A rabbit internal iliac artery bleeding model and its construction method and application

Abstract

The present invention discloses a method for constructing a rabbit internal iliac artery bleeding model. Under angiographic fluoroscopy, a microguidewire is superselected to the left internal iliac artery of the rabbit, and then a microcatheter is placed in the left internal iliac artery, the microcatheter is fixed, and the microguidewire is withdrawn. The hard end of a V18 guidewire enters the microcatheter to quickly puncture the left internal iliac artery to cause bleeding, and finally angiographic review and whole blood analysis are performed to determine the success of the modeling. This model construction method not only has low trauma and low modeling technical difficulty, but also basically will not cause death due to complications such as infection after surgery, thereby improving the success rate of modeling. In addition, the rabbit internal iliac artery bleeding model constructed by this method can be used in the effect evaluation of interventional vascular embolic materials, which is more objective and accurate, and provides a research tool for the development of new interventional vascular embolic materials and the exploration of their mechanism of action.

Description

Technical Field

[0001] The invention relates to a method for constructing an animal bleeding model, and in particular to a method for constructing a rabbit internal iliac artery bleeding model and application thereof. Background Art

[0002] Currently, the treatments for pelvic arterial bleeding mainly include surgical treatment, interventional embolization and drug therapy. In the past few decades, people have gradually shifted from complex open surgery to interventional treatments with fewer complications and better results, especially vascular interventional embolization. Embolization refers to the injection of embolic agents into target blood vessels for therapeutic purposes to reduce or block blood flow in the target blood vessels to achieve hemostasis or tumor treatment effects.

[0003] In recent years, as embolic materials indispensable for vascular interventional embolization therapy, such as gelatin sponge, polyvinyl alcohol (PVA) particles, sodium alginate and other materials have developed rapidly, but there is still a lack of an ideal animal bleeding model to evaluate the hemostatic performance and application prospects of interventional vascular embolization materials. The animal vascular bleeding model currently used in clinical practice generally uses mice as animal models, finds the target blood vessel through tissue incision, destroys the target blood vessel and then constructs a vascular bleeding model. For example, a Chinese patent (patent number: CN201910157688.7) discloses a method for constructing a rat aortic injury and bleeding animal model and its application in the hemostatic material evaluation system. After the rat is fixed, the unilateral groin is exposed and the skin is prepared; an incision is made at the vertical groin of the external iliac artery on one side, and the subcutaneous fascia and intermuscular connective tissue are bluntly separated layer by layer or not, and the aorta is fully exposed and separated for modeling. However, this open operation is traumatic and the animal is prone to infection complications and death after surgery, which leads to modeling failure; on the other hand, the blood vessels of mice are slender, and it is difficult to perform intravascular intervention. Summary of the invention

[0004] In view of this, the present invention provides a method for constructing a rabbit internal iliac artery bleeding model, which solves the problems of high technical difficulty and low success rate in the existing rat bleeding model construction method.

[0005] The present invention adopts a method for constructing a rabbit internal iliac artery bleeding model, comprising the following steps:

[0006] S101 Animal Anesthesia:

[0007] The experimental rabbits were fasted before operation, anesthesia was induced by intramuscular injection of Sumenxin, and anesthesia was maintained by intravenous injection of Zotal.

[0008] S102 leaks the left common carotid artery:

[0009] A longitudinal incision was made at the strongest pulsation point of the left common carotid artery, and the left common carotid artery was bluntly separated using an elbow hemostatic forceps, and suspended with 2-0 mousse sutures at the proximal and distal ends;

[0010] S103 Construction of left internal iliac artery bleeding model:

[0011] First, the left common carotid artery was punctured with a 21G micro-needle puncture needle, and then a V18 guidewire was inserted to exchange the microcatheter. 5 mL of iohexol was quickly injected above the bifurcation of the abdominal aorta using a 5 mL syringe to observe the course and branches of the bilateral iliac arteries of the New Zealand female rabbits. The micro-guidewire was then superselected to the left internal iliac artery, and then the microcatheter was placed in the left internal iliac artery, the microcatheter was fixed, the micro-guidewire was withdrawn, and the hard end of the V18 guidewire was inserted into the microcatheter to quickly puncture the left internal iliac artery to cause bleeding. Finally, angiography was repeated and whole blood analysis was performed to evaluate whether the model was successful.

[0012] The method for constructing a rabbit internal iliac artery bleeding model provided by the present invention, on the one hand, selects the rabbit internal iliac artery as the puncture route, which solves the problem that the diameter of the rabbit femoral artery is less than 1mm and the puncture and catheterization are difficult; on the other hand, under the fluoroscopic angiography, the microguidewire is superselected to the left internal iliac artery, and then the microcatheter is placed in the left internal iliac artery, the microcatheter is fixed, the microguidewire is withdrawn, and the hard end of the V18 guidewire enters the microcatheter to quickly puncture the left internal iliac artery to cause bleeding. This modeling method not only has little trauma, low modeling technical difficulty, but also basically will not cause death due to complications such as infection after surgery, improves the success rate of modeling, and reduces the error of individual differences in modeling, and has high stability.

[0013] Furthermore, the rabbit internal iliac artery bleeding model constructed by this method can be used to evaluate the effects of interventional vascular embolic materials, which is more objective and accurate, and provides a research tool for the development of new interventional vascular embolic materials and the exploration of their mechanisms of action. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Schematic diagram of the exposure of the left common carotid artery of a New Zealand rabbit after percutaneous incision;

[0015] Figure 2 This is angiography of the lower abdominal aorta of a New Zealand rabbit, showing the iliac artery and arteries of both lower limbs completely;

[0016] Figure 3 This is an angiographic image of the rabbit internal iliac artery bleeding model constructed in Example 1 of the present invention, wherein Figure 3 A: Normal angiography. Before modeling, the left internal iliac artery had a normal course and no contrast agent leakage. Figure 3 B: Left internal iliac artery bleeding model. After modeling, the left internal iliac artery has overflowed with contrast agent and bleeding can be seen.

[0017] Figure 4The changes of red blood cell count, hematocrit and hemoglobin content before and after the rabbit internal iliac artery bleeding model established by the method of Example 1 of the present invention. Note: n = 8, x ± S, * indicates that compared with before modeling, *P < 0.05, **P < 0.01;

[0018] Figure 5 This is an angiographic image to evaluate the effect of using gelatin sponge as an interventional vascular embolization material in a rabbit internal iliac artery hemorrhage model. It can be seen that the bilateral internal iliac arteries were truncated and the signs of contrast agent overflow disappeared. DETAILED DESCRIPTION

[0019] The principles and features of the present invention are described below in conjunction with the accompanying drawings. The examples given are only used to explain the present invention and are not used to limit the scope of the present invention.

[0020] The experimental animals involved in the present invention are: New Zealand female rabbits, 3 months old, weighing 8-10 kg, Changsha Tianqin, license number: SCXK (Xiang) 2019-0015.

[0021] The experimental instruments and materials involved in the present invention include: digital subtraction angiography machine (General Electric, USA), 21G puncture needle (COOK, USA), V18 guide wire (Boston Scientific), gelatin sponge block (Ailikang), microcatheter and microguide wire (Boston Scientific), iohexol injection (General Electric, USA), Suminxin II (Dunhua Shengda), Shutai 50 (Vick, France), and fully automatic blood cell analyzer (Mindray 6800plus).

[0022] Embodiment 1: A method for constructing a rabbit internal iliac artery bleeding model, specifically comprising the following steps:

[0023] S101 Animal Anesthesia:

[0024] Female New Zealand rabbits were fasted for 12 h before surgery. After anesthesia was induced by intramuscular injection of Suminxin II (2 mg / kg), anesthesia was maintained by intraauricular injection of Zoltrate (7.5 mg / kg).

[0025] S102 leaks the left common carotid artery:

[0026] After the experimental rabbits were anesthetized successfully, the neck skin was prepared and disinfected, and a longitudinal incision of about 2 cm was made at the skin where the left common carotid artery pulsated the strongest. The left common carotid artery was bluntly separated using an elbow hemostatic forceps (12.5 cm) and suspended with 2-0 mousse thread at the proximal and distal ends.

[0027] S103 Construction of left internal iliac artery bleeding model:

[0028] After successful puncture of the left common carotid artery with a 21G puncture needle, a V18 guidewire exchange microcatheter was inserted. 5 mL of iohexol was quickly injected above the bifurcation of the abdominal aorta using a 5 mL syringe. The course and branching of the bilateral iliac arteries of the New Zealand female rabbits were observed. The microguidewire was then superselected to the left internal iliac artery, and the microcatheter was placed in the left internal iliac artery. The microcatheter was fixed and the microguidewire was withdrawn. The hard end of the V18 guidewire was inserted into the microcatheter to quickly puncture the left internal iliac artery. Angiography was performed to review and evaluate whether the modeling was successful.

[0029] The model established by the method of Example 1 was evaluated for its effect, and the results are as follows:

[0030] (1) Observation and statistics of modeling results: The model was successfully established in 8 New Zealand rabbits using the method of Example 1, with a success rate of 100%. There was no infection, no death, no hematuria or bloody stool, and no complications of cerebral infarction such as hemiplegia and lip deviation. 1-3 days after modeling, the New Zealand rabbits had a decreased appetite, poor mental state, and were lazy to move. However, after 3 days, their appetite and mental state gradually returned to normal and they could walk freely.

[0031] (2) Figure 2 The results showed that by using a microcatheter placed in the lower segment of the abdominal aorta for angiography, the bilateral common iliac arteries were visualized. The internal iliac artery was the first branch of the common iliac artery, which continued downward to the external iliac artery, common femoral artery, deep femoral artery, superficial femoral artery, popliteal artery, anterior tibial artery, posterior tibial artery and peroneal artery. The microguidewire was shaped to allow the microcatheter to be super-selected into the bilateral internal iliac arteries for modeling.

[0032] (3) Figure 3 The angiography results showed that the bilateral internal iliac arteries had normal course without abnormal stenosis or tumor-like dilatation before the hemorrhage model was made. Figure 3 A); The microcatheter was placed in the left internal iliac artery, and the left internal iliac artery was punctured with the hard end of the V18 guidewire. The angiography showed bleeding signs of contrast agent overflow in the left internal iliac artery ( Figure 3 B), indicating that the rabbit internal iliac artery bleeding model was successfully established.

[0033] (4) Figure 4 The results showed that 5 mL of whole blood was drawn from the ear vein before and 10 minutes after modeling and placed in a blood collection tube containing EDTA-K2 anticoagulant, and the red blood cell count, hematocrit, and hemoglobin content were measured by a blood cell analyzer. The results of venous whole blood analysis of rabbits before and after modeling further showed that the red blood cell count, hemoglobin, and hematocrit of rabbits were significantly reduced after modeling, indicating that the bleeding model was successfully established.

[0034] Example 2: Evaluation of the effect of gelatin sponge interventional embolization using the rabbit internal iliac artery bleeding model of Example 1

[0035] After the modeling is successfully established in Example 1, wait for 10 minutes, superselect the bilateral internal iliac arteries with microcatheter microguidewires, cut the gelatin sponge into 3x10 mm in size, roll the gelatin sponge into 0.5 mmx10 mm in size on a dry gauze piece, place it in a 1 mL syringe, and quickly inject it into the bilateral internal iliac artery trunks through the microcatheter. After angiography, the bilateral internal iliac arteries were cut off, and the signs of contrast agent overflow disappeared. The microcatheter was removed, and the gelatin sponge was pressed with appropriate force on the left common carotid artery for 15 minutes. Figure 5 The results showed that the bilateral internal iliac arteries were cut off and the bleeding signs disappeared. The experimental results showed that the hemostasis was successful through the embolization of gelatin sponge blocks. There was no infection, death, cerebral infarction and other complications in the New Zealand rabbits after the embolization. It can be determined that the gelatin sponge material can be further studied and applied as an excellent vascular embolization material.

[0036] In summary, the method for constructing the rabbit internal iliac artery bleeding model provided by the present invention is adopted. Under the fluoroscopic angiography, the micro-guidewire is superselected to the left internal iliac artery, and then the micro-catheter is placed in the left internal iliac artery, the micro-catheter is fixed, the micro-guidewire is withdrawn, and the hard end of the V18 guidewire enters the micro-catheter to quickly puncture the left internal iliac artery to cause bleeding. This modeling method is not only less traumatic and has low technical difficulty, but also basically will not cause death due to complications such as infection after surgery, improves the success rate of modeling, and reduces the error of individual differences in modeling, and has high stability.

[0037] Furthermore, the rabbit internal iliac artery bleeding model of the present invention is applied to the evaluation of the effect of interventional vascular embolic materials, which is more objective and accurate, and provides a research tool for the development of new interventional vascular embolic materials and the exploration of their mechanism of action.

[0038] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A rabbit internal iliac artery bleeding model, characterized in that: The model is constructed by the following method: S101 Animal Anesthesia: The experimental rabbits were fasted before operation, anesthesia was induced by intramuscular injection of Sumenxin, and anesthesia was maintained by intravenous injection of Zotal. S102 Exposure of the left common carotid artery: A longitudinal incision was made at the strongest pulsation point of the left common carotid artery, and the left common carotid artery was bluntly separated using an elbow hemostatic forceps, and suspended with 2-0 mousse sutures at the proximal and distal ends; S103 Construction of left internal iliac artery bleeding model: First, the left common carotid artery was punctured with a puncture needle, and then a V18 guidewire exchange microcatheter was inserted. 5 mL of iohexol was quickly injected above the bifurcation of the abdominal aorta using a 5 mL syringe to observe the course and branches of the bilateral iliac arteries of the New Zealand female rabbits. The microguidewire was then superselected to the left internal iliac artery, and then the microcatheter was placed in the left internal iliac artery. The microcatheter was fixed and the microguidewire was withdrawn. The hard end of the V18 guidewire was inserted into the microcatheter to quickly puncture the left internal iliac artery to cause bleeding. Finally, angiography was repeated and whole blood analysis was performed to evaluate whether the model was successful.

2. Use of the rabbit internal iliac artery bleeding model described in claim 1 in evaluating the effects of interventional vascular embolization materials.

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