阅读说明：本技术 经腹膜后入路构建大鼠肾下腹主动脉瘤模型及构建方法 (Rat infrarenal abdominal aortic aneurysm model established through retroperitoneal approach and establishment method ) 是由 朱君星 杨颖� 唐全巧 石星驰 易思怡 周灿 冯松 于 2021-07-20 设计创作，主要内容包括：本发明涉及生物及医药技术的技术领域,特别是涉及经腹膜后入路构建大鼠肾下腹主动脉瘤模型及构建方法,操作相对简单,可避免经腹腔入路所致的风险,未采用加压灌注,缩短灌注时间至20min,无下肢缺血发生,且不影响成瘤率,对血管损伤极小,灌注结束后局部压迫便可止血,显著提高了手术成功率；包括以下步骤：S1、大鼠术前12h禁食,按40～50mg/kg,予戊巴比妥钠麻醉,麻醉满意后取右侧卧位,常规备皮,消毒,取左侧腋后线肋缘下至大腿根部间连线为切口,长3～3.5cm,逐层切开皮肤、皮下组织及肌肉,见腹膜后脂肪组织,并沿着脂肪组织找到腹膜与腰背肌潜在间隙,用棉签分离此间隙直达腹主动脉,分离腹主动脉与下腔静脉。(The invention relates to the technical field of biology and medicine technology, in particular to a rat infrarenal abdominal aortic aneurysm model constructed by retroperitoneal approach and a construction method, the operation is relatively simple, the risk caused by retroperitoneal approach can be avoided, pressurized perfusion is not adopted, the perfusion time is shortened to 20min, no lower limb ischemia occurs, the tumorigenicity rate is not influenced, the damage to blood vessels is extremely small, hemostasis can be realized by local compression after the perfusion is finished, and the success rate of the operation is obviously improved; the method comprises the following steps: s1, fasting the rat before operation for 12 hours, performing pentobarbital sodium anesthesia according to the ratio of 40-50 mg/kg, taking a right lateral lying position after satisfactory anesthesia, conventionally preparing skin, sterilizing, taking a connecting line from the left axillary posterior line, the costal margin to the thigh root as an incision, cutting skin, subcutaneous tissues and muscles by layers with the length of 3-3.5 cm, finding out fat tissues behind the peritoneum, finding out potential gaps between the peritoneum and the lumbodorsal muscles along the fat tissues, separating the gaps by using a cotton swab to directly reach the abdominal aorta, and separating the abdominal aorta from the inferior vena cava.)

1. The method for constructing the rat infrarenal abdominal aortic aneurysm model through retroperitoneal approach is characterized by comprising the following steps:

s1, fasting the rat before operation for 12 hours, performing pentobarbital sodium anesthesia according to 40-50 mg/kg, taking a right lateral lying position after satisfactory anesthesia, conventionally preparing skin, sterilizing, taking a connecting line from the left axillary posterior line, the costal margin to the thigh root as an incision, wherein the length is 3-3.5 cm, incising skin, subcutaneous tissues and muscles layer by layer to see fat tissues behind the peritoneum, finding a potential gap between the peritoneum and the dorsum lumbalis along the fat tissues, separating the gap to reach the abdominal aorta by a cotton swab, separating the abdominal aorta from the inferior vena cava, dissociating an abdominal aorta trunk with the length of about 1cm below the left kidney, measuring the diameter of the renal abdominal aorta by a vernier caliper, and recording;

s2, fully dissociating and ligating branches of the abdominal aorta at the perfusion section by using a silk thread, fully exposing the operation visual field by using a mastoid spreader, blocking the proximal end of the abdominal aorta by using a microscopic vascular clamp, squeezing blood in the blood vessel at the perfusion section to the distal end by using a microscopic hemostatic forceps, blocking the distal end of the abdominal aorta by using the microscopic vascular clamp, fully collapsing the blood vessel to show that the blood vessel has good sealing performance, then extracting 0.2mL of elastase containing 10U by using a disposable insulin syringe, slightly bending the needle head of the syringe by using the vascular forceps to ensure that the syringe slightly takes an L shape, puncturing the abdominal aorta, slowly injecting 0.1mL of elastase containing 5U after successful puncturing, fully filling the abdominal aorta at the perfusion section, fixing the puncture needle, keeping for 20min, timely supplementing if a little elastase oozes outwards during the puncture, keeping the abdominal aorta at the perfusion section in a fully filled state all the time, withdrawing the medicine in the blood vessel after the operation, pulling the needle, covering the puncture point with gelatin sponge, pressurizing with cotton ball, collecting the far-end vascular clamp and the near-end vascular clamp, observing for several minutes until no active hemorrhage occurs, applying the small sliver soaked in 1.0mol/L sterile calcium chloride on the abdominal aorta surface of the perfusion section, taking out the small sliver after 10min, cleaning the peritoneum, cleaning the gap, and closing the incision layer by layer if no hemorrhage occurs.

2. The method for constructing a rat infrarenal abdominal aortic aneurysm model according to claim 1, wherein the measurement of the infrarenal abdominal aorta diameter in step S1 is performed by two persons, and the average value is obtained.

3. The method of claim 2, wherein in step S2, the distance between the vascular clamps is greater than 0.5 cm.

4. The method for constructing a rat infrarenal abdominal aortic aneurysm model by retroperitoneal access according to claim 3, wherein in step S2, the disposable insulin syringe is 1mL 30G.

5. The model of rat infrarenal abdominal aortic aneurysm manufactured by the method for constructing a model of rat infrarenal abdominal aortic aneurysm by retroperitoneal access according to any one of claims 1 to 4.

Technical Field

The invention relates to the technical field of biology and medicine technology, in particular to a rat infrarenal abdominal aortic aneurysm model constructed by retroperitoneal approach and a construction method thereof.

Background

The defects existing in the traditional method for constructing a rat abdominal aortic aneurysm model by combining intracavity elastase perfusion and extracavity calcium chloride infiltration are as follows:

1. the modeling mode is performed through an abdominal cavity approach, the operation is complex, the operation difficulty is high, the risk of abdominal cavity infection exists, the intestinal tract needs to be pulled in the operation, intestinal obstruction is easy to occur, the internal organs in the cavity are damaged, the peritoneum and the posterior peritoneal membrane need to be separated, the operation time is prolonged, and the death risk is increased;

2. the elastase perfusion needs to maintain the perfusion pressure of 100mmHg or more by using a hydraulic or micro-pump injector, but the equal pressure cannot be accurately achieved and maintained in practice, and the overhigh pressure can cause the elastase to enter the circulation to cause the death of rats;

3. the perfusion time is long, 30 minutes to 2 hours are needed, the excessively long abdominal aorta blocking time is long, thrombus is easy to form, and the blood is concentrated above the blocking part for a long time, so that cerebral edema and lower limb ischemia are easy to cause, and the death rate is high;

4. after the perfusion is finished, the iliac artery or the femoral artery on the puncture side needs to be ligated, so that the ischemic necrosis of the unilateral limb is easy to occur, and the disability rate is high.

Disclosure of Invention

In order to solve the technical problems, the invention provides the rat infrarenal abdominal aortic aneurysm model constructed by retroperitoneal approach and the construction method thereof, the operation is relatively simple, the risk caused by the retroperitoneal approach can be avoided, pressurized perfusion is not adopted, the perfusion time is shortened to 20min, no lower limb ischemia occurs, the tumorigenicity rate is not influenced, the damage to blood vessels is extremely small, hemostasis can be realized by local compression after the perfusion is finished, and the success rate of the operation is obviously improved.

The invention relates to a method for constructing a rat infrarenal abdominal aortic aneurysm model by retroperitoneal approach, which comprises the following steps:

s1, fasting the rat before operation for 12 hours, performing pentobarbital sodium anesthesia according to 40-50 mg/kg, taking a right lateral lying position after satisfactory anesthesia, conventionally preparing skin, sterilizing, taking a connecting line from the left axillary posterior line, the costal margin to the thigh root as an incision, wherein the length is 3-3.5 cm, incising skin, subcutaneous tissues and muscles layer by layer to see fat tissues behind the peritoneum, finding a potential gap between the peritoneum and the dorsum lumbalis along the fat tissues, separating the gap to reach the abdominal aorta by a cotton swab, separating the abdominal aorta from the inferior vena cava, dissociating an abdominal aorta trunk with the length of about 1cm below the left kidney, measuring the diameter of the renal abdominal aorta by a vernier caliper, and recording;

s2, fully dissociating and ligating branches of the abdominal aorta at the perfusion section by using a silk thread, fully exposing the operation visual field by using a mastoid spreader, blocking the proximal end of the abdominal aorta by using a microscopic vascular clamp, squeezing blood in the blood vessel at the perfusion section to the distal end by using a microscopic hemostatic forceps, blocking the distal end of the abdominal aorta by using the microscopic vascular clamp, fully collapsing the blood vessel to show that the blood vessel has good sealing performance, then extracting 0.2mL of elastase containing 10U by using a disposable insulin syringe, slightly bending the needle head of the syringe by using the vascular forceps to ensure that the syringe slightly takes an L shape, puncturing the abdominal aorta, slowly injecting 0.1mL of elastase containing 5U after successful puncturing, fully filling the abdominal aorta at the perfusion section, fixing the puncture needle, keeping for 20min, timely supplementing if a little elastase oozes outwards during the puncture, keeping the abdominal aorta at the perfusion section in a fully filled state all the time, withdrawing the medicine in the blood vessel after the operation, pulling the needle, covering the puncture point with gelatin sponge, pressurizing with cotton ball, collecting the far-end vascular clamp and the near-end vascular clamp, observing for several minutes until no active hemorrhage occurs, applying the small sliver soaked in 1.0mol/L sterile calcium chloride on the abdominal aorta surface of the perfusion section, taking out the small sliver after 10min, cleaning the peritoneum, cleaning the gap, and closing the incision layer by layer if no hemorrhage occurs.

Further, in step S1, the diameters of the infrarenal abdominal aorta are measured by two persons, and the average value is obtained.

Further, in step S2, the distance between the two vascular clamps is greater than 0.5 cm.

Further, in the step S2, the specification of the disposable insulin pump is 1mL 30G.

The rat infrarenal abdominal aortic aneurysm model manufactured by the method for constructing the rat infrarenal abdominal aortic aneurysm model is disclosed.

Compared with the prior art, the invention has the beneficial effects that:

1. the retroperitoneal approach is adopted for operation, the retroperitoneal approach is a common operation mode of urinary surgery, and the abdominal aorta is adjacent to the anatomical position of the kidney and belongs to an extraperitoneal organ, so that the possibility is provided for constructing a rat infrarenal abdominal aortic aneurysm model by the retroperitoneal approach, the retroperitoneal approach has the advantages of direct access to the abdominal aorta, no intra-cavity organ interference, relatively simple operation and capability of avoiding risks caused by the retroperitoneal approach;

2. the present invention does not employ pressurized perfusion, since the human abdominal aortic aneurysm formation process also has no external mechanical pressure effect. Only by slowly injecting the elastase into the abdominal aorta at the perfusion section to keep the abdominal aorta full, the model is more suitable for the forming process of the abdominal aortic aneurysm of human beings;

3. the invention shortens the perfusion time to 20min, has no occurrence of 1 case of lower limb ischemia, and does not influence the tumor formation rate;

4. the invention selects the abdominal aorta puncture to recover the original anatomical structure of the abdominal aorta, the selected puncture needle is an insulin needle (30G), the damage to the blood vessel is very small, and the hemostasis can be realized by local compression after the perfusion is finished.

Drawings

FIG. 1 is a flow chart of a method for constructing a rat infrarenal abdominal aortic aneurysm model according to the present invention;

wherein:

a: a surgical incision figure;

b, retroperitoneal space marks retroperitoneal fat maps;

c, a retroperitoneal space entrance diagram;

d, ligating an abdominal aorta collateral branch image of the perfusion section;

e, blocking perfusion segment abdominal aorta diagram

The elastase perfusion map;

g, calcium chloride infiltration graph.

Detailed Description

The following detailed description of embodiments of the present invention is provided in connection with the accompanying drawings and examples. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.

Example (b):

the method comprises the following steps of constructing a rat infrarenal abdominal aortic aneurysm model through retroperitoneal approach:

s1, fasting the rat before operation for 12 hours, performing pentobarbital sodium anesthesia according to 40-50 mg/kg, taking a right lateral lying position after satisfactory anesthesia, conventionally preparing skin, sterilizing, taking a connecting line from the left axillary posterior line, the costal margin to the thigh root as an incision, cutting skin, subcutaneous tissues and muscles layer by layer as shown in A in figure 1, viewing retroperitoneal fat tissues as shown in B in figure 1, finding a potential gap between the peritoneum and the dorsal psoas muscle along the fat tissues as shown in C in figure 1, separating the gap by a cotton swab to reach the abdominal aorta, separating the abdominal aorta from the inferior vena cava, dissociating the abdominal aorta with the length of about 1cm below the left kidney, measuring the diameter of the infrarenal abdominal aorta by a vernier caliper, and recording;

s2, fully dissociating and ligating a branch of the abdominal aorta at the perfusion section by using a mousse line, fully exposing the operation field by using a mastoid spreader as shown in D in figure 1, blocking the proximal end of the abdominal aorta by using a micro vascular clamp, then squeezing blood in the blood vessel at the perfusion section to the distal end by using a micro hemostatic clamp, blocking the distal end of the abdominal aorta by using the micro vascular clamp, completely collapsing the blood vessel, showing that the blood vessel has good tightness as shown in E in figure 1, then extracting 0.2mL containing 10U of elastase by using a disposable insulin syringe, slightly bending the needle head of the syringe by using the vascular clamp, puncturing the abdominal aorta after the blood vessel is slightly in an L shape, slowly injecting 0.1mL containing 5U of elastase after the puncture is successful, fully filling the abdominal aorta at the perfusion section as shown in F in figure 1, fixing the puncture needle, keeping the puncture needle for 20min, and timely supplementing the abdominal aorta at the perfusion section by a little exosmosis of the elastase, withdrawing the intravascular medicine after the operation, pulling out the needle, covering the puncture point with gelatin sponge, pressurizing with a cotton ball, taking the far-end vascular clamp and the near-end vascular clamp, observing for several minutes until no active hemorrhage appears, applying a small yarn strip soaked in 1.0mol/L sterile calcium chloride on the surface of the abdominal aorta of the perfusion section, taking out the small yarn strip after 10 minutes as shown in G in figure 1, cleaning the gap behind the peritoneum, checking if no hemorrhage exists, and closing the incision layer by layer.

As a preferable example of the above embodiment, in step S1, the infrarenal abdominal aorta diameter is measured by two persons and averaged.

As a preferable solution of the above embodiment, in step S2, the distance between the two vascular clamps is greater than 0.5 cm.

As a preferable embodiment of the above embodiment, in step S2, the specification of the disposable insulin syringe is 1mL and 30G.

Comparative example:

the elastase perfusion combined with the calcium chloride infiltration method is a common method for constructing a rat abdominal aortic aneurysm model at present, promotes the formation of the abdominal aortic aneurysm by destroying vascular extracellular matrix and triggering related immune inflammatory reaction to influence the mechanical tension of a vascular wall, and can better simulate the pathological characteristics of the human abdominal aortic aneurysm;

the specific implementation steps are as follows: performing intraperitoneal injection anesthesia with 0.2ml/100g of 3% sodium pentobarbital, positioning in a supine manner, sterilizing, taking a median incision of an abdomen, cutting skin and muscle layer by layer to obtain a length of about 3cm, entering the abdominal cavity, pushing an intestinal canal to the abdominal cavity on the right side, cutting a posterior peritoneum in front of the abdominal aorta, pushing away fat tissues on the front wall and the side wall of the abdominal aorta by using a saline cotton ball, exposing the abdominal aorta, separating a gap between the abdominal aorta and a lower vena cava by using microdissection forceps, dissociating 1cm below the renal artery level and ligating 1-2 pairs of lumbar arteries and inferior mesenteric arteries emitted from a dissociating section by using No. 3-0 silk threads, and blocking the abdominal aorta by clamping the arteries below the renal artery level; about 1c m of free left common iliac artery, ligating the distal end with 3-0 # silk thread, inserting the proximal end into the trocar to a position 0.3cm below the abdominal aorta blocking position, movably pricking the abdominal aorta with 0 # silk thread at the proximal abdominal aorta bifurcation, fixing the trocar, and pressurizing and perfusing the pig pancreas bomb with micro-injection pumpLiprolif (time 30min, speed 4m l/h); simultaneously impregnated with CaCl₂Wrapping the surface of the abdominal aorta with gauze for 10 min; after the perfusion is finished, cutting off a ligature at the far end of the abdominal aorta, removing the trocar, ligating the left common iliac artery, taking down an artery clamp at the near end of the abdominal aorta, and recovering the blood flow of the abdominal aorta; the No. 3-0 silk thread is sutured into the peritoneum, and the No. 0 silk thread closes the abdomen layer by layer.

The above is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several improvements and modifications can be made without departing from the technical principle of the present invention, and these improvements and modifications should also be regarded as the protection scope of the present invention.

The above comparative references are:

[1] establishment and improvement of the model of abdominal aortic aneurysm of Wistar rat [ J ] Med. Proc. of medical research, 2009,22(04):347-349+449.

[2] (Wuyali, child snow shadow, Zhenghong Bourdon, periwinkle. establishment of improved rat abdominal aortic aneurysm model [ J ]. J. China J. Onestology 2014,34(15): 4256-.

[3]Albert Busch,Chernogubova Ekaterina,Jin Hong,et al. Four Surgical Modifications to the Classic Elastase Perfusion Aneurysm Model Enable Haemodynamic Alterations and Extended Elastase Perfusion[J].European Journal of Vascular and Endovascular Surgery,2018,56(1):102-109。