Verification method of rat neuropathic pain model

Abstract

The invention discloses a verification method of a rat neuropathic pain model, and relates to the field of clinical medicine. Comprising the following steps: animal preparation: selecting healthy adult rats, and putting the rats in a laboratory environment for adaptive feeding; model construction: constructing a sciatic nerve branch selective injury model; the pain behavioral test comprises a foot sole hot stabbing pain experiment, a mechanical pain sense test and a cold pain sense test; data analysis: performing data analysis according to a behavioral test result; and model verification: verifying whether the construction of the model is successful or not by observing the behavioral performance of the rat, comparing the symptoms of the model rat with the symptoms of a human neuropathic pain patient, and evaluating the clinical similarity of the model. According to the method, the neuropathic pain model is successfully induced and verified through SNI model surgical operation and hot plate testing, and the method has high reliability and repeatability and provides powerful support for research of neuropathic pain.

Description

Technical Field

[0001] The invention relates to the field of clinical medicine, and in particular to a verification method for a rat neuropathic pain model. Background Art

[0002] Neuropathic pain is a painful condition caused by damage or disease of the nervous system, which has complex mechanisms and is difficult to treat.

[0003] Neuropathic pain can be caused by a variety of reasons, including: Physical injury: nerve damage caused by trauma, surgery, etc. Chemical injury: neurotoxicity from drugs or radiotherapy. Metabolic complex neuropathy: nerve damage caused by metabolic diseases such as diabetes. Other causes: including infection, poisoning, vascular disease, nutritional disorders, tumors, nerve compression, immunity, genetics, etc.

[0004] The clinical features of neuropathic pain mainly include: spontaneous pain: pain that occurs without any external stimulation. Hyperalgesia: excessive pain response to normal stimulation. Abnormal pain: such as touch pain, that is, stimulation that does not cause pain to normal people causes pain to patients. Abnormal sensation: such as numbness, itching, and tingling sensation.

[0005] In order to deeply study the pathological mechanisms of neuropathic pain and develop new treatments, it is crucial to establish a reliable animal model.

[0006] To this end, the present invention proposes a verification method for a rat neuropathic pain model Summary of the invention

[0007] The purpose of the present invention is to solve the shortcomings of the prior art and to propose a verification method for a rat neuropathic pain model.

[0008] In order to achieve the above object, the present invention adopts the following technical solutions:

[0009] A method for verifying a rat neuropathic pain model comprises the following steps:

[0010] S1: Animal preparation: healthy adult rats were selected and placed in a laboratory environment for adaptive breeding;

[0011] S2: Model construction, constructing a sciatic nerve branch selective injury model;

[0012] S3: pain behavior test, including plantar heat pain test, mechanical pain test, and cold pain test;

[0013] S4: Data analysis, data analysis based on behavioral test results;

[0014] S5: Model validation: verify whether the model is successfully constructed by observing the behavioral performance of rats, and compare the symptoms of model rats with those of human neuropathic pain patients to evaluate the clinical similarity of the model.

[0015] Preferably: in step S1, the weight of the rats is 200-300 grams, and the feeding time is one to two weeks.

[0016] Preferably: the step S2 comprises the following steps:

[0017] S21: After the rats were deeply anesthetized, the hair on the left side below the pelvis was shaved;

[0018] S22: Cut the skin and muscles of the hind limb to expose the sciatic nerve and its terminal branches;

[0019] S23: The peroneal and tibial nerves were separated from the fascia, ligated with No. 5.0 silk thread, and then transected distally, preserving the sural nerve;

[0020] S24: The wound is sutured, and the rat is then fed and allowed to recover for a certain period.

[0021] Preferably: in step S21, the anesthetic is sodium pentobarbital at a dosage of 45-50 mg / kg.

[0022] Preferably: in step S3, the plantar pain test comprises the following steps:

[0023] A1: Heat the hot plate and set the temperature to 52±0.5 degrees Celsius;

[0024] A2: Place the rats individually on the hot plate. When the rats show avoidance behavior, stop heating immediately and record the time. This time is the pain latency period, which reflects the thermal pain value of the rats.

[0025] A3: Repeat step A2 3-5 times, with each time interval at least 15 minutes, and take the average value as the result.

[0026] Preferably: in step S3, the mechanical pain test comprises the following steps:

[0027] B1: Using an electronic pressure analgesia instrument, the mechanical pressure and pressure duration when the rats showed avoidance behavior were measured;

[0028] B2: Repeat step B1 3-5 times, with a time interval of at least 15 minutes between each step, and take the average value as the result.

[0029] Preferably: in step S3, the cold pain test comprises the following steps:

[0030] C1: Place the rat on a constant temperature cold plate, set at 0-5°C;

[0031] C2: measuring the time when the rats showed avoidance behavior;

[0032] C3: Repeat step C2 3-5 times, with a time interval of at least 15 minutes between each step, and take the average value as the result.

[0033] Preferably: in the steps A2, B1 and C2, the sign of the rat's avoidance behavior is any one of the actions of the rat licking its feet, jumping and lifting its feet.

[0034] Preferably: the step S4 comprises the following steps:

[0035] S41: Record the pain latency, mechanical pain threshold and cold pain response time of each rat;

[0036] S42: Calculate the mean and standard deviation of all rats;

[0037] S43: Use t-test or analysis of variance to compare whether there are significant differences in pain latency, mechanical pain threshold, and cold pain response time between different treatment groups;

[0038] S44: If the p-value is less than 0.05, it is statistically significant and the validation data is valid.

[0039] The beneficial effects of the present invention are:

[0040] 1. The present invention successfully induced and verified the neuropathic pain model through SNI model surgical operation and hot plate test. This method has high reliability and repeatability, providing strong support for the research of neuropathic pain. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 This is a flow chart of the verification method of a rat neuropathic pain model proposed in the present invention. DETAILED DESCRIPTION

[0042] The technical solution of the present invention is further described in detail below in conjunction with specific implementation methods.

[0043] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", and "set" should be understood in a broad sense, for example, they can be fixedly connected or set, or detachably connected or set, or integrally connected or set. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0044] Embodiment 1:

[0045] A method for verifying a rat neuropathic pain model comprises the following steps:

[0046] S1: Animal preparation: healthy adult rats were selected and placed in a laboratory environment for adaptive breeding;

[0047] S2: Model construction, constructing a sciatic nerve branch selective injury model;

[0048] S3: pain behavior test, including plantar heat pain test, mechanical pain test, and cold pain test;

[0049] S4: Data analysis, data analysis based on behavioral test results;

[0050] S5: Model validation: verify whether the model is successfully constructed by observing the behavioral performance of rats, and compare the symptoms of model rats with those of human neuropathic pain patients to evaluate the clinical similarity of the model.

[0051] In the step S1, the weight of the rats is 200-300 grams, and the feeding time is one to two weeks.

[0052] Embodiment 2:

[0053] A method for verifying a rat neuropathic pain model comprises the following steps:

[0054] S1: Animal preparation: healthy adult rats were selected and placed in a laboratory environment for adaptive breeding;

[0055] S2: Model construction, constructing a sciatic nerve branch selective injury model;

[0056] S3: pain behavior test, including plantar heat pain test, mechanical pain test, and cold pain test;

[0057] S4: Data analysis, data analysis based on behavioral test results;

[0058] S5: Model validation: verify whether the model is successfully constructed by observing the behavioral performance of rats, and compare the symptoms of model rats with those of human neuropathic pain patients to evaluate the clinical similarity of the model.

[0059] In the step S1, the weight of the rats is 200-300 grams, and the feeding time is one to two weeks.

[0060] The step S2 comprises the following steps:

[0061] S21: After the rats were deeply anesthetized, the hair on the left side below the pelvis was shaved;

[0062] S22: Cut the skin and muscles of the hind limb to expose the sciatic nerve and its terminal branches;

[0063] S23: The peroneal and tibial nerves were separated from the fascia, ligated with No. 5.0 silk thread, and then transected distally, preserving the sural nerve;

[0064] S24: The wound is sutured, and the rat is then fed and allowed to recover for a certain period.

[0065] In step S21, sodium pentobarbital is used as the anesthetic at a dosage of 45-50 mg / kg.

[0066] Embodiment 3:

[0067] A method for verifying a rat neuropathic pain model comprises the following steps:

[0068] S1: Animal preparation: healthy adult rats were selected and placed in a laboratory environment for adaptive breeding;

[0069] S2: Model construction, constructing a sciatic nerve branch selective injury model;

[0070] S3: pain behavior test, including plantar heat pain test, mechanical pain test, and cold pain test;

[0071] S4: Data analysis, data analysis based on behavioral test results;

[0072] S5: Model validation: verify whether the model is successfully constructed by observing the behavioral performance of rats, and compare the symptoms of model rats with those of human neuropathic pain patients to evaluate the clinical similarity of the model.

[0073] In the step S1, the weight of the rats is 200-300 grams, and the feeding time is one to two weeks.

[0074] The step S2 comprises the following steps:

[0075] S21: After the rats were deeply anesthetized, the hair on the left side below the pelvis was shaved;

[0076] S22: Cut the skin and muscles of the hind limb to expose the sciatic nerve and its terminal branches;

[0077] S23: The peroneal and tibial nerves were separated from the fascia, ligated with No. 5.0 silk thread, and then transected distally, preserving the sural nerve;

[0078] S24: The wound is sutured, and the rat is then fed and allowed to recover for a certain period.

[0079] In step S21, sodium pentobarbital is used as the anesthetic at a dosage of 45-50 mg / kg.

[0080] In step S3, the plantar tingling test includes the following steps:

[0081] A1: Heat the hot plate and set the temperature to 52±0.5 degrees Celsius;

[0082] A2: Place the rats individually on the hot plate. When the rats show avoidance behavior, stop heating immediately and record the time. This time is the pain latency period, which reflects the thermal pain value of the rats.

[0083] A3: Repeat step A2 3-5 times, with each time interval at least 15 minutes, and take the average value as the result.

[0084] In step S3, the mechanical pain test includes the following steps:

[0085] B1: Using an electronic pressure analgesia instrument, the mechanical pressure and pressure duration when the rats showed avoidance behavior were measured;

[0086] B2: Repeat step B1 3-5 times, with a time interval of at least 15 minutes between each step, and take the average value as the result.

[0087] In step S3, the cold pain test includes the following steps:

[0088] C1: Place the rat on a constant temperature cold plate, set at 0-5°C;

[0089] C2: measuring the time when the rats showed avoidance behavior;

[0090] C3: Repeat step C2 3-5 times, with a time interval of at least 15 minutes between each step, and take the average value as the result.

[0091] In the steps A2, B1 and C2, the sign of the avoidance behavior of the rat is any one of licking the foot, jumping and lifting the foot.

[0092] Embodiment 4:

[0093] A method for verifying a rat neuropathic pain model comprises the following steps:

[0094] S1: Animal preparation: healthy adult rats were selected and placed in a laboratory environment for adaptive breeding;

[0095] S2: Model construction, constructing a sciatic nerve branch selective injury model;

[0096] S3: pain behavior test, including plantar heat pain test, mechanical pain test, and cold pain test;

[0097] S4: Data analysis, data analysis based on behavioral test results;

[0098] S5: Model validation: verify whether the model is successfully constructed by observing the behavioral performance of rats, and compare the symptoms of model rats with those of human neuropathic pain patients to evaluate the clinical similarity of the model.

[0099] In the step S1, the weight of the rats is 200-300 grams, and the feeding time is one to two weeks.

[0100] The step S2 comprises the following steps:

[0101] S21: After the rats were deeply anesthetized, the hair on the left side below the pelvis was shaved;

[0102] S22: Cut the skin and muscles of the hind limb to expose the sciatic nerve and its terminal branches;

[0103] S23: The peroneal and tibial nerves were separated from the fascia, ligated with No. 5.0 silk thread, and then transected distally, preserving the sural nerve;

[0104] S24: The wound is sutured, and the rat is then fed and allowed to recover for a certain period.

[0105] In step S21, sodium pentobarbital is used as the anesthetic at a dosage of 45-50 mg / kg.

[0106] In step S3, the plantar tingling test includes the following steps:

[0107] A1: Heat the hot plate and set the temperature to 52±0.5 degrees Celsius;

[0108] A2: Place the rats individually on the hot plate. When the rats show avoidance behavior, stop heating immediately and record the time. This time is the pain latency period, which reflects the thermal pain value of the rats.

[0109] A3: Repeat step A2 3-5 times, with each time interval at least 15 minutes, and take the average value as the result.

[0110] In step S3, the mechanical pain test includes the following steps:

[0111] B1: Using an electronic pressure analgesia instrument, the mechanical pressure and pressure duration when the rats showed avoidance behavior were measured;

[0112] B2: Repeat step B1 3-5 times, with a time interval of at least 15 minutes between each step, and take the average value as the result.

[0113] In step S3, the cold pain test includes the following steps:

[0114] C1: Place the rat on a constant temperature cold plate, set at 0-5°C;

[0115] C2: measuring the time when the rats showed avoidance behavior;

[0116] C3: Repeat step C2 3-5 times, with a time interval of at least 15 minutes between each step, and take the average value as the result.

[0117] In the steps A2, B1 and C2, the sign of the avoidance behavior of the rat is any one of licking the foot, jumping and lifting the foot.

[0118] The S4 step comprises the following steps:

[0119] S41: Record the pain latency, mechanical pain threshold and cold pain response time of each rat;

[0120] S42: Calculate the mean and standard deviation of all rats;

[0121] S43: Use t-test or analysis of variance to compare whether there are significant differences in pain latency, mechanical pain threshold, and cold pain response time between different treatment groups;

[0122] S44: If the p-value is less than 0.05, it is statistically significant and the validation data is valid.

[0123] experiment:

[0124] 60 adult rats were selected and divided into two groups, namely experimental group A and control group B. Step S2 was performed on the adult rats in experimental group A, and then step S3 was performed on experimental group A and control group B. The data obtained are as follows:

[0125]

[0126] As shown in the table, the pain latency of rats in the SNI model group was significantly lower than that in the normal control group (p<0.05), indicating that the SNI model successfully induced neuropathic pain in rats.

[0127] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A method for validating a rat neuropathic pain model, characterized in that: The following steps are involved: S1: Animal preparation: healthy adult rats were selected and placed in a laboratory environment for adaptive breeding; S2: Model construction, constructing a sciatic nerve branch selective injury model; S3: pain behavior test, including plantar heat pain test, mechanical pain test, and cold pain test; S4: Data analysis, data analysis based on behavioral test results; S5: Model validation: verify whether the model is successfully constructed by observing the behavioral performance of rats, and compare the symptoms of model rats with those of human neuropathic pain patients to evaluate the clinical similarity of the model.

2. The method for verifying a rat neuropathic pain model according to claim 1, characterized in that: In the step S1, the weight of the rats is 200-300 grams, and the feeding time is one to two weeks.

3. The method for verifying a rat neuropathic pain model according to claim 1, characterized in that: The step S2 comprises the following steps: S21: After the rats were deeply anesthetized, the hair on the left side below the pelvis was shaved; S22: Cut the skin and muscles of the hind limb to expose the sciatic nerve and its terminal branches; S23: The peroneal and tibial nerves were separated from the fascia, ligated with No. 5.0 silk thread, and then transected distally, preserving the sural nerve; S24: The wound is sutured, and the rat is then fed and allowed to recover for a certain period.

4. The method for verifying a rat neuropathic pain model according to claim 3, characterized in that: In step S21, sodium pentobarbital is used as the anesthetic at a dosage of 45-50 mg / kg.

5. The method for verifying a rat neuropathic pain model according to claim 1, characterized in that: In step S3, the plantar tingling test includes the following steps: A1: Heat the hot plate and set the temperature to 52±0.5 degrees Celsius; A2: Place the rats individually on the hot plate. When the rats show avoidance behavior, stop heating immediately and record the time. This time is the pain latency period, which reflects the thermal pain value of the rats. A3: Repeat step A2 3-5 times, with each time interval at least 15 minutes, and take the average value as the result.

6. The method for verifying a rat neuropathic pain model according to claim 5, characterized in that: In step S3, the mechanical pain test includes the following steps: B1: Using an electronic pressure analgesia instrument, the mechanical pressure and pressure duration when the rats showed avoidance behavior were measured; B2: Repeat step B1 3-5 times, with a time interval of at least 15 minutes between each step, and take the average value as the result.

7. The method for verifying a rat neuropathic pain model according to claim 6, characterized in that: In step S3, the cold pain test includes the following steps: C1: Place the rat on a constant temperature cold plate, set at 0-5°C; C2: measuring the time when the rats showed avoidance behavior; C3: Repeat step C2 3-5 times, with a time interval of at least 15 minutes between each step, and take the average value as the result.

8. The method for verifying a rat neuropathic pain model according to claim 7, characterized in that: In the steps A2, B1 and C2, the sign of the avoidance behavior of the rat is any one of licking the foot, jumping and lifting the foot.

9. The method for verifying a rat neuropathic pain model according to claim 1, characterized in that: The S4 step comprises the following steps: S41: Record the pain latency, mechanical pain threshold and cold pain response time of each rat; S42: Calculate the mean and standard deviation of all rats; S43: Use t-test or analysis of variance to compare whether there are significant differences in pain latency, mechanical pain threshold, and cold pain response time between different treatment groups; S44: If the p-value is less than 0.05, it is statistically significant and the validation data is valid.

Images