| BackgroundTraditional acupuncture and moxibustion defines acupoints as a special part of the body surface of the human body,which is the path of the disease and the site of acupuncture and moxibustion.Although specific parts are mentioned here,the specific structure is not described in detail.Modern research believes that acupoints are a complex of a series of known structures,such as nerves,blood vessels,etc.,which are spatially arranged and three-dimensionally constructed according to certain rules.Acupoints are the initial response sites for stimulation of the acupuncture and moxibustion,and their effects are inseparable from the structure and function of the skin.The skin is composed of epidermis,dermis and subcuits.They contain various cells with different physiological functions in different layers,such as keratinocytes,melanocyte,Langerhans cells(LCs)and Merkel cells.Skinimmune system(SIS)can be divided into two parts:cells and body fluids.Cell components include keratinocytes,LCs,dendritic cells,macrophages,T cells,neutrophils,mast cells and endothelial cells.The humoral components include peptide,complement,secretory immunoglobulin and cytokines.The exploration of acupoint structure has experienced several stages from macroscopic to microscopic recognition.In the past,researchers thought that most of the acupoints in the human body were located where the nerve endings were concentrated or there were thicker nerve fibers,but there was no definite conclusion about the specific structure.So far,we are still lack of in-depth and systematic research on the relevant organizational structure of acupoints.In the past,by visual observation of the surface and the partial dissection of the macro to the micro analysis,or with the help of all kinds of microscope,tissue section on microscopic image recording and analysis,we observed the acupoints are composed of multiple ingredients known organization structure,They are carriers that can be directly contacted or acted upon during acupuncture and moxibustion and thereby exert biological effects into full play.On the whole,in recent decades,researchers have focused on the study of acupoint structure mostly on the surface location of acupoints and its general organizational structure,but lack of in-depth study on the classification of cells and the chemical components they contain.Since we are not able to confirm the existence of the specific structure corresponding to the acupoints,it is the natural choice for the basic research on the biology of acupoints to study the cellular classification and chemical characteristics related to the acupoints with the known organizational structure.At present,with the development and progress of histological research techniques,more effective methods are provided for us to deeply study the microstructure of the acupoints at the histological level,and the conditions are created for us to better explore the relevant organizational structure of the acupointsThe part of the skin that exerts biological effects after acupuncture and moxibustion stimulation is closely related to its microcirculatory structure,nerve endings,and cellular bioactive components.Although the selection of acupoints and the way of stimulation play a vital role in the curative effect of acupuncture and moxibustion,a systematic and in-depth understanding of the biological basis and microstructure of acupoints is helpful to better explain the mechanism of acupuncture effect.What we need to pay attention to is that due to the difference in the location of the acupoints,there are also many differences in the thickness of the skin,the number of muscles and the density of the nerves.Therefore,in order to understand the biological basis of acupoints more deeply,we should focus on the histochemical research related to acupoint structure.After receiving the stimulation,the bioactive substances in the acupoints will change accordingly.It is this change that initiates the regulation of the body's homeostasis and the occurrence of a series of corresponding cascades,thus achieving the goal of curing the disease.This indicates that acupuncture and moxibustion have effects of regulating and maintaining human function homeostasis.Acupuncture and moxibustion stimulation can make the skin at the acupoint feel a sense of needle which is so-called "Dei qi" and "passing through the senses".The excitement can be transmitted to the central body through various structures in the acupoint.The excitatory impulses of the sympathetic nerve endings around the vessel wall play a key role in the generation of the sensory transmission and the "acupoint-viscera-related" regulation effect.The density of local nerve fibers,the distribution of blood vessels and lymphatic vessels,the activation of various immune cell functions and the production of related chemicals are the biological basis of acupuncture effect.As a local immune microenvironment,blood vessels and lymphatic vessels are the premise of acupoint immunogenesis:when the body is damaged,some chemical active substances will appear in the lymphatic fluid,such as inflammation and vascular reaction mediators,which may play a key role in acupuncture effect.At the same time,acupuncture and moxibustion generate mechanical and chemical stimulation on local tissues of acupoints,which may transmit acupuncture and moxibustion signals via the upline system to the central nervous system with the aid of peripheral receptors and free nerve fibers,and then regulate the functions and activities of the body and organs through the integration and reflection of various central organs.As an important part of the skin neuro-immune-endocrine network,humoral regulation and cellular immunity involved in lymphatic vessels and microvessels play an important role in explaining the mechanism of acupuncture.Therefore,it is suggested that the correlation between nerve fiber,blood vessel,lymphatics and immune cells in local tissues of the acupoints should be studied as a key point to explain the meridian effect of acupuncture and moxibustion at the cellular level.Communication between the brain and the skin is mediated by hormones in the endocrine system,each of which plays its own regulatory role.Mast cells are not only target cells of various hormones,but also release many inflammatory mediators,which plays a key role in mediating the neural-immune-endocrine network.The brain communicates with the skin through the hypothalamic-pituitary-adrenal(central HAP axis)and the sympathetic adrenal medulla(SAM axis).At the same time,different parts of the skin can also secrete similar hormones,forming a microenvironment of local regulation of the skin(HAP axis).Mast cells are ubiquitous in the human body,mainly distributed around the blood vessels,especially in the contact parts with the blood vessels and nerves.They can make specific responses to the neuropeptides produced by nearby neurons,activate degranulation and induce the production of cytokines and chemokines.After receiving acupuncture signals,mast cells release inflammatory mediators through degranulation to convert mechanical stimulation into biological signals,and then through the integration and transmission of neuro-endocrine-immune network,the feedback regulation can be applied to target organs,thus achieving the effect of curing disease.The joint parts of acupuncture and moxibustion are acupoints,but the two methods of stimulation are different.Moxibustion mainly acts on the human body through the thermal stimulation of the burning of moxa,while acupuncture is mainly mechanical stimulation of the needle with the application of manipulation.Acupuncture and moxibustion can not only increase the number of local mast cells in the acupoint area,cause the effect of meridian aggregation,but also stimulate the degranulation,release histamine,5-hydroxytryptamine,heparin and so on.These mediators can not only increase vascular permeability,but also cause local edema and mild spontaneous inflammation.On the one hand,it may promote mast cells to gather at acupoints in the lesion by inducing adhesion molecules and chemokines.On the other hand,it activates the immune system and broadens the effects of local acupuncture throughout the body.Objective1.By studying the mechanical pain threshold of the skin at different acupoints and non-acupoints,it was revealed whether there were sensory differences between different acupoints and non-acupoints.2.By observing the interaction between nerve fiber vessels and immune cells in the relevant local tissues of the acupoints,the relationship between the structure and function of the relevant histochemistry of the acupoints was revealed.3.By studying the effects of acupuncture and moxibustion on the activation state of mast cells in the acupoint area,the differences in the effects of moxibustion and acupuncture on the body were revealed.Materials and methodsPain threshold detection:According to the literature,the location of acupoint area is the skin and muscle tissue of 1 cm x 1 cm centered on acupoint,while the location of non-acupoint area is the skin and muscle tissue of 1 cm × 1 cm at 1.5 cm on the left or right side of acupoint.Determining the non-acupoint control of this experiment by the conversion of the corresponding ratio of human and rat.Selecting representative acupoints from the head,trunk and limbs of the rat:Non-acupoint 1(midpoint of the connection between the DU20 and BL2 of the rat),GB14(located above the center of the rat's eye,in the depression above the bone),Non-acupoint 2(5 mm next to LI11 of the rat),SJ5(located above the wrist 3mm,between the ulna and the radius).GB30(located in the posterior border of the hind limb hip joint),LR03(located in the depression between the first and second metatarsal bones of the hind foot).After 30 minutes of rest,the rats were tested,and the stimulus needles of the hand measuring apparatus stimulated the skin of different acupoints and gradually increased the intensity of stimulation until the rats had a withdrawal reaction,and the intensity of stimulation was recorded.The interval between the two tests should be 10 min.Measure three times and record the data of each time.Comparative study of acupuncture and moxibustion:A total of 24 male Sprague Dawley rats(220±20 g)were randomly divided into Control,Acupuncture and Moxibustion groups(n=8 in each group).After anesthesia of rats,In the acupuncture group,0.25 mm×15 mm acupuncture needles were directly penetrated into the skin of LR03 for 1.5 mm.The needles were continuously rotated at a small range for 10 min at a uniform frequency(120 times/min).In the moxibustion group,incense sticks were used instead of moxa sticks to keep the local skin temperature of the moxibustion point at 41±0.2 ?,continuous moxibustion for 10 min.At the end of the intervention,the rats were sequentially filled with 0.9%sodium chloride solution and fixative containing 4%paraformaldehyde and 0.1 mol/L phosphate buffered solution(PB,pH 7.4).The skin tissue of the acupoints was taken out and made into tissue sections.Immunofluorescence histochemical staining was performed and observed by confocal microscopyResultsPain threshold test results:There was no significant difference in pain threshold between acupoints and non-acupoints(GB14 and non-acupoint 1,SJ5 and non-acupoint 2)(P>0.05);the pain thresholds of acupoints and non-acupoints in the head and extremities(GB14,non-acupoint 1,LR03)are significantly lower than those in the trunk and near torso(SJ5,non-acupoint 2,GB30),the difference is statistically significant(P<0.05).The pain threshold of GB30 was significantly higher than that of other acupoints and non-acupoints(P<0.01).The results of acupuncture and moxibustion comparison:After acupuncture and moxibustion intervention,the contents of NPY,TH and CGRP positive nerve fibers in the skin tissue of rats in LR03 area increased obviously,which indicated that the differential regulation of the two on the local tissue of the point area was reflected in the regulation of neuropeptide;Acupuncture and moxibustion could promote the activation of mast cells(P<0.05),but there was no significant difference between them(P>0.05).The degranulation rate of activated mast cells in acupuncture group was significantly higher than that in blank control group(P<0.05),but there was no obvious difference between moxibustion group and acupuncture group.Acupuncture and moxibustion could promote the expression of 5-HT and HA(P<0.05),but there was no significant difference between them(P>0.05).Conclusions1.The pain threshold of the distal acupoints of the head and limbs was significantly lower than that of the trunk;There was no significant difference in pain threshold between acupoints and non-acupoints.2.Both acupuncture and moxibustion stimulation can enhance the expression of NPY,TH and CGRP in the epidermal nerve fibers,thereby activating mast cells and generating a cascade effect by releasing 5-HT and HA.These neuroactive components may play a mediating role in amplifying and transmitting acupuncture signals and exert biological effects.3.Both acupuncture and moxibustion can activate mast cells in local tissues of the acupoint and promote degranulation,and acupuncture is better than moxibustion in degranulation of mast cells. |
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