针刀医学内脏弓弦原理指导下韧带机械损伤致兔CPID模型的建立

发布时间：2018-01-26 15:26

本文关键词： 针刀内脏弓弦慢性盆腔炎模型　出处：《湖北中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：实验目的:以针刀医学内脏弓弦原理对慢性盆腔炎的认识为理论依据,采用机械损伤新西兰兔子宫阔韧带的方法建立慢性盆腔炎的模型,旨在探索一种新的理论指导下慢性盆腔炎动物模型的造模方法,为针刀医学内脏弓弦理论及针刀临床治疗慢性内脏疾病提供实验学基础。实验方法:将普通级康健雌性未孕新西兰大白兔28只适应性饲养一周,随机分为空白对照组、假手术组、造模组和机械损伤加混合菌对照组,每组各7只。其中,空白对照组不采取任何干预措施,正常饲养;其余三组在造模前一天禁食、禁水12h,10%水合氯醛(3ml/kg)耳缘静脉麻醉后备皮,在严格遵守无菌手术条件下开腹,假手术组不作任何处理,分层关腹;造模组找到子宫阔韧带用刀柄刮划2-3下、局部折叠缝合使子宫失去正常位置后分层关腹;机械损伤加混合菌对照组用注射器抽插损伤子宫内膜后,在子宫角向卵巢方向呈锐角向两侧输卵管各注入0.15ml混合菌液(大肠埃希氏菌、金黄色葡萄球菌、乙型溶血性链球菌按2:1:1比例用无菌生理盐水稀释,配成3×109个/ml浓度)。术后常规饲养,按实验设定日期空白对照组、假手术组和机械损伤加混合菌对照组21天取材,造模组60天取材。肉眼观察子宫及盆腔组织形态学变化后,取子宫组织HE染色,在光镜下观察其病理改变;心脏取血2ml置于肝素抗凝管中,用酶联免疫法测各组兔血清的炎症细胞因子IL-1β、IL-6、TNF-α水平。实验结果:(1)肉眼观察:造模组可见子宫表面浅表血管扩张、色鲜红,子宫增粗、肿胀迂曲或积水,质脆、弹性差,与周围脏器、盆腔组织有不同程度的纤维组织粘连,不易分离,或有串珠状改变,内可见淡黄色液体。(2)光镜下:造模组子宫黏膜内的上皮细胞遭到破坏,构造失常、层次模糊不清;黏膜层、肌层有慢性炎性细胞浸润及纤维结缔组织增生,以淋巴细胞为主;腺体破坏甚至坏死。(3)指标检测结果:造模组兔血清酶联免疫吸附法测得IL-1β、IL-6、TNF-α值均较空白对照组和假手术组升高,差异有统计学意义(P0.05);与机械损伤加混合菌对照组比,差异无统计学意义(P0.05)。实验结论:根据针刀医学内脏弓弦力学解剖系统理论对慢性盆腔炎的认识,采用人为机械损伤子宫阔韧带、局部折叠缝合使子宫失去正常位置的方法初步探索慢性盆腔炎动物模型,其形态学观察有病理改变、血清指标有炎性变化,但需进一步减少人为干预、完善检测指标造模方法仍须进一步完善。
[Abstract]:Objective: to establish a model of chronic pelvic inflammatory disease by mechanical injury of the broad uterine ligament in New Zealand rabbits based on the understanding of chronic pelvic inflammatory disease based on the principle of visceral bow string in needle knife medicine. The purpose of this study was to explore a new method for modeling chronic pelvic inflammatory disease (CPI) animal model under the guidance of new theory. Objective: to provide experimental basis for the treatment of chronic visceral diseases by acupuncture knife medical visceral bow string theory and clinical treatment of chronic visceral diseases. Methods: 28 healthy female New Zealand white rabbits were fed adaptively for one week. They were randomly divided into blank control group, sham operation group, model making group and mechanical injury plus mixed bacteria control group with 7 rats in each group. The other three groups fasted one day before the model and banned 10% chloral hydrate (10% chloral hydrate) 3 ml / kg for 12 hours. The patients in the sham operation group were treated without any treatment under strict compliance with aseptic operation. Stratified closed abdomen; In the model group, the broad ligament of uterus was scraped 2-3 times with the knife handle, and the uterus lost its normal position by local folding and suture, then the uterus was laminated and closed. Mechanical injury and mixed bacteria control group were injected with 0.15 ml mixed bacteria solution (Escherichia coli) in the direction of uterus horn to ovary and oviduct in the direction of acute angle after the injury of endometrium was inserted with syringe in the control group. Staphylococcus aureus and Streptococcus B hemolytic streptococcus were diluted with aseptic saline at the ratio of 2: 1: 1 to form 3 脳 10 9 / ml of saline. The sham-operation group and the mechanical injury plus mixed bacteria control group were taken for 21 days and the model group for 60 days. After observing the morphological changes of uterus and pelvic tissues with naked eyes, the uterine tissues were stained with HE. The pathological changes were observed under light microscope. 2ml blood was taken from the heart and placed in the heparin anticoagulant tube. The inflammatory cytokine IL-1 尾 -IL-6 was measured by enzyme-linked immunosorbent assay (Elisa). Experimental results: the model group showed superficial vasodilation, bright red color, thickening of uterus, swelling, detour or hydrops, brittle, poor elasticity, and surrounding organs. Pelvic tissue has different degrees of fibrous tissue adhesion, not easy to be separated, or beaded changes, the light yellow liquid can be seen in the light microscope: the model group uterine mucosa epithelial cells were destroyed, the structure is abnormal. The level is not clear; There were chronic inflammatory cell infiltration and fibrous connective tissue proliferation in mucous membrane and myometrium, mainly lymphocytes. Results: the serum IL-1 尾 -IL-6 TNF- 伪 values of the model group were higher than those of the control group and the sham-operation group, compared with the control group and the sham-operation group. The serum enzyme linked immunosorbent assay (Elisa) of the model group was higher than that of the blank control group and the sham-operated group. The difference was statistically significant (P 0.05). Compared with the control group of mechanical injury and mixed bacteria, there was no significant difference between the two groups (P0.055.Experimental conclusion: according to the theory of mechanical anatomy system of visceral bow string in needle knife medicine, the understanding of chronic pelvic inflammatory disease. The animal model of chronic pelvic inflammatory disease was preliminarily explored by artificial mechanical injury of the broad ligament of uterus and local folding and suture to make the uterus lose its normal position. There were pathological changes in morphology and inflammatory changes in serum indexes. However, it is necessary to further reduce human intervention and improve the method of modeling detection index.
【学位授予单位】：湖北中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R245;R-332

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