Mouse reversible ureteral obstruction model and construction method thereof

Abstract

The invention discloses a mouse reversible ureteral obstruction model and a construction method thereof, and the construction method comprises the following steps: carrying out anesthesia treatment on a mouse, fixing the supine position of the mouse, and carrying out skin preparation disinfection; cutting the middle of the lower abdomen of the mouse, removing the bladder out of the body, and exposing a ureter area and the joint of the lower end of the free ureter and the ureter bladder; the bladder tissue below the ureter bladder connecting position is ligated; sewing and closing abdominal muscles and skin by adopting a suture line; according to the construction method disclosed by the invention, through setting of a specific construction method, an obstruction region can be formed in a mouse bladder region, and the ureter cannot be obviously damaged, so that the obstruction success rate of the model can reach 100%, and the success rate of a recanalization operation can reach 90% or above.

Description

Technical Field

[0001] This invention relates to the field of ureteral obstruction model construction technology, specifically to a mouse reversible ureteral obstruction model and its construction method. Background Technology

[0002] Renal interstitial fibrosis is a major pathological change in progressive chronic kidney disease and a leading factor in the gradual loss of kidney function. A better understanding of its pathogenesis and the development of effective treatments may reduce the global burden of end-stage renal disease. Unilateral ureteral obstruction (UUO) is a simple, stable, and reliable model widely used in renal fibrosis research. Reversible UUO (R-UUO) has been used to study the structural and functional recovery of the kidney after obstruction relief and shows great potential in the study of inflammatory and immune processes, cell and tissue regeneration, and subsequent tissue remodeling. Mouse R-UUO models play an important role due to their well-defined genetic background, close resemblance to humans, and ease of experimental cost.

[0003] Currently, there are various methods for creating R-UUO models, such as non-invasive microvascular clamping, polyethylene (silicone) tube wrapping, polyethylene tube clamping, and ureterovesical anastomosis. Because the mouse ureter is very thin, with an inner diameter of less than 0.1 mm, both external ureteral compression to create obstruction and reimplantation can cause ureteral damage, leading to recanalization failure or incomplete recanalization.

[0004] In view of this, the present invention is hereby proposed. Summary of the Invention

[0005] The purpose of this invention is to provide a mouse reversible ureteral obstruction model and its construction method. The construction method of this invention forms an obstruction area in the bladder region, and through the setting of specific construction methods, the success rate of ureteral obstruction model construction can reach 100%, and the success rate of recanalization surgery can reach more than 90%.

[0006] In order to achieve the above-mentioned objectives of the present invention, the following technical solution is adopted:

[0007] The first aspect of this invention provides a method for constructing a mouse model of reversible ureteral obstruction, the method comprising the following steps:

[0008] (a) Anesthetize the mice, then fix them in a supine position and prepare and disinfect their skin;

[0009] (b) Make an incision in the middle of the lower abdomen of the mouse, remove the bladder from the body and expose the ureteral area and the freed lower end of the ureter and the ureterovesical junction;

[0010] (c) Ligate the bladder tissue below the ureterovesical junction;

[0011] (d) The abdominal muscles and skin were closed by suturing with sutures.

[0012] Preferably, the ligation of the bladder tissue below the ureterovesical junction includes clamping with a vascular clamp; ligation with a suture; or removing a section of silicone tube and then passing a suture through the silicone tube to ligate the bladder tissue.

[0013] Preferably, in step (a), the anesthesia is performed by intraperitoneal injection of 1.0% sodium pentobarbital at a dose of 50 mg / kg into mice.

[0014] Preferably, in step (b), the incision length is 0.6 to 1.5 cm.

[0015] Preferably, in step (b), the length of the ureter freed is 0.3 to 0.5 cm.

[0016] Preferably, in step (c), the silicone tube has a length of 2.5 to 3.5 mm, an inner diameter of 1.2 to 1.8 mm, and an outer diameter of 2.2 to 2.8 mm.

[0017] Preferably, in step (d), the sutures are all 5-0 sutures.

[0018] Preferably, the knot is tied during the ligation process to avoid the ureterovesical junction and the urethra.

[0019] Preferably, the construction method has a 100% success rate.

[0020] A second aspect of the present invention provides a mouse model of reversible ureteral obstruction in which the obstruction region is located in the bladder, constructed by the above-described method.

[0021] Compared with the prior art, the beneficial effects of the present invention include at least the following:

[0022] The construction method of this invention, through specific construction method settings, can form an obstruction area in the bladder region of mice without causing significant damage to the ureter, so that the success rate of model obstruction construction can reach 100% and the success rate of recanalization surgery can reach more than 90%. It avoids the problems of ureteral damage that may be caused by the external compression of the ureter in existing technologies and the reimplantation method. It effectively solves the problem that the existing technologies are prone to recanalization failure or incomplete recanalization. Attached Figure Description

[0023] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the accompanying drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, the elements or parts are not necessarily drawn to scale.

[0024] Figure 1 These are practical diagrams illustrating the construction process of the mouse reversible ureteral obstruction model in this embodiment of the invention.

[0025] Figure 2 These are ultrasound images of mice before, after, and after recanalization in the experimental examples of this invention. Detailed Implementation

[0026] The embodiments of the technical solution of the present invention will be described in detail below with reference to the examples. The following embodiments are only used to illustrate the technical solution of the present invention more clearly, and are therefore only examples, and should not be used to limit the scope of protection of the present invention.

[0027] It should be noted that, unless otherwise stated, the technical or scientific terms used in this application should have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.

[0028] This invention provides a method for constructing a mouse model of reversible ureteral obstruction, the method comprising the following steps:

[0029] (a) Anesthetize the mice, then fix them in a supine position and prepare and disinfect their skin;

[0030] (b) Make an incision in the middle of the lower abdomen of the mouse, remove the bladder from the body and expose the ureteral area and the freed lower end of the ureter and the ureterovesical junction;

[0031] (c) Ligate the bladder tissue below the ureterovesical junction;

[0032] (d) The abdominal muscles and skin were closed by suturing with sutures.

[0033] The construction method of this invention, through specific construction method settings, can form an obstruction area in the bladder region of mice without causing damage to the ureter, so that the model construction success rate can reach 100% and the recanalization surgery success rate can reach more than 90%. It avoids the problems of ureteral damage caused by external compression of the ureter in existing technologies and reimplantation methods, and effectively solves the problem that recanalization failure or incomplete recanalization is easy to occur in existing technologies.

[0034] In one embodiment, ligating the bladder tissue below the ureterovesical junction includes clamping with a vascular clamp; ligating with a suture; or removing a section of silicone tube and then passing a suture through the silicone tube to ligate the bladder tissue.

[0035] The present invention does not specify the method of anesthesia for mice, and conventional anesthesia methods in the art can be used. In one embodiment, in step (a), the anesthesia is performed by intraperitoneal injection of 1.0% sodium pentobarbital at a dose of 50 mg / kg into the mice.

[0036] In one embodiment, in step (b), the incision length can be any length between 0.6 and 1.5 cm. Preferably, in one embodiment, the incision length is 0.8 cm.

[0037] In one embodiment, in step (b), the length of the ureter freed can be any length between 0.3 and 0.5 cm.

[0038] In one embodiment, in step (c), the length of the silicone tube can be any length between 2.5 and 3.5 mm, the inner diameter can be any value between 1.2 and 1.8 mm, and the outer diameter can be any value between 2.2 and 2.8 mm.

[0039] In one embodiment, in step (d), the sutures can all be 5-0 sutures.

[0040] In one embodiment, the construction method has a 100% success rate.

[0041] In one embodiment, the knot is tied during the ligation process to avoid the ureterovesical junction and the urethra.

[0042] Another embodiment of the present invention provides a mouse model of reversible ureteral obstruction in which the obstruction region is located in the bladder, constructed by the above construction method.

[0043] The technical solution of the present invention will be further described in detail below through specific embodiments.

[0044] Example

[0045] This embodiment describes a method for constructing a mouse model of reversible ureteral obstruction, which includes the following steps:

[0046] (a) Thirty adult male C57BL / 6 mice, 6-8 weeks old, weighing 20±2 grams, were purchased from Chongqing Medical University (Production License No.: ) and housed in the experimental animal center. The mice were acclimatized for 1 week before surgery and placed on a warm blanket for 24 hours after surgery to facilitate recovery. The mice were anesthetized by intraperitoneal injection of 1.0% sodium pentobarbital 50 mg / kg and then fixed in a supine position for skin preparation and disinfection.

[0047] (b) Make a midline incision in the lower abdomen of the mouse, with an incision length of 0.8 cm; remove the bladder from the body and expose the left ureteral region and free the lower end of the left ureter and the ureterovesical junction, wherein the free length of the left ureter is 0.3-0.5 cm;

[0048] (c) Clamp the bladder tissue at the ureterovesical junction with a vascular clamp; then take out a 3mm long silicone tube with an inner diameter of 1.5mm and an outer diameter of 2.5mm. Use 5-0 sutures to ligate the bladder tissue below the vascular clamp. During the ligation process, avoid the ureterovesical junction and the urethra at the knot. After tying the silicone tube, make the upper and lower walls close to each other, but do not make them too close together or leave a large gap.

[0049] (d) Two interrupted sutures were used to close the abdominal muscles and skin.

[0050] The mouse manipulation diagrams for each step in the above construction method are shown below. Figure 1 As shown, Figure 1 In this study, after anesthetizing mice, the mice were placed in a supine position. After skin preparation and disinfection, a midline incision was made in the lower abdomen, and the incision was made layer by layer to the abdominal cavity. The incision was about 0.8 cm long.

[0051] b. Locate the bladder and remove it from the body;

[0052] c. Expose the left ureteral region and free the lower end of the left ureter and the ureterovesical junction;

[0053] d. Use a vascular clamp to clamp the bladder tissue at the vesicoureteral junction (to facilitate knotting);

[0054] e. Use 5-0 silk suture to ligate the bladder tissue below the vascular clamp (Note 1: Avoid ligating the urethra or ureter at the knot).

[0055] f. Status after tubal ligation;

[0056] The following three diagrams illustrate the re-processing:

[0057] g: Upon recanalization, open the abdominal cavity, remove the bladder, and remove the ligation sutures;

[0058] h. Gently pull on the bladder tissue to loosen the adhesions on the inner wall of the bladder; cut open any areas that cannot be loosened.

[0059] i. The bladder incision was sutured.

[0060] Ultrasound observation and magnetic resonance imaging were used to determine whether the mouse model of reversible ureteral obstruction was successfully established.

[0061] Then, 7 days after the establishment of the above mouse model, a recanalization surgery was performed (the ligature was removed and the bladder rupture was sutured), and ultrasound observation was used to determine whether the recanalization surgery was successful.

[0062] The results of ultrasound observation are as follows Figure 2 As shown, Figure 2 UUO in the middle refers to postoperative treatment, and RUU refers to recanalization.

[0063] The success rate of constructing a mouse model of reversible ureteral obstruction and the success rate of recanalization surgery were statistically analyzed. The results are shown in Table 1.

[0064] Table 1

[0065] Group Success rate Recanalization surgery success rate Example 100％ 90％ External compression of the ureter 100％ 60％

[0066] As shown in Table 1:

[0067] The method described in this application can successfully construct a mouse model of plastic tube obstruction, and the recanalization surgery has a high success rate.

[0068] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be covered within the scope of the claims and specification of the present invention.

Claims

1. A method for constructing a mouse model of reversible ureteral obstruction, characterized in that, The construction method includes the following steps: (a) Anesthetize the mice, then fix them in a supine position and prepare and disinfect their skin; (b) Make an incision in the middle of the lower abdomen of the mouse, remove the bladder from the body and expose the ureteral area and the freed lower end of the ureter and the ureterovesical junction; (c) Ligate the bladder tissue below the ureterovesical junction; (d) The abdominal muscles and skin were closed by suturing with sutures.

2. The construction method according to claim 1, characterized in that, In step (c), ligating the bladder tissue below the ureterovesical junction includes clamping with a vascular clamp; ligating with a suture; or removing a section of silicone tube and then ligating the bladder tissue by passing a suture through the silicone tube.

3. The construction method according to claim 1, characterized in that, In step (b), the incision length is 0.6 to 1.5 cm.

4. The construction method according to claim 1, characterized in that, In step (b), the length of the ureter freed is 0.3 to 0.5 cm.

5. The construction method according to claim 1, characterized in that, During the ligation process, the knot should be tied away from the ureterovesical junction and the urethra.

6. The construction method according to claim 1, characterized in that, The construction method described above has a 100% success rate.

7. A mouse model of reversible ureteral obstruction with the obstruction region located in the bladder, constructed by any of the construction methods described in claims 1 to 6.

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