Research method of action mechanism of visual needle-knife in treatment of femoral head necrosis stasis pain

Abstract

The invention discloses a research method of an action mechanism of a visual needle-knife on treatment of femoral head necrosis stasis pain, and belongs to the technical field of traditional Chinese medicine needle-knife therapies, and the research method comprises the following steps: S1, establishing a femoral head necrosis animal model; s2, establishing a femoral head necrosis pain trigger point model; s3, positioning and marking the femoral head necrosis pain trigger points on the femoral head necrosis pain trigger point model; s4, grouping the caput femoris necrosis pain trigger point models marked in the step S3, and detecting an inflammatory factor expression level, a vasomotor factor expression level and a neurotransmitter content in animal serum of each group; s5, performing high-throughput screening; s6, performing low-throughput screening and network construction on the neurotransmitters to obtain a neurotransmitter-gene metabolism interaction network; and S7, in combination with the step S4, the step S5 and the step S6, determining the action mechanism of the visual needle-knife in treating the femoral head necrosis stasis pain.

Description

Technical Field

[0001] The present invention belongs to the field of traditional Chinese medicine acupuncture knife therapy, and specifically relates to a research method for the mechanism of action of a visualized acupuncture knife in treating femoral head necrosis and pain. Background Art

[0002] Osteonecrosis of the femoral head (ONFH) is a progressive and difficult orthopedic disease. Patients often experience joint dysfunction due to hip pain, with pain and stasis at the core of the disease's progression. Acupuncture has traditionally demonstrated excellent therapeutic effects for pain and stasis. Acupotomy, as an emerging treatment method, exemplifies the integration of acupuncture instruments in both Chinese and Western medicine. It is simple to learn and effective, and has been widely used in the clinical treatment of pain disorders, especially chronic pain. Acupotomy has the potential to release tissue, regulate inflammatory factors, modulate endogenous neurotransmitters, and repair soft tissue damage. While a previously established visual acupotomy system has shown satisfactory results in treating pain and stasis in ONFH, its mechanism of action remains unclear, limiting its clinical application. Summary of the Invention

[0003] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a method for studying the mechanism of action of a visual acupotomy in treating femoral head necrosis and pain.

[0004] Based on the existing research foundation that the interaction between pain trigger points and neurotransmitter-muscle fiber metabolic disorders is the key to the pathology of "stasis pain," this paper proposes the scientific hypothesis that visual acupotomy may achieve its therapeutic effect by relieving pain trigger points, regulating this interaction, and correcting circulatory disturbances. To verify this hypothesis, the present invention utilized an ONFH animal model to pinpoint the location of pain trigger points and evaluate their efficacy.

[0005] To achieve the above object, the technical solution adopted by the present invention is:

[0006] A method for studying the mechanism of action of a visual acupotomy in treating femoral head necrosis and pain, comprising the following steps:

[0007] S1. Establish an animal model of femoral head necrosis;

[0008] S2. Based on the femoral head necrosis animal model established in step S1, a femoral head necrosis pain trigger point model is established;

[0009] S3, locating and marking the femoral head necrosis pain trigger points on the femoral head necrosis pain trigger point model obtained in step S2;

[0010] S4. The femoral head necrosis pain trigger point models marked in step S3 are divided into a model control group, a traditional acupotomy group, and a visual acupotomy group. The expression levels of inflammatory factors, the expression levels of vasomotor factors, and the content of neurotransmitters in the serum of the animals in each group are tested before and after the intervention with traditional acupotomy and visual acupotomy, respectively, to obtain the test results.

[0011] S5. Using gene chips, high-throughput screening is performed on the model control group and the visual acupotomy group obtained in step S4 to obtain genes with differential therapeutic effects;

[0012] S6. Performing low-throughput neurotransmitter screening and network construction on the model control group and the visual acupotomy group obtained in step S4 to obtain a neurotransmitter-gene metabolism interaction network;

[0013] S7. Combining the detection results of step S4, the differential efficacy genes of step S5 and the neurotransmitter-gene metabolism interaction network of step S6, the potential core targets of visual acupotomy in treating pain and bruise of femoral head necrosis are obtained, that is, the mechanism of action of visual acupotomy in treating pain and bruise of femoral head necrosis is clarified.

[0014] Preferably, step S1 comprises the following steps:

[0015] S11. After anesthetizing the beagle dog, one forelimb was depilated and routinely disinfected. A Kirschner wire was used to fix the carpometacarpal joint of the dog through an anterior incision. The end of the Kirschner wire was buried subcutaneously and sutured. The dog recovered for 2 weeks to obtain a three-legged beagle model.

[0016] S12. Depilate one side of the buttocks of the three-legged beagle model obtained in step S1. After routine disinfection, drill a 3.5 mm diameter bone tunnel from the greater trochanter of the femur to the femoral head, 2 mm below the surface of the articular cartilage. After confirming the satisfactory position through fluoroscopy, perfuse 0.5 MPa liquid nitrogen into the femoral head and freeze it for 2 minutes, then rewarm it for 2 minutes. Repeat the freezing and rewarming procedure 4 times for a total of 10 minutes. Close the wound and allow it to recover for 14 days to obtain an animal model of femoral head necrosis.

[0017] Preferably, step S2 comprises the following steps:

[0018] The femoral head necrosis animal model established in step S1 was subjected to treadmill eccentric exercise once a week, with the treadmill tilt angle of 16° and the speed gradually increased to 12 km / h for 60 minutes. The animals were fed normally for the remaining 6 days without eccentric exercise. This lasted for 8 weeks, and then recovered. During the recovery period, the animals were fed normally without any experimental intervention to obtain a femoral head necrosis pain trigger point model.

[0019] Preferably, step S3 comprises the following steps:

[0020] The femoral head necrosis pain trigger point model obtained in step S2 is anesthetized, and the femoral head necrosis pain trigger point model is placed in a supine position with the limbs fixed. The femoral intertrochanteric crest, rectus femoris, sartorius muscle, gluteus medius, and adductor muscle origin and insertion points are touched in turn. If an enlarged nodule is touched, it is determined to be the femoral head necrosis pain trigger point and marked.

[0021] Preferably, in step S6, the neurotransmitter-gene metabolism interaction network is constructed by constructing gene-metabolite through the metascape plug-in of cytoscape, and constructing the protein interaction network of peptide neurotransmitters through string.

[0022] Preferably, in step S7, the potential core targets are ACHE, PNMT and MAOB genes.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] The present invention uses an ONFH canine model to clearly locate pain trigger points and evaluate their efficacy. Furthermore, targeting pathological links such as electrophysiological changes and pain sensitivity changes at trigger points, joint tissue circulation, and motor function changes, the present invention uses electrophysiology, molecular biology, histology, gene chips, low-throughput screening of neurotransmitters, agonist / antagonist application, and behavioral techniques to discover that acupuncture can significantly improve the pain sensitivity threshold and discharge phenomenon of trigger points in experimental animals, and its efficacy has been verified in clinical studies. Further studies have found that significant changes occur in vasomotor factors in the venous blood of beagle dogs: the levels of ET-1 and NA, which have a vasoconstrictive effect, are significantly reduced, while the levels of NO, HIS, and CGRP, which dilate blood vessels, are significantly increased. This indicates that acupuncture can regulate the expression of vasomotor factors, promote the repair of femoral head blood supply, and achieve the purpose of treating ONFH pain. Objective: The experiment further used high-throughput screening of differential genes combined with low-throughput screening of neurotransmitter detection and gene-metabolite network construction analysis to find that ONFH has a complex abnormal association network of energy metabolism, muscle metabolism, microcirculation, pain synaptic transmission, inflammation and oxidative stress. The intersection analysis of the metabolic network and differential genes after low-throughput neurotransmitter screening and experimental verification found that ACHE, PNMT, and MAOB are the core targets of visual acupotomy for the treatment of ONFH "stasis pain", indicating that correcting the metabolic imbalance of acetylcholine, norepinephrine, vasoactive amines, and neuroactive amines at the pain trigger point and then improving microcirculation is one of the main mechanisms of action of visual acupotomy for the treatment of ONFH "stasis pain"; this invention enriches the molecular mechanism of acupotomy for the treatment of ONFH "stasis pain" and provides a reference for clinical promotion and further development of visual acupotomy robots. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Create an operating diagram for the three-legged beagle model;

[0026] Figure 2 Diagram for Beagles to adapt to three-legged walking;

[0027] Figure 3 To establish an operational diagram for the ONFH pain trigger point model;

[0028] Figure 4 This is a comparison of electrophysiological testing between the sham operation group and the model group;

[0029] Figure 5 This is an image of the femoral head of a beagle dog;

[0030] in, Figure 5 A and Figure 5 C are CT and MRI images of normal femoral head; Figure 5 B and Figure 5 D are CT and MRI images of the necrotic femoral head;

[0031] Figure 6 This is a gross photo of the femoral head of a beagle;

[0032] in, Figure 6 A. Figure 6 B and Figure 6 C is the normal femoral head; Figure 6 D. Figure 6 E and Figure 6 F is necrotic femoral head;

[0033] Figure 7 HE-stained sections of the hip joint surrounding tissues in each group. DETAILED DESCRIPTION

[0034] The following is a diagram of the embodiment of the present invention. Figures 1 to 7 The technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0035] The embodiment of the present invention provides a method for studying the mechanism of action of a visual acupotomy in treating femoral head necrosis and pain, comprising the following steps:

[0036] S1. Establish an animal model of femoral head necrosis, which specifically includes the following steps:

[0037] S11. After anesthetizing the beagle dog, one forelimb was depilated and routinely disinfected. A Kirschner wire was used to fix the carpometacarpal joint of the dog through an anterior incision. The end of the Kirschner wire was buried subcutaneously and sutured. The dog recovered for 2 weeks to obtain a three-legged beagle model.

[0038] S12. Depilate one side of the buttocks of the three-legged beagle model obtained in step S1. After routine disinfection, drill a 3.5 mm diameter bone tunnel from the greater trochanter of the femur to the femoral head, 2 mm below the surface of the articular cartilage. After confirming the satisfactory position through fluoroscopy, perfuse 0.5 MPa liquid nitrogen into the femoral head and freeze it for 2 minutes, then rewarm it for 2 minutes. Repeat the freezing and rewarming procedure 4 times for a total of 10 minutes. Close the wound and allow it to recover for 14 days to obtain an animal model of femoral head necrosis.

[0039] S2. Based on the femoral head necrosis animal model established in step S1, a femoral head necrosis pain trigger point model is established, which specifically comprises the following steps:

[0040] The femoral head necrosis animal model established in step S1 was subjected to treadmill eccentric exercise once a week, with the treadmill tilt angle of 16° and the speed gradually increased to 12 km / h for 60 minutes. The animals were fed normally for the remaining 6 days without eccentric exercise for 8 weeks, followed by a recovery week during which they were fed normally without any experimental intervention to obtain a femoral head necrosis pain trigger point model.

[0041] S3, locating and marking the femoral head necrosis pain trigger points on the femoral head necrosis pain trigger point model obtained in step S2, specifically comprising the following steps:

[0042] The femoral head necrosis pain trigger point model obtained in step S2 is anesthetized, and the femoral head necrosis pain trigger point model is placed in a supine position with the limbs fixed. The femoral intertrochanteric crest, rectus femoris, sartorius muscle, gluteus medius, and adductor muscle origin and insertion points are touched in turn. If an enlarged nodule is touched, it is determined to be the femoral head necrosis pain trigger point and marked.

[0043] S4. The femoral head necrosis pain trigger point models marked in step S3 are divided into a model control group, a traditional acupotomy group, and a visual acupotomy group. The expression levels of inflammatory factors, the expression levels of vasomotor factors, and the content of neurotransmitters in the serum of the animals in each group are tested before and after the intervention with traditional acupotomy and visual acupotomy, respectively, to obtain the test results.

[0044] S5. Using gene chips, high-throughput screening is performed on the model control group and the visual acupotomy group obtained in step S4 to obtain genes with differential therapeutic effects;

[0045] S6. Perform low-throughput neurotransmitter screening and network construction on the model control group and the visual acupotomy group obtained in step S4 to obtain a neurotransmitter-gene metabolism interaction network; the neurotransmitter-gene metabolism interaction network is constructed by constructing gene-metabolites through the metascape plug-in of cytoscape, and the protein interaction network of peptide neurotransmitters is constructed through string.

[0046] S7. Combining the detection results of step S4, the differential efficacy genes of step S5 and the neurotransmitter-gene metabolism interaction network of step S6, the potential core targets of visual acupotomy in the treatment of femoral head necrosis and pain are obtained. The potential core targets are ACHE, PNMT and MAOB genes, which clarifies the mechanism of action of visual acupotomy in the treatment of femoral head necrosis and pain.

[0047] The following is a detailed description of the research method of the mechanism of action of the visual acupotomy in treating femoral head necrosis and pain provided by the embodiment of the present invention.

[0048] 1. Localization and Verification of Pain Trigger Points in ONFH Animal Models

[0049] 1. Establishment of ONFH Animal Model

[0050] (1) Establishment of the three-legged beagle model:

[0051] The modeling period lasted for 2 weeks. In the present invention, 8 healthy adult male beagle dogs were randomly divided into a sham operation (Sham) group (4 dogs) and a model group (4 dogs). The two groups of beagle dogs were anesthetized with 10g / L sodium pentobarbital (30mg / kg) intravenously. After that, one side of the dog's forelimb was randomly selected for hair removal. After routine disinfection and draping, a small incision was made on the front side. Two Kirschner wires with a diameter of 2.0mm were used to fix the dog's carpometacarpal joint. The ends of the Kirschner wires were buried subcutaneously and sutured. The operation was as follows: Figure 1 After 2 weeks, the dogs adapted to three-legged walking and the three-legged beagle model was established. Figure 2 shown.

[0052] (2) Establishment of ONFH model

[0053] Four beagle dogs in the model group were anesthetized and the buttocks of one tripod were randomly selected for hair removal. Following routine disinfection and draping, a 3.5 mm diameter bone tunnel was drilled from the greater trochanter to the femoral head under C-arm X-ray guidance, reaching 2 mm below the articular cartilage surface. After satisfactory positioning was confirmed by fluoroscopy, 0.5 MPa liquid nitrogen was infused into the femoral head through a 3 mm arthrocentesis needle and frozen for 2 minutes, followed by 2 minutes of thawing. This freeze-thaw cycle was repeated four times for a total of 10 minutes. The probe was removed, the bone trough sealed with bone wax, and the wound closed. Sutures were removed 14 days postoperatively, and the tripod ONFH animal model was established by liquid nitrogen freezing. All experimental animals were housed in large enough cages to allow for free movement. Beagle dogs in the sham group underwent the same procedures as the model group after the proximal femur was exposed without further treatment.

[0054] 2. Establishment and positioning of ONFH pain trigger point model

[0055] (1) Modeling and recovery periods

[0056] The modeling period lasted for 8 weeks. After the sutures were removed, the model group and the sham group underwent eccentric treadmill exercise once a week. The treadmill was tilted at 16° and the speed was gradually increased to 12 km / h for 60 minutes. Figure 3 The remaining 6 days were followed by normal feeding, with no other experimental interventions. Recovery period: Both groups were followed by normal feeding, with no experimental interventions.

[0057] (2) Localization of ONFH pain trigger points

[0058] After the recovery period of the beagle dogs in the sham operation group and the model group, they were anesthetized by intravenous injection of sodium pentobarbital (30 mg / kg) at a concentration of 10 g / L. After successful anesthesia, the beagle dogs were placed in a supine position with their limbs fixed. An experienced clinician used the pads of his thumb or index finger to touch the intertrochanteric crest of the femur, the rectus femoris, sartorius, gluteus medius, and the origin and insertion of the adductor muscles in turn. Once an enlarged tubercle was touched, it was identified as a possible trigger point and marked. Subsequently, two fine needle electrodes were inserted into the possible trigger point (the reference electrode was located on the right tibia), and the two needles were separated by 1 to 2 cm to record the resting electromyographic signal for 10 minutes and save it for analysis. The sham operation group recorded the skeletal muscle corresponding to the model group. Electrophysiological examination observed spontaneous electromyographic signals at the enlarged tubercle around the hip joint on the modeling side of the model group, while there were no spontaneous electromyographic signals in the sham operation group. Figure 4 shown.

[0059] 2. A histological and behavioral study of visual acupotomy on trigger point pain in femoral head necrosis beagle dogs

[0060] 1. Grouping of experimental animals

[0061] Eighteen healthy adult male beagle dogs in the ONFH pain trigger point model established above were randomly divided into a model group, a visual acupotomy treatment group, and a traditional acupotomy treatment group, with 6 dogs in each group. The 6 healthy adult male beagle dogs in the sham group served as the control group.

[0062] 2. Intervention methods

[0063] The animals in the sham and model groups were housed and moved normally. The traditional acupotomy group received traditional acupotomy treatment at week 13, once a week for three sessions per course for a total of three weeks. The visual acupotomy group received visual acupotomy treatment at week 13, once a week for three sessions per course for a total of three weeks.

[0064] Traditional acupuncture knife operation method: Traditional acupuncture knife treatment is mainly performed on the pain trigger points in the adductor femoris, intertrochanteric crest, rectus femoris, sartorius muscle, gluteus medius, etc. The specific operation is as follows:

[0065] In the traditional acupotomy group, beagle dogs were placed in the supine position. Electrophysiological testing was performed to detect trigger points in the adductor vastus, intertrochanteric crest, rectus femoris, sartorius, and gluteus medius muscles, which were marked with gentian violet. Routine disinfection was performed, and sterile drapes were laid. The surgeon, wearing sterile gloves, held the acupotomy with their right hand and palpated the skin at the insertion point with their left hand. Using the four-step acupotomy technique, the trigger points in the adductor vastus, intertrochanteric crest, rectus femoris, sartorius, and gluteus medius muscles were incised, separated, and released. The surgeon removed the needle after the sensation of a string disappeared. Pressure was then applied to stop bleeding for 3 minutes. After observing for signs of active bleeding, a sterile gauze dressing was applied, and the surgical area was kept dry.

[0066] Visual acupotomy operation method: At week 13, for the beagle dogs in the visual acupotomy group, visual acupotomy treatment was performed under the guidance of the visual acupotomy system, following the path prompted by the visual acupotomy system to directly reach the lesion area.

[0067] 3. Obtain materials

[0068] After the experimental beagle dogs were killed by air embolism, the tissues around the affected hip joint were quickly peeled off and repeatedly rinsed with 0.9% saline on ice. Joint tissues of 3×3×0.3 cm in size were taken and fixed in 4% paraformaldehyde solution for 24 hours.

[0069] 4. Behavioral Observation of Beagles in Each Group

[0070] The same experimenter observed the general activities, dietary changes, mental state, posture, barking and hip joint activities of the beagle dogs in each group before the experiment, at the 12th week (after the modeling), and at the 15th week (after the treatment).

[0071] 5. Histomorphological observation of femoral head in beagle dogs

[0072] HE staining was used to observe the tissues around the affected hip joint of beagle dogs. The steps for making HE-stained paraffin sections were as follows: the tissues were removed from the fixative, the target tissues were trimmed, and the tissues were dehydrated in a dehydrator according to gradient alcohol. The dehydrated and transparent tissues were immersed in wax and then embedded in an embedding machine. The embedded wax slices were placed on a paraffin microtome for sectioning. The slices were then repeatedly dewaxed in xylene and then dewaxed in different concentrations of alcohol. Finally, they were rinsed in distilled water, and the cell nuclei were stained with hematoxylin, the cytoplasm was stained with eosin, the sections were dehydrated and transparent, and the sections were sealed with neutral gum. The pathological morphology of the tissues around the hip joint of each group was observed under an optical microscope.

[0073] 6. Experimental Results

[0074] (1) Imaging results of femoral head in beagle dogs

[0075] like Figure 5As shown, hip CT scans in the model group showed a situating hip joint with no significant joint space narrowing. The femoral head articular surface was smooth, the trabecular structure was unclear, and a low-density area was visible within the head. No hip abnormalities were observed in the sham group. Hip MRI scans in the model group showed intact hip joint morphology, with a double-line sign within the head, mixed signals, bone marrow edema, and joint effusion.

[0076] (2) Behavioral observation results of beagles

[0077] Before the experiment began, beagles in all groups were active, sensitive to sound and light stimuli, and ate normally. They slept normally and were obedient. They barked normally, moved normally and calmly, and their limbs were coordinated and flexible during movement. During passive hip joint movement, they were emotionally stable, showed no resistance, and showed no noticeable pain.

[0078] At the end of the modeling process, the beagle dogs in the sham group showed no significant changes compared to before the experiment. They were active, ate normally, moved their limbs in coordinated fashion, offered no resistance to passive hip movement, and showed no noticeable pain. However, the beagle dogs in the model, traditional acupotomy, and visual acupotomy groups showed lethargy, decreased activity, and dilated pupils. They were highly alert, restless, and had difficulty sleeping while trying to protect their wounds. They howled when touched and exhibited a certain aggressive tendency. They were unable to push off the ground with the affected limb and had difficulty walking. Passive hip movement offered significant resistance, some with joint swelling, and significant pain.

[0079] After treatment, beagles in the sham group showed no significant changes. They were active, had a normal diet, and had coordinated limb movements. They showed no resistance to passive hip movement and no significant pain response. Beagles in the model group showed persistent lethargy, decreased activity, dilated pupils, and a state of high alert. They attempted to protect their wounds and were restless, unable to sleep, howled when touched, and exhibited aggressive tendencies. They were unable to push off the affected limb and had difficulty walking. Passive hip movement showed significant resistance, and some were accompanied by joint swelling and significant pain. Beagles in the traditional acupotomy group were slightly more active, with increased activity and diet compared to pre-treatment levels. Pupils were normal. They were compliant, easily asleep, and intermittently howled. They could push off the affected limb and walk for short periods. Most beagles showed no resistance to passive hip movement, although a small number still showed resistance and decreased pain response. The beagles in the visual acupotomy group were active, with slightly less activity than those in the sham group. They ate more and had normal pupils. They were compliant, slept normally, did not howl, and pushed off the ground with their affected limbs for a long time and walked. All beagles showed no obvious resistance to passive movement of the affected hip joint and no obvious pain response.

[0080] By observing the general activities, dietary changes, mental state, posture, barking and hip joint activities of the four groups of beagle dogs before the start of the experiment, at the end of modeling and after the end of treatment, the results showed that the beagle dogs in the sham group were generally normal; the beagle dogs in the model group showed mental fatigue, high alert state, and certain aggressiveness, and their affected limbs could not push off the ground, and they had difficulty walking and had obvious pain reactions; the beagle dogs in the traditional acupotomy group and the visual acupotomy group showed the same performance as the model group at the end of modeling, and their conditions improved after intervention treatment, and their performance tended to be the same as that of the sham group, and the visual acupotomy group performed better than the traditional acupotomy group.

[0081] (3) Histomorphological results of femoral head in beagle dogs

[0082] Grossly, the surface of the femoral head of a normal beagle dog is smooth and complete, while the surface of the necrotic femoral head is rough and incomplete, with obvious collapse. Figure 6 The HE-stained sections of the tissues around the affected hip joint of the beagle dog were observed under a light microscope. Figure 7 As shown, through Figure 7 It can be seen that the cytoplasm of the sham group was evenly stained, and the cell nuclei were neatly arranged; the muscle fibers were tightly arranged, with small gaps, no muscle fiber rupture, abundant blood vessels, no obvious inflammatory cell infiltration, and no cell edema. The cell nuclei of the model group were disorderly arranged; the muscle fibers were loose, with large gaps, blurred muscle striations, accompanied by muscle fiber rupture; and local connective tissue hyperplasia. The cell nuclei of the traditional acupuncture group were slightly disordered; muscle fiber gaps were visible, and local muscle striations were blurred; and the connective tissue was slightly hyperplastic. The cells of the visualized acupuncture group were evenly stained, the cell nuclei were relatively neatly arranged; the muscle fibers were tightly arranged; the blood vessels were abundant, and no obvious connective tissue hyperplasia was observed.

[0083] 3. The main mechanism of action of visual acupotomy in improving the pain and stasis of ONFH model animals

[0084] 1. Study on the effect of visual acupotomy on inflammatory factors in beagle dogs with femoral head necrosis pain trigger point model

[0085] In the present invention, the expression levels of inflammatory factors in beagle dog serum were detected by ELISA method, and the effects of different intervention methods on pain in beagle dogs with ONFH pain trigger point model were compared microscopically, thereby evaluating the recovery effect of visual acupotomy on local soft tissue in beagle dogs with ONFH pain trigger point model.

[0086] (1) Comparison of inflammatory factor levels in each group of samples before treatment

[0087] Compared with the sham group, the IL-1β, IL-6, TNF-α and capsaicin levels in the model group, traditional acupotomy group and visual acupotomy group were significantly increased (P<0.05); compared with the model group, there were no significant differences in the IL-1β, IL-6, TNF-α and capsaicin levels in the traditional acupotomy group and visual acupotomy group (P>0.05); compared with the traditional acupotomy group, there were no significant differences in the IL-1β, IL-6, TNF-α and capsaicin levels in the visual group (P>0.05), see Table 1 below.

[0088] Table 1 Comparison of inflammatory factor levels in samples of each group before treatment

[0089] Group Number of dogs IL-1β (pg / mL) IL-6 (pg / mL) TNF-α (ng / mL) Capsaicin (ppm) Visualization Group 6 <![CDATA[92.19±10.03 a ]]> <![CDATA[22.89±4.26 a ]]> <![CDATA[0.43±0.07 a ]]> <![CDATA[1.09±0.10 a ]]> Traditional Group 6 <![CDATA[97.40±15.93 a ]]> <![CDATA[26.85±6.00 a ]]> <![CDATA[0.50±0.06 a ]]> <![CDATA[1.10±0.05 a ]]> Model Group 6 <![CDATA[89.89±25.87 a ]]> <![CDATA[27.02±7.73 a ]]> <![CDATA[0.44±0.08 a ]]> <![CDATA[1.06±0.07 a ]]> Sham Group 6 12.56±1.70 1.83±0.54 0.21±0.02 0.48±0.04

[0090] Note: Compared with the Sham group, a P<0.05; compared with the model group, b P<0.05; compared with the traditional group, c P < 0.05;

[0091] (2) Comparison of serum inflammatory factor levels before and after visual acupotomy treatment

[0092] Compared with before treatment, the levels of IL-1β, IL-6, TNF-α and Capsaicin at 7 days and 21 days after treatment were statistically significant (P < 0.05); compared with 7 days after treatment, the levels of L-1β, IL-6, TNF-α and Capsaicin at 21 days after treatment were statistically significant (P < 0.05). The results are shown in Table 2 below.

[0093] Table 2 Comparison of serum inflammatory factor levels before and after visual acupotomy treatment

[0094] Inflammatory factors Before treatment After treatment 7d After treatment 21d IL-1β (pg / mL) 92.19±10.03 <![CDATA[61.99±3.84 * ]]> <![CDATA[40.78±5.37 *# ]]> 83.732 0.000 IL-6 (pg / mL) 22.89±4.26 <![CDATA[7.54±1.80 * ]]> <![CDATA[3.80±0.25 *# ]]> 84.127 0.000 TNF-α (ng / mL) 0.43±0.07 <![CDATA[0.30±0.02 * ]]> <![CDATA[0.23±0.04 *# ]]> 53.059 0.000 Capsaicin (ppm) 1.09±0.10 <![CDATA[0.85±0.04 * ]]> <![CDATA[0.66±0.04 *# ]]> 155.692 0.000

[0095] Note: Compared with before treatment, * P<0.05; compared with 7 days after treatment, # P < 0.05;

[0096] (3) Comparison of serum inflammatory factor levels before and after traditional acupuncture treatment

[0097] Compared with those before treatment, the IL-1β, IL-6 and Capsaicin levels at 7 d and 21 d after treatment were significantly different (P < 0.05), the TNF-α level at 7 d after treatment was not significantly different (P > 0.05), and the TNF-α level at 21 d after treatment was significantly different (P < 0.05); compared with those at 7 d after treatment, the L-1β, IL-6 and Capsaicin levels at 21 d after treatment were significantly different (P < 0.05), and the TNF-α level at 21 d after treatment was not statistically significant (P > 0.05). The results are shown in Table 3 below.

[0098] Table 3 Comparison of serum inflammatory factor levels before and after traditional acupuncture treatment

[0099]

[0100]

[0101] Note: Compared with before treatment, * P<0.05; compared with 7 days after treatment, # P < 0.05;

[0102] (4) Comparison of inflammatory factor levels between the visual acupotomy group and the traditional acupotomy group after treatment

[0103] After treatment, the two groups showed that the levels of IL-1β, IL-6 and TNF-α in the visualization group were lower than those in the traditional group, and the differences were statistically significant (P < 0.05). There was no significant difference in the capsaicin content between the two groups after treatment (P > 0.05). The results are shown in Table 4 below.

[0104] Table 4 Comparison of inflammatory factor levels between the visualization group and the traditional group after treatment

[0105] Group Number of dogs IL-1β (pg / mL) IL-6 (pg / mL) TNF-α (ng / mL) Capsaicin (ppm) Visualization Group 6 40.78±5.37 3.80±0.25 0.23±0.04 0.66±0.04 Traditional Group 6 49.16±3.96 5.01±1.01 0.34±0.05 0.70±0.05 P-value 0.023 0.032 0.004 0.123

[0106] 2. Study on the effect of visual acupotomy on vasomotor factors in beagle dogs with femoral head necrosis pain trigger point model

[0107] In the present invention, the expression levels of vasomotor factors in beagle dog serum were detected by ELISA method, and the effects of different intervention methods on pain in beagle dogs with ONFH pain trigger point model were compared microscopically, thereby evaluating the recovery effect of visual acupotomy on local soft tissue in beagle dogs with ONFH pain trigger point model.

[0108] (1) Comparison of vasomotor factor levels in each group of samples before treatment

[0109] Compared with the Sham group, the ET-1 and NA levels of the model group, traditional acupotomy group and visual acupotomy group were significantly increased (P<0.05), and the HIS, NO and CGRP levels were significantly decreased (P<0.05); compared with the model group, there were no significant differences in the ET-1, HIS, NA, NO and CGRP levels of the traditional acupotomy group and visual acupotomy group (P>0.05); compared with the traditional acupotomy group, there were no significant differences in the ET-1, HIS, NA, NO and CGRP levels of the visual acupotomy group (P>0.05). The results are shown in Table 5 below.

[0110] Table 5 Comparison of vasomotor factor levels in each group of samples before treatment

[0111]

[0112] Note: Compared with the Sham group, a P<0.05; compared with the model group, bP<0.05; compared with the traditional group, c P < 0.05;

[0113] (2) Comparison of serum vasomotor factors before and after visual acupotomy treatment

[0114] Compared with those before treatment, the differences in ET-1, HIS, NA, NO and CGRP levels at 7 days and 21 days after treatment were statistically significant (P < 0.05); compared with those at 7 days after treatment, the differences in ET-1, HIS, NA, NO and CGRP levels at 21 days after treatment were statistically significant (P < 0.05). The results are shown in Table 6 below.

[0115] Table 6 Comparison of serum vasomotor factor levels before and after visual acupotomy treatment

[0116] Vasomotor factors Before treatment After treatment 7d After treatment 21d ET-1 (pg / mL) 34.47±12.61 <![CDATA[20.04±3.51 * ]]> <![CDATA[10.81±0.89 *# ]]> 19.134 0.007 HIS (ng / mL) 231.98±39.26 <![CDATA[365.35±39.39 * ]]> <![CDATA[430.42±28.03 *# ]]> 110.384 0.000 NA (ng / mL) 41.51±5.11 <![CDATA[19.51±4.26 * ]]> <![CDATA[13.91±1.08 *# ]]> 109.577 0.000 NO (μmol / L) 6.21±0.95 <![CDATA[8.62±0.73 * ]]> <![CDATA[14.03±1.99 *# ]]> 41.866 0.000 CGRP1 (pg / mL) 103.33±5.84 <![CDATA[131.75±13.54 * ]]> <![CDATA[162.80±11.79 *# ]]> 81.519 0.000

[0117] Note: Compared with before treatment, * P<0.05; compared with 7 days after treatment, # P < 0.05;

[0118] (3) Comparison of serum vasomotor factors levels before and after traditional acupuncture treatment

[0119] Compared with those before treatment, the differences in ET-1, HIS, NA, NO and CGRP levels at 7 days and 21 days after treatment were statistically significant (P < 0.05); compared with those at 7 days after treatment, the differences in ET-1, HIS, NA, NO and CGRP levels at 21 days after treatment were statistically significant (P < 0.05). The results are shown in Table 7 below.

[0120] Table 7 Comparison of serum vasomotor factors levels before and after traditional acupuncture treatment

[0121] Vasomotor factors Before treatment After treatment 7d After treatment 21d ET-1 (pg / mL) 40.48±14.72 <![CDATA[24.21±2.55 * ]]> <![CDATA[15.83±2.72 *# ]]> 14.613 0.017 HIS (ng / mL) 246.96±34.40 <![CDATA[350.37±16.55 * ]]> <![CDATA[391.20±6.349 *# ]]> 130.341 0.000 NA (ng / mL) 41.49±9.91 <![CDATA[23.34±4.63 * ]]> <![CDATA[17.42±1.80 *# ]]> 36.236 0.000 NO (μmol / L) 7.08±1.01 <![CDATA[8.97±0.51 * ]]> <![CDATA[11.26±0.54 *# ]]> 77.507 0.000 CGRP1 (pg / mL) 105.06±5.99 <![CDATA[124.28±11.41 * ]]> <![CDATA[148.23±4.15 *# ]]> 101.312 0.000

[0122] Note: Compared with before treatment, * P<0.05; compared with 7 days after treatment, # P < 0.05;

[0123] (4) Comparison of vasomotor factors levels between the visual acupotomy group and the traditional acupotomy group after treatment

[0124] After treatment, the two groups showed that the ET-1 and NA levels in the visual acupotomy group were lower than those in the traditional group, while the NO, HIS and CGRP levels were higher than those in the traditional group. The differences were statistically significant (P < 0.05). The results are shown in Table 8 below.

[0125] Table 8 Comparison of vasomotor factors levels between the visualization group and the traditional group after treatment

[0126] Group Number of dogs ET-1 (pg / mL) HIS (ng / mL) NA (ng / mL) NO (μmol / L) CGRP (pg / mL) Visualization Group 6 10.81±0.89 430.42±28.03 13.91±1.08 14.03±1.99 162.80±11.79 Traditional Group 6 15.83±2.72 391.20±6.349 17.42±1.80 11.26±0.54 148.23±4.15 P-value 0.002 0.007 0.002 0.034 0.017

[0127] 3. Study on the effect of visual acupotomy on neurotransmitters in beagle dogs with femoral head necrosis pain trigger point model

[0128] (1) Comparison of neurotransmitter levels in samples from each group before treatment

[0129] Compared with the Sham group, the levels of 5-HT, SP, PGF2α, TK, BK, NPY and CGRP1 in the model group, traditional acupotomy group and visual acupotomy group were significantly increased (P<0.05); compared with the model group, there were no significant differences in the levels of 5-HT, SP, PGF2α, TK, BK, NPY and CGRP1 in the traditional acupotomy group and visual acupotomy group (P>0.05); compared with the traditional acupotomy group, there were no significant differences in the levels of 5-HT, SP, PGF2α, TK, BK, NPY and CGRP1 in the visual acupotomy group (P>0.05). The results are shown in Table 9 below.

[0130] Table 9 Comparison of neurotransmitter content in each group of samples before treatment

[0131]

[0132] Note: Compared with the Sham group, aP<0.05; compared with the sham group, bP<0.05; compared with the traditional group, cP<0.05;

[0133] (2) Comparison of serum neurotransmitter levels before and after visual acupotomy

[0134] Compared with those before treatment, the levels of 5-HT, PGF2α, BK, NPY and CGRP1 decreased 7 and 14 days after treatment, and the differences were statistically significant (P < 0.05). There were no statistically significant differences in the levels of SP and TK 7 days after treatment (P > 0.05), but the levels of SP and TK decreased 14 days after treatment, and the differences were statistically significant (P < 0.05). Compared with those 7 days after treatment, the levels of 5-HT, SP, PGF2α, TK, BK, NPY and CGRP1 14 days after treatment showed statistically significant differences (P < 0.05), as shown in Table 10 below.

[0135] Table 10 Comparison of serum neurotransmitter levels before and after visual acupotomy treatment

[0136] neurotransmitters Before treatment 7 days after treatment 14 days after treatment F-number P-value 5-HT (ng / mL) 932.98±143.44 <![CDATA[535.21±65.70 * ]]> <![CDATA[325.74±33.19 *# ]]> 88.033 0.000 SP (pg / mL) 11097.33±388.06 9407.58±1096.66 <![CDATA[4919.72±540.48 *# ]]> 89.962 0.000 PGF2α (pg / mL) 38.17±8.38 <![CDATA[24.25±1.95 * ]]> <![CDATA[19.86±1.04 *# ]]> 25.479 0.003 TK (ng / mL) 1.47±0.19 1.24±0.16 <![CDATA[0.88±0.15 *# ]]> 35.314 0.000 BK (pg / mL) 12.57±1.85 <![CDATA[9.91±0.74 * ]]> <![CDATA[6.84±0.58 *# ]]> 57.081 0.000 NPY (ng / mL) 41.19±6.28 <![CDATA[28.83±2.15 * ]]> <![CDATA[21.08±1.03 *# ]]> 62.170 0.000 CGRP1 (pg / mL) 153.16±12.86 <![CDATA[127.92±12.48 * ]]> <![CDATA[103.33±5.84 *# ]]> 47.562 0.000

[0137] Note: Compared with before treatment, *P < 0.05; compared with 7 days after treatment, #P < 0.05;

[0138] (3) Comparison of serum neurotransmitter levels before and after traditional acupuncture treatment

[0139] Compared with those before treatment, the levels of 5-HT, SP, PGF2α, TK, BK, NPY and CGRP1 decreased at 7 and 14 days after treatment, and the differences were statistically significant (P < 0.05). Compared with those at 7 days after treatment, the levels of 5-HT, SP, PGF2α, TK, BK, NPY and CGRP1 at 14 days after treatment were statistically significant (P < 0.05), as shown in Table 11 below.

[0140] Table 11 Comparison of serum neurotransmitter levels before and after traditional acupuncture treatment

[0141] neurotransmitters Before treatment 7 days after treatment 14 days after treatment F-number P-value 5-HT (ng / mL) 1034.53±127.62 665.98±.38* 402.82±35.91*# 85.614 0.000 SP (pg / mL) 10931.37±915.65 8533.59±371.48* 6566.19±966.77*# 42.456 0.000 PGF2α (pg / mL) 38.45±7.01 25.99±1.65* 22.04±0.97*# 30.068 0.002 TK (ng / mL) 1.51±0.15 1.37±0.13* 1.08±0.12*# 83.477 0.000 BK (pg / mL) 12.73±1.19 10.32±0.84* 8.23±0.85*# 140.583 0.000 NPY (ng / mL) 38.64±7.26 28.07±1.81 23.51±1.44*# 21.756 0.005 CGRP1 (pg / mL) 165.10±11.87 128.11±13.82* 108.01±8.37*# 75.771 0.000

[0142] Note: Compared with before treatment, *P < 0.05; compared with 7 days after treatment, #P < 0.05;

[0143] (4) Comparison of neurotransmitter levels between the visual acupotomy group and the traditional acupotomy group after treatment

[0144] After treatment, the two groups showed that the levels of 5-HT, SP, PGF2α, TK, BK, NPY and CGRP1 in the visual acupotomy group were lower than those in the traditional group. Except for CGRP1, the differences in other neurotransmitters were statistically significant (P < 0.05). The results are shown in Table 12 below.

[0145] Table 12 Comparison of neurotransmitter content between visualization group and traditional group after treatment

[0146]

[0147] 4. Exploring the molecular mechanism of visual acupotomy intervention for pain trigger points in the treatment of “stasis pain” in ONFH model animals

[0148] 1. High-throughput screening of trigger point intervention for the treatment of osteonecrosis of the femoral head using visual acupotomy revealed a complex metabolic crisis interaction network in ONFH: The experiment used gene chips to detect differential efficacy genes with a trigger point difference of 1.5 and found that there were 685 intersection genes that were downregulated in the model group compared with the sham surgery group and upregulated in ONFH compared with the model group, and 142 intersection genes that were upregulated in the model group compared with the sham surgery group and downregulated in ONFH compared with the model group. Gene classification revealed that the mechanism of acupotomy efficacy was involved in pathological processes such as energy metabolism, muscle metabolism, microcirculation, herniation transmission and pain, inflammation and oxidative stress, suggesting that osteonecrosis of the femoral head has a metabolic crisis in the above pathological links, rather than a single pathological link, and that the various networks are not isolated;

[0149] (2) Low-throughput screening and network construction of neurotransmitters revealed that ONFH has a complex neurotransmitter metabolic network: the experiment selected a low-throughput screening method to detect choline, amino acids, monoamines, peptides, prostaglandins, histamine, nucleotides, endothelial relaxing factor, etc., and found that the metabolic disorders of the above neurotransmitters were detected. Visual acupuncture can correct the above disorders. The gene-metabolite or string-based protein interaction networks of peptide neurotransmitters were constructed through the metascape plug-in of cytoscape, and it was found that ONFH has a complex neurotransmitter-gene metabolic interaction network.

[0150] (3) The intersection of visual acupuncture knife differential efficacy genes and neurotransmitter metabolism network revealed ACHE, PNMT, and MAOB, which may be the core targets of visual acupuncture knife treatment for ONFH "pain and stasis": the intersection of up-regulated gene 685 and non-peptide neurotransmitter network revealed 10 intersection genes, including FPGS, POLR2E, POLR2I, ACHE, OPLAH, PNMT, ATP1A4, ATP6V0C, ALDH4A1, and NME3; the intersection of up-regulated gene 685 and peptide neurotransmitter network revealed one intersection gene, RAMP1; the intersection of down-regulated gene 142 and non-peptide neurotransmitter network revealed three genes, including ADCY5, MAOB, and GLYAT, and the down-regulated gene 142 had no intersection with the peptide neurotransmitter network. Further analysis revealed that downregulation of ACHE and PNMT is consistent with the pathology of ONFH pain trigger points. Downregulation of both can increase the acute state of trigger points and lead to a series of energy, microcirculation, inflammation, oxidative stress, pain, and synaptic transmission disorders. MAOB plays an important role in the metabolism of neurotransmitters such as neuroactive amines and vasoactive amines in peripheral tissues, suggesting that visual acupuncture can regulate pain by reducing MAOB at pain trigger points and regulating neurotransmitter metabolism disorders.

[0151] (4) Verification of ACHE, PNMT, and MAOB, the core targets of visual acupuncture for ONFH "pain and stasis": The reduction of ACHE and PNMT can lead to excessive accumulation of acetylcholine and catecholamines, maintaining the continuous discharge of neuromuscular junctions, muscle spasms and circulatory disorders; and the increase of MAOB in the model group suggests its role in correcting the imbalance of neurotransmitters at the pain trigger point; the experiment further used ACHE, PNMT, and MAOB agonists to find that there was no difference in the mechanical pain sensitivity threshold of the pain trigger point between the acupuncture intervention group and the agonist group, and there was no significant difference in the related inflammatory factors and neurotransmitters compared with the visual acupuncture group, suggesting that ACHE, PNMT, and MAOB are the core targets of visual acupuncture for ONFH "pain and stasis".

[0152] 5. Clinical validation of visual acupotomy in the treatment of ONFH "pain and stasis"

[0153] In order to verify the existence of "neurotransmitter-muscle fiber metabolic disorder-circulatory disorder" interaction in the pain trigger points of patients with femoral head necrosis, and to clarify the effect of visualized acupuncture knife on improving the "stasis and pain" of femoral head necrosis, the present invention further added clinical verification of visualized acupuncture knife treatment for ONFH "stasis and pain" in addition to completing the established research content. 26 patients with femoral head necrosis who received acupuncture knife treatment from August 2020 to November 2021 were collected. There were 19 males and 7 females, aged 21 to 64 years, with a median of 44 years. According to the Association Research Circulation Osseous (ARCO) staging standards for ONFH, there were 2 cases in ARCO stage II, 14 cases in ARCO stage III, and 10 cases in ARCO stage IV. The visual analogue scale (VAS) score of hip pain, hip joint range of motion (flexion, abduction, internal rotation, external rotation), modified Ashworth score, cord index score, serum levels of pain-related inflammatory factors, vasomotor factors, and neurotransmitters were recorded before surgery and 1 day and 7 days after surgery to verify the therapeutic effect of acupotomy and explore the mechanism of action of visual acupotomy therapy in improving the "pain and stasis" of femoral head necrosis.

[0154] 1. The effect of visual acupuncture therapy in relieving the "pain and stasis" of femoral head necrosis

[0155] The analysis results of VAS scores before and after visual acupuncture treatment showed that the VAS scores at two observation time points, 1 day and 7 days after visual acupuncture treatment, were lower than those before treatment (p<0.01), as shown in Table 13 below.

[0156] Table 13 Comparison of VAS scores before and after visual acupotomy treatment ( point)

[0157] Observation time point VAS (points) t-value P-value Before treatment 5.27±1.62 1 day after treatment 4.56±1.20 3.364 0.002 7 days after treatment 3.08±1.09 11.060 0.000

[0158] 2. The effect of visual acupotomy on improving joint function of femoral head necrosis

[0159] The analysis results of the total range of motion of the hip joint before and after visual acupuncture knife treatment showed that the total range of motion of the hip joint at the two observation time points of 1 day and 7 days after visual acupuncture knife treatment was greater than that before treatment (p<0.01), see Table 14 below.

[0160] Table 14 Comparison of total range of motion of the hip joint before and after visual acupotomy treatment ( Spend)

[0161] Observation time point Total range of motion of the hip joint (degrees) t-value P-value Before treatment 163.28±31.12 1 day after treatment 188.97±30.22 -9.383 0.000 7 days after treatment 217.07±30.87 -13.479 0.000

[0162] 3. The effect of visual acupuncture therapy on reducing the tension of local muscle tension in the hip joint

[0163] The analysis results of the induration cord index and muscle tension before and after visual acupotomy treatment showed that the induration cord index and muscle tension at two observation time points, 1 day and 7 days after visual acupotomy treatment, were lower than those before treatment (p<0.01). The results are shown in Tables 15 and 16 below.

[0164] Table 15 Comparison of induration cord index before and after visual acupotomy treatment ( point)

[0165] Observation time point Induration cord index (points) t-value P-value Before treatment 2.77±0.43 1 day after treatment 1.81±0.40 25.000 0.000 7 days after treatment 1.27±0.60 13.117 0.000

[0166] Table 16 Comparison of Ashworth scores before and after visual acupotomy treatment ( point)

[0167] Observation time point Ashworth scale (points) t-value P-value Before treatment 4.46±0.65 1 day after treatment 3.31±0.68 15.990 0.000 7 days after treatment 3.00±0.69 14.659 0.000

[0168] 4. Comparison of inflammatory factors before and after visual acupotomy treatment

[0169] The results of Capsaicin before and after treatment showed that the Capsaicin content increased one day after treatment compared with that before treatment (P>0.05). Visual acupuncture is an invasive treatment that cuts local soft tissue and causes a transient inflammatory reaction. The Capsaicin content was lower than that before treatment 7 days after visual acupuncture treatment (P<0.05). The results of PGF2α, TNF-α and IL-1β before and after treatment showed that the PGF2α, TNF-α and IL-1β contents were lower than that before treatment at two observation time points: one day after visual acupuncture treatment and 7 days after visual acupuncture treatment. The difference between 7 days after treatment and before treatment was statistically significant (P<0.05). This shows that visual acupuncture therapy can relieve the pathological tension of trigger points and achieve the therapeutic effect of reducing local inflammatory reactions. The pain-relieving effect of visual acupuncture therapy is closely related to its reduction of serum inflammatory factor levels and inhibition of local inflammatory response process. The results are shown in Table 17 below.

[0170] Table 17 Comparison of inflammatory factor levels before and after visual acupotomy treatment

[0171]

[0172]

[0173] Note: a means P<0.05, b means P>0.05

[0174] 5. Comparison of vasomotor factors before and after visual acupotomy treatment

[0175] The results of NO before and after treatment showed that the NO content at two observation time points, 1 day and 7 days after visual acupotomy treatment, was higher than that before treatment (p<0.05). Visual acupotomy can cut local adhesions, scars and contracture tissues, which can reduce the tension of local tissues in the hip joint and improve local blood circulation. Vascular dilation may be closely related to the continuous increase in NO content after visual acupotomy and the dilation of arteriolar blood vessels. The results of ET-1, HIS and NA before and after treatment showed that ET-1, HIS and NA were lower than those before treatment 7 days after visual acupotomy treatment (p<0.05). There was a statistically significant difference in HIS 1 day after treatment compared with before treatment (p<0.05). There was no statistically significant difference in ET-1 and NA 1 day after treatment compared with before treatment (P>0.05). ET-1, HIS, and NA are important mediators of inflammatory pain. The levels of these vasomotor factors in serum were reduced to varying degrees after visual acupotomy treatment, and the reduction effect was most obvious 7 days after surgery. The above results indicate that the effect of visual acupotomy in reducing local inflammatory response and relieving pain is closely related to the regulation of vasomotor factors. The results are shown in Table 18 below.

[0176] Table 18 Comparison of inflammatory factor levels before and after visual acupotomy treatment

[0177] Group NO ET-1 NA HIS Before treatment 4.09±1.08 2.58±0.79 0.47±0.10 9.48±2.41 1 day after treatment 5.11±1.13a 2.30±0.59b 0.46±0.95b 8.85±2.01a 7 days after treatment 5.38±1.08a 1.79±0.54a 0.40±0.94a 7.39±1.97a

[0178] Note: a means P<0.05, b means P>0.05

[0179] 6. Comparison of neurotransmitters before and after visual acupuncture treatment

[0180] The results of CGRP1, SP, BK, and NPY before and after treatment showed that the levels of CGRP1, SP, BK, and NPY were all lower 7 days after visual acupotomy treatment than before treatment (p<0.05). The difference in SP was statistically significant 1 day after treatment (p<0.05). There was no statistically significant difference in CGRP1, BK, and NPY 1 day after treatment (P>0.05). The above neurotransmitters are important inflammatory mediators or pain factors. Their serum levels were significantly reduced after visual acupotomy treatment, indicating that the pain-relieving effect of visual acupotomy is closely related to its reduction of serum levels of neurotransmitters that inhibit pain and inflammation. The results are shown in Table 19 below.

[0181] Table 19 Comparison of neurotransmitter levels before and after visual acupotomy treatment

[0182] Group BK NPY CGRP1 SP Before treatment 1.74±0.62 167.00±56.56 36.71±13.37 280.10±51.74 1 day after treatment 1.65±0.48b 152.11±46.47b 33.99±15.63b 249.36±34.46a 7 days after treatment 1.41±0.40a 125.06±35.52a 29.09±10.85a 228.65±31.78a

[0183] Note: a means P<0.05, b means P>0.05

[0184] In summary, the present invention found that acupuncture knife therapy can achieve the purpose of treating ONFH pain by regulating the expression of vasomotor factors and promoting the repair of femoral head blood supply. The present invention found that ONFH has a complex abnormal correlation network of energy metabolism, muscle metabolism, microcirculation, pain synaptic transmission, inflammation and oxidative stress by combining high-throughput screening of differential genes with low-throughput screening of neurotransmitter detection and gene-metabolite network construction and analysis. The intersection analysis of the metabolic network and differential genes after low-throughput neurotransmitter screening and experimental verification found that ACHE, PNMT, and MAOB are the core targets of visual acupuncture knife therapy for ONFH "stasis pain", indicating that correcting the metabolic imbalance of pain trigger points acetylcholine, norepinephrine, vasoactive amines, and neuroactive amines and then improving microcirculation is the main mechanism of action of visual acupuncture knife therapy for ONFH "stasis pain".

[0185] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to the embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the claims and their equivalents.

Claims

1. A method for studying the mechanism of action of visual acupotomy in treating femoral head necrosis and pain, characterized in that: The following steps are involved: S1. Establish an animal model of femoral head necrosis; S2. Based on the femoral head necrosis animal model established in step S1, a femoral head necrosis pain trigger point model is established; S3, locating and marking the femoral head necrosis pain trigger points on the femoral head necrosis pain trigger point model obtained in step S2; S4. The femoral head necrosis pain trigger point models marked in step S3 are divided into a model control group, a traditional acupotomy group, and a visual acupotomy group. The expression levels of inflammatory factors, the expression levels of vasomotor factors, and the content of neurotransmitters in the serum of the animals in each group are tested before and after the intervention with traditional acupotomy and visual acupotomy, respectively, to obtain the test results. S5. Using gene chips, high-throughput screening is performed on the model control group and the visual acupotomy group obtained in step S4 to obtain genes with differential therapeutic effects; S6. Performing low-throughput neurotransmitter screening and network construction on the model control group and the visual acupotomy group obtained in step S4 to obtain a neurotransmitter-gene metabolism interaction network; S7. Combining the detection results of step S4, the differential efficacy genes of step S5 and the neurotransmitter-gene metabolism interaction network of step S6, the potential core targets of visual acupotomy in treating pain and bruise of femoral head necrosis are obtained, that is, the mechanism of action of visual acupotomy in treating pain and bruise of femoral head necrosis is clarified.

2. The research method of the mechanism of action of the visualized acupotomy in treating femoral head necrosis and pain according to claim 1 is characterized in that: Step S1 includes the following steps: S11. After anesthetizing the beagle dog, one forelimb was depilated and routinely disinfected. A Kirschner wire was used to fix the carpometacarpal joint of the dog through an anterior incision. The end of the Kirschner wire was buried subcutaneously and sutured. The dog recovered for 2 weeks to obtain a three-legged beagle model. S12. Depilate one side of the buttocks of the three-legged beagle model obtained in step S1. After routine disinfection, a 3.5 mm diameter bone tunnel is drilled from the greater trochanter of the femur to the femoral head, 2 mm below the surface of the articular cartilage. After fluoroscopic confirmation of a satisfactory position, 0.5 MPa liquid nitrogen is perfused into the femoral head and frozen for 2 minutes, then rewarmed for 2 minutes. The freezing and rewarming procedure is repeated 4 times for a total of 10 minutes. The wound is closed and recovered for 14 days to obtain an animal model of femoral head necrosis.

3. The research method of the mechanism of action of the visualized acupotomy in treating femoral head necrosis and pain according to claim 1 is characterized in that: Step S2 includes the following steps: The animal model of femoral head necrosis established in step S1 was subjected to eccentric treadmill exercise once a week with a treadmill inclination angle of 16° and a speed gradually increased to 12 km / h for 60 minutes. The animal was fed normally for the remaining 6 days without eccentric exercise for 8 weeks, followed by a recovery week. The animal was fed normally during the recovery period without any experimental intervention to obtain a femoral head necrosis pain trigger point model.

4. The research method of the mechanism of action of visual acupotomy in treating femoral head necrosis and pain according to claim 1, characterized in that: Step S3 includes the following steps: The femoral head necrosis pain trigger point model obtained in step S2 is anesthetized, and the femoral head necrosis pain trigger point model is placed in a supine position with the limbs fixed. The femoral intertrochanteric crest, rectus femoris, sartorius muscle, gluteus medius, and adductor muscle origin and insertion points are touched in turn. If an enlarged nodule is touched, it is determined to be the femoral head necrosis pain trigger point and marked.

5. The research method of the mechanism of action of visual acupotomy in treating femoral head necrosis and pain according to claim 1, characterized in that: In step S6, the neurotransmitter-gene metabolism interaction network was constructed by constructing the gene-metabolite interaction network using the metascape plug-in of cytoscape, and the protein interaction network of peptide neurotransmitters was constructed using string.

6. The research method of the mechanism of action of visual acupotomy in treating femoral head necrosis and pain according to claim 1, characterized in that: In step S7, the potential core targets are ACHE, PNMT and MAOB genes.

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