Method for constructing rat model through femoral artery hepatic arteriography and perfusion chemotherapy embolism

Abstract

The invention relates to a method for constructing a rat model for hepatic artery angiography and perfusion chemotherapy embolization through femoral artery, and relates to a novel hepatic artery angiography and perfusion chemotherapy embolization method which is established through research, in particular to a hepatic artery intubation angiography and chemotherapy embolization method for performing super selection through a rat femoral artery approach. The method is different from the traditional intubation angiography and chemoembolization which are retrograde from laparotomy to hepatic artery through stomach duodenal artery. According to the method, exposed wounds are obviously reduced, the infection incidence rate of the rats is reduced, the bleeding amount of the rats is reduced, and the success rate of an operation is increased. The injection of chemotherapy drugs and embolism particles on the liver tumor of the rat is carried out through the femoral artery path of the rat, so that the scientific research on the liver tumor of the rat through different treatment means is conveniently carried out and verified, the success rate of an operation is increased, related complications of the rat are reduced, and the requirements of large-scale related experimental research can be met.

Description

Technical Field

[0001] The present disclosure relates to the field of clinical medical technology, and in particular to a method for constructing a rat model of hepatic artery angiography and perfusion chemotherapy embolization via the femoral artery and its application. Background Art

[0002] At present, the main treatment methods for primary hepatocellular carcinoma are: surgical resection, liver transplantation, transarterial chemoembolization (TACE), stereotactic radiotherapy, chemotherapy, local ablation, biological therapy, etc. Surgical resection is the first choice for liver cancer, especially for small liver cancer, the 5-year survival rate after surgery is as high as about 90%, but most patients are diagnosed with liver cancer in the middle and late stages and cannot be surgically removed.

[0003] Hepatic artery angiography chemoembolization (TACE) has little effect on normal tissue blood circulation and is currently the preferred treatment for unresectable liver cancer. It is also widely used as an effective method to prevent and treat tumor recurrence and metastasis after tumor resection. Some patients with large tumors that cannot be resected in the first stage can also undergo this method to downstage the tumor and then undergo a second-stage surgical resection. At the same time, TACE treatment can also reduce the risk of spontaneous rupture and bleeding of the tumor. However, the overall effectiveness of TACE treatment is about 70%-80%, and there is still uncertainty in the treatment effect, and there is still room for further improvement in efficacy.

[0004] In view of the wide application and limitations of TACE treatment, it is urgent to correct its methods and explore new methods or embolic materials to improve the effectiveness of TACE treatment. The research of new methods requires the use of a large number of basic animals to verify the effectiveness and advantages of treatment, and at the same time, it is necessary to continuously improve and develop the technology and methods of TACE.

[0005] The catheterization angiography and perfusion chemoembolization through the rat hepatic artery is the basic method and the best choice of TACE. At present, the main method for catheterization angiography and embolization chemotherapy of the rat hepatic artery is to open the abdomen under anesthesia, expose and free the proper hepatic artery and gastroduodenal artery, temporarily clamp the hepatic artery, cut the gastroduodenal artery under direct vision, and retrogradely catheterize the proper hepatic artery and ligate and fix the proper hepatic artery. Although this experimental method has a certain degree of adoption, its steps are cumbersome and complicated, and it is difficult to meet the requirements of large-scale related experimental research. Summary of the invention

[0006] In order to solve the above problems, the present application proposes a method for constructing a rat model of hepatic artery angiography and perfusion chemotherapy embolization via the femoral artery and its application.

[0007] In one aspect, the present application proposes a method for constructing a rat model of hepatic artery angiography and perfusion chemotherapy embolization via the femoral artery, comprising the following steps:

[0008] Prepare rat models and precondition;

[0009] The femoral artery and vein area of ​​rats were skin prepared;

[0010] Cut the subcutaneous tissue at the skin preparation site, separate and expose the femoral artery;

[0011] Use the needle to insert the microguidewire into the femoral artery and pre-tighten the proximal end of the femoral artery;

[0012] Along the exposed micro-guidewire, the pre-constructed micro-catheter is sent into the femoral artery and the proximal end of the femoral artery is loosened; under the fluoroscopy of the DSA machine, the micro-catheter is passed over the common iliac artery to the lower segment of the abdominal aorta, and the micro-catheter is fixed; the guidewire retained in the middle and upper segments of the abdominal aorta is super-selectively sent to the celiac trunk, and the micro-guidewire is fixed; the micro-catheter retained in the lower segment of the abdominal aorta is then retrogradely super-selectively sent to the proper hepatic artery along the guidewire direction, and the micro-catheter is fixed; the micro-guidewire is withdrawn; iohexol contrast agent is injected from the end of the micro-catheter using a 1 ml insulin needle, angiography is performed, and chemotherapy drugs and embolic materials are selectively injected, so as to obtain the rat model of hepatic artery (catheterization) angiography and perfusion chemotherapy embolization via the femoral artery;

[0013] Repeat the above steps as needed;

[0014] When the preset conditions are met, the microcatheter is removed.

[0015] On the other hand, the present application proposes a method for constructing a rat model of hepatic arteriography and perfusion chemoembolization via the femoral artery for application in primary hepatocellular carcinoma surgery.

[0016] Technical effects of the present invention:

[0017] This application establishes a new method of transhepatic artery angiography and chemoembolization through research. This study conducts superselective hepatic artery catheterization angiography and chemoembolization via the rat femoral artery, which is different from the traditional transabdominal gastroduodenal artery retrograde hepatic artery catheterization angiography and chemoembolization. This method not only significantly reduces the exposed wounds and reduces the incidence of infection in rats, but also reduces the amount of bleeding in rats and increases the success rate of surgery. Injection of chemotherapy drugs and embolic particles into rat liver tumors via the rat femoral artery is carried out to facilitate and verify scientific research on rat liver tumors through different treatment methods, which not only increases the success rate of surgery and reduces related complications in rats, but also meets the requirements of large-scale related experimental research.

[0018] Further features and aspects of the present disclosure will become apparent from the following detailed description of exemplary embodiments with reference to the attached drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate exemplary embodiments, features, and aspects of the disclosure and, together with the description, serve to explain the principles of the disclosure.

[0020] Figure 1 Shown is a schematic diagram of a longitudinal section of the clinical operation of the present invention;

[0021] Figure 2 Shown is a schematic diagram of the clinical operation of separation and ligation of the femoral artery and vein of rats according to the present invention;

[0022] Figure 3 Shown is a schematic diagram of the clinical operation of delivering a guidewire catheter into a femoral artery according to the present invention;

[0023] Figure 4 Shown is a schematic diagram of the development of hepatic artery angiography of the present invention;

[0024] Figure 5 Shown is a schematic diagram of a rat model after wound suture of the present invention;

[0025] Figure 6 Shown is a schematic diagram of the clinical operation of exposing the rat liver after laparotomy in the traditional method;

[0026] Figure 7 The figure shows a clinical operation diagram of cutting a small incision in the lower part of the gastroduodenal artery and inserting a catheter into the femoral artery through a guidewire catheter in a retrograde manner in the traditional method;

[0027] Figure 8 The figure shows the clinical operation diagram of intra-abdominal infection and pus discharge in rats after open abdominal intubation in the traditional method. DETAILED DESCRIPTION

[0028] Various exemplary embodiments, features and aspects of the present disclosure will be described in detail below with reference to the accompanying drawings. The same reference numerals in the accompanying drawings represent elements with the same or similar functions. Although various aspects of the embodiments are shown in the accompanying drawings, the drawings are not necessarily drawn to scale unless otherwise specified.

[0029] The word “exemplary” is used exclusively herein to mean “serving as an example, example, or illustration.” Any embodiment described herein as “exemplary” is not necessarily to be construed as preferred or advantageous over other embodiments.

[0030] In addition, in order to better illustrate the present disclosure, numerous specific details are given in the following specific embodiments. It should be understood by those skilled in the art that the present disclosure can also be implemented without certain specific details. In some examples, means, components and circuits well known to those skilled in the art are not described in detail in order to highlight the main purpose of the present disclosure.

[0031] The rat model used in this embodiment can be provided by the laboratory.

[0032] The experimental equipment, instruments and reagents used in this embodiment can be provided by the laboratory or purchased from the market.

[0033] The present invention establishes a new cannulation pathway, and after cannulation into the hepatic artery via the femoral artery of rats, simulates the human body's transcatheter arterial chemoembolization (TACE) to perform various drug perfusion chemotherapy, targeted, immune and other drugs on rats, or inject embolic materials or related new synthetic materials (such as nanoparticles, etc.), so as to facilitate scientific research on rat liver tumors through different treatment methods.

[0034] The proposed method for constructing a rat model of hepatic arteriography and perfusion chemoembolization via femoral artery has the following main implementation steps:

[0035] 1. General anesthesia and skin preparation

[0036] Rats were anesthetized with intraperitoneal injection of 0.8 ml of pentobarbital at a concentration of 20 mg / ml (rats weighed 400 g ± 50 g before surgery). After the rats were deeply sedated, the inguinal area was prepared with a razor, with a range of about 3 cm*3 cm, and the inguinal area (femoral artery and vein area) of the rats was exposed;

[0037] 2. Separate and expose the femoral artery

[0038] Make a longitudinal skin incision along the groin, with a length of approximately 3 cm ( Figure 1 ), use forceps to separate subcutaneous tissue layer by layer, expose and slowly separate femoral artery and vein, separate femoral artery and vein with 5cm 3-0 suture, then use the same length of 3-0 suture to pass through the lower edge of femoral artery and ligate the distal end of femoral artery, and at the same time, use the same length of 3-0 suture to pass through the femoral artery and slightly lift the proximal end, so that slight pulsation of femoral artery can be seen, and fully expose about 2cm of femoral artery (see Figure 2 );

[0039] 3. Construct an artificial rat femoral artery sheath and insert the guidewire catheter

[0040] Pre-soak a 0.014-inch micro-guidewire (30 cm long) and micro-catheter in heparinized 0.9% sodium chloride saline. Use the micro-guidewire to penetrate the tail of the needle of a 2 ml syringe, and use the needle to slightly puncture the exposed distal end of the femoral artery. When bright red blood spurts out, immediately tighten the proximal suture. Under the fluoroscopy of the DSA machine, gradually push the guidewire in the needle retrogradely into the abdominal aorta, then slowly withdraw the needle and leave the micro-guidewire in the abdominal aorta (see Figure 3 ), to prevent bleeding from the puncture wound, continue to slightly tighten the proximal suture.

[0041] 4. The microcatheter is super-selectively inserted into the hepatic artery along the guidewire for treatment

[0042] A 1.7F microcatheter (total length 20 cm) that has been flushed with heparinized saline is slowly inserted along the exposed end of the indwelling microguidewire through the femoral artery rupture (the opening punctured by a 2 cm needle). When the microcatheter successfully enters the femoral artery rupture and the rupture stops bleeding, the proximal suture is loosened.

[0043] Under DSA fluoroscopy, the microcatheter is passed over the common iliac artery to the lower abdominal aorta, and then fixed. The guidewire placed in the upper and middle abdominal aorta is super-selectively sent into the hepatic artery by continuously adjusting the direction of the guidewire, and then the microguidewire is fixed.

[0044] The microcatheter placed in the lower segment of the abdominal aorta is retrogradely superselected along the guidewire to the hepatic artery. After the position of the microcatheter is fixed, the microguidewire is slowly withdrawn out of the body and continues to be placed in heparinized saline.

[0045] Use a 1ml insulin syringe to extract iohexol contrast agent, insert the needle into the end of the microcatheter for manual push contrast imaging (the total amount of contrast imaging is controlled at 0.5-1ml each time), and the branches of the hepatic artery (left, middle, right branches and their small branches) can be clearly seen after contrast imaging. After the microcatheter is in place, various chemotherapy drugs can be injected through the 1ml insulin syringe, or embolic materials or related new synthetic materials (such as nanoparticles, etc.) can be injected (see Figure 4 ).

[0046] 5. Remove the microcatheter and suture the wound

[0047] After the injection of drugs or materials is completed, the contrast agent is manually pushed again to observe the blood vessels in the hepatic artery and its branches. After the injection is completed, the microcatheter is slowly removed, and the 3-0 suture that was previously passed through is immediately used to ligate the proximal femoral artery. Observe whether there is bleeding at the puncture point. If there is no bleeding, it indicates that the femoral artery in the surgical area has been ligated successfully. Finally, 3-0 sutures are used with 7x20 triangular suture needles to suture the inguinal wound, and then the abdominal wall is routinely disinfected with alcohol wipes, fixed with medical tape, and the rat is returned to the cage after waking up. (See Figure 5 )

[0048] In order to compare with the traditional method, the present invention constructed an experimental group (Group A: 30 rats) and a control group (Group B: 30 rats). Group A and Group B were treated by the present method and the traditional method of transabdominal transgastric artery. The results are shown in Table 1 below:

[0049]

[0050] Table 1---Comparison data table

[0051] The success rate of superselective hepatic artery catheterization and chemoembolization via the femoral artery of rats was 90.0% (27 / 30). Retrograde catheterization failed in 2 rats due to the variation of the hepatic artery course, and in 1 rat, the microcatheter could not be inserted due to the thin femoral artery. However, it was successfully inserted into the hepatic artery for angiography after another test from the contralateral femoral artery one week later. No wound infection, postoperative hemorrhage, or abnormal movement of the lower limbs of rats occurred after surgery. The average amount of blood loss after surgery in 30 rats was less than 1 ml.

[0052] The success rate of the traditional method of transabdominal catheterization and chemoembolization via retrograde gastroduodenal artery to hepatic artery was 63.3% (19 / 30 of 30 cases were operated on using the traditional method). Figure 6 and Figure 7 As shown in the figure, the main reasons for failure of the traditional method are: unsuccessful retrograde cannulation due to the variation of the hepatic artery; failure to cannulate successfully due to damage to the gastroduodenum and hepatic artery caused by insufficiently careful operation; death due to bleeding at the puncture site after the cannulation was completed; and death due to infection caused by wound tearing and intestinal exposure on the second day after surgery (see Figure 8 The average postoperative bleeding volume of 30 rats was 5 ml. Please understand the traditional method in conjunction with the background technology.

[0053] In contrast, the present invention obviously has the following technical advantages:

[0054] 1. Significantly reduce the size of the exposed wound. The exposed wound of 3 cm is significantly smaller than the diameter of the traditional anatomical wound of more than 5 cm. At the same time, it is an extracorporeal operation (groin area). The traditional method is an open abdominal operation. Figure 1 and Figure 7 contrast)

[0055] 2. Reduce the incidence of postoperative infection in rats: None of the 30 rats developed infection after surgery, while 4 rats died of infection after surgery using the traditional method (4 / 30). This was considered to be caused by intraperitoneal infection after intestinal separation by in vivo manipulation ( Figure 5 and Figure 8 contrast);

[0056] 3. Reduce the amount of bleeding in rats (the average amount of bleeding after surgery for 30 rats was less than 1 ml, while the average amount of bleeding after surgery for 30 rats with the traditional method was 5 ml, and hemostasis was more complicated) Figure 2 and Figure 6 contrast);

[0057] 4. Increase the success rate of surgery (90% (27 / 30) of surgery was successful, 63.3% (19 / 30) of traditional surgery). The success rate of catheterization via the femoral artery was significantly higher than that via the retrograde cannulation via the gastroduodenal artery (χ2=4.565, P=0.033), which was statistically significant (P<0.05).

[0058] Therefore, compared with the traditional open abdominal gastroduodenal artery retrograde to hepatic artery catheterization and chemoembolization, the present invention has the following advantages:

[0059] 1. Significantly reduce the size of the exposed wound. The exposed wound of 3 cm is significantly smaller than the diameter of the traditional anatomical wound of more than 5 cm. At the same time, it is an extracorporeal operation (groin area), while the traditional method is an open abdominal operation.

[0060] 2. Reduce the incidence of postoperative infection in rats (no infection occurred in 30 rats after surgery, while 4 rats died of infection after surgery using the traditional method (4 / 30), which was considered to be an intraperitoneal infection after intestinal separation by in vivo manipulation);

[0061] 3. Reduce the amount of bleeding in rats (the average amount of bleeding after surgery for 30 rats was less than 1 ml, while the average amount of bleeding after surgery for 30 rats using the traditional method was 5 ml, and hemostasis was more complicated);

[0062] 4. Increase the success rate of surgery (90% (27 / 30) of surgery was successful, 63.3% (19 / 30) of traditional surgery). The success rate of cannulation via the femoral artery was significantly higher than that via the retrograde cannulation via the gastroduodenal artery (χ2=4.565, P=0.033), which was statistically significant (P<0.05);

[0063] 5. Study the therapeutic effects of relevant new materials (such as nanoparticles of different diameters) and new drugs (new targeted drugs, immune drugs, etc.) on liver tumors through the femoral artery approach and compare them with relevant clinical studies;

[0064] 6. Bilateral renal artery, superior mesenteric artery, abdominal aorta angiography and a series of related treatments can also be performed through the femoral artery approach;

[0065] 7. Through the femoral artery approach, the intrahepatic vascular conditions after drug injection can be directly observed under DSA fluoroscopy, which is not possible with the traditional approach.

[0066] Therefore, the present invention performs angiography and chemoembolization by puncturing the femoral artery of rats and super-selectively inserting a guidewire catheter to the hepatic artery. This is different from the traditional laparotomy and retrograde catheterization of the hepatic artery through the gastroduodenal artery for angiography and chemoembolization. It not only increases the success rate of the operation and reduces related complications in rats, but also can meet the requirements of large-scale related experimental research.

[0067] The embodiments of the present disclosure have been described above, and the above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and changes will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The selection of terms used herein is intended to best explain the principles of the embodiments, practical applications, or technical improvements in the market, or to enable other persons of ordinary skill in the art to understand the embodiments disclosed herein.

Claims

1. A method for constructing a rat model of hepatic artery angiography and perfusion chemotherapy embolization via the femoral artery, characterized in that: The steps include: Prepare rat models and precondition; The femoral artery and vein area of ​​rats were skin prepared; Cut the subcutaneous tissue at the skin preparation site, separate and expose the femoral artery; Use the needle to insert the microguidewire into the femoral artery and pre-tighten the proximal end of the femoral artery; Along the exposed micro-guidewire, the pre-constructed micro-catheter is sent into the femoral artery and the proximal end of the femoral artery is loosened; under the fluoroscopy of the DSA machine, the micro-catheter is passed over the common iliac artery to the lower segment of the abdominal aorta, and the micro-catheter is fixed; the guidewire retained in the middle and upper segments of the abdominal aorta is super-selectively sent to the celiac trunk, and the micro-guidewire is fixed; the micro-catheter retained in the lower segment of the abdominal aorta is then retrogradely super-selectively sent to the proper hepatic artery along the direction of the guidewire, and the micro-catheter is fixed; the micro-guidewire is withdrawn; iohexol contrast agent is injected from the end of the micro-catheter using a 1 ml insulin needle, angiography is performed, and chemotherapy drugs and embolic materials are selectively injected, so as to obtain the rat model of hepatic artery angiography and perfusion chemotherapy embolization via the femoral artery; Repeat the above steps as needed; When the preset conditions are met, the microcatheter is removed.

2. The method for constructing a rat model of hepatic artery angiography and perfusion chemoembolization via femoral artery according to claim 1, characterized in that: The pretreatment of the rat model includes: using 0.8 ml of pentobarbital with a concentration of 20 mg / ml to perform general anesthesia injection through the abdominal cavity.

3. The method for constructing a rat model of hepatic artery angiography and perfusion chemotherapy embolization via femoral artery according to claim, characterized in that: The micro guide wire and micro catheter are pre-soaked in heparinized 0.9% sodium chloride saline.

4. The method for constructing a rat model of hepatic artery angiography and perfusion chemoembolization via femoral artery according to claim 1, characterized in that: The micro guide wire adopts a 0.014in model.

5. The method for constructing a rat model of hepatic artery angiography and perfusion chemoembolization via femoral artery according to claim 1, characterized in that: The microcatheter adopts 1.7F model.

6. Research on the application of the method for constructing a rat model of hepatic artery angiography and perfusion chemoembolization via femoral artery according to any one of claims 1 to 5 in primary hepatocellular carcinoma surgery.

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