathology and immunohistochemistry were performed to evaluate the adhesion tissues. In addition, the changes of the mesothelium covering the surgical sites were examined by scanning electron microscopy. Results:The extent and tenacity of intraperitoneal adhesions reached their peaks on3rd and5th days, respectively. Histopathological examination showed that all rats developed inflammatory responses at the clamped sites of duodenum, which was most prominent on1st day. The scores of fibrosis and vascular proliferation increased slowly from3rd to5th day. Myofibroblasts proliferated significantly in the adhesion tissues from3rd day, which were examined by immunohistochemical method. And the mesothelium covering the surgical sites and the adhesion tissues healed on7th day. Conclusion:The clamping trauma to the duodenum can result in significant postoperative intraperitoneal adhesions formation, which represents an ideal rat model for intraperitoneal adhesions research and prevention. And myofibroblasts may play an important role in the forming process of intraperitoneal adhesions.
3. Experiment study of the prevention of QingreJiedu Herbs and LiangxueHuoxue Herbs on postoperative abdominal adhesion in rats
Objective:To observe the prevention of QingreJiedu Herbs and LiangxueHuoxue Herbs on postoperative abdominal adhesion in rats and explore the mechanism. Method:160Wistar rats were randomly divided into8groups and then made into postoperative abdominal adhesion model by duodenum clamped; protective measures or medicines were given respectively in the7days. Adhesions war evaluated on the14th post-surgical day, specimens were collected for histological staining and grading. Results:QingreJiedu Herbs, LiangxueHuoxue Herbs and their combinations can reduce the adhesion formation after the operation. QingreJiedu Herbs can inhibit the inflammation, the fibrosis and the neovasculrization of the surgical sites or adhesion tissues, and the formation and deposition of type Ⅰ, Ⅲ collagen were also depressed. Conclusion:QingreJiedu Herbs, LiangxueHuoxue Herbs, their combining and sequential applications can all depress the postoperative abdominal adhesion formation; the effect of QingreJiedu Herbs is the best. And the anti-adhesion mechanism may be related to their anti-inflammatory effects.
4. An antibody microarray assay of postoperative abdominal infection in rats
Objective:To screen specific markers of postoperative abdominal infection in rats using an antibody microarray. Method:20Wistar rats were randomly divided into experimental group and control group; peritoneal ravage fluid, serum, plasma and lymph fluid were collected3days after the operation, and tested using antibody chips. Results:19,20and22kinds of proteins were respectively elected from the peritoneal lavage fluid, serum and plasma. Lymphatic fluid failed to result. Among which, Integrity αMβ2, TRAIL singled out in3kinds of samples, and CINC-2α/β, FADD, MuSK, IL-3, IL-4, TIE-2, TLR4in2. Conclusion:The serum and peritoneal lavage fluid are ideal specimens for monitoring postoperative abdominal infection, the value of the9cytokines needs more supporting information.
引文
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