Study On The Effect Of Stretching To Fascial Remodeling And Different Transcription Level Of Remodeling-Related Gene

Background:Traditional Chinese medicine for thousands of Chinese culture as a crystallization of Chinese civilization in the process of upper and lower transmission plays an important role in Chinese science and technology to prevent and cure diseases,especially of the time developed an indispensable process of preventive health care,are confirmed in practice scholars of the ancient Chinese medicine experience lofty ideals,for which won the admiration of the public.In recent years, with the development of technology,cultural exchanges and people to accelerate the treatment of Green's request,in theory under the guidance of traditional Chinese medicine treatments not only in the Chinese residential area but also in Europe and the United States developed regions such as the United Kingdom,Germany,France, Belgium,the Netherlands,the United States,Canada extensively as a supplement or alternative treatment which is carried out.In recent years,with the development of technology,cultural exchanges and people to accelerate the treatment of Green's request,treatments under the guidance of traditional Chinese medicine are carried out not only in the Chinese residential area but also in Europe and the United States developed regions such as the United Kingdom,Germany,France,Belgium,the Netherlands,the United States,Canada extensively as a supplement or alternative treatment.As the one of core of traditional Chinese medicine,the meridian theory,or the needle,moxibustion practice under the guidance of the meridian theory attract vast numbers of scholars to study it.Recalling the history of acupuncture meridians study,the has two climax period in abroad from fifties to sixties of twentieth century and from the end of the twentieth century to now.Japan(Ryukyu countries) and Korea(Korea country) exchang culture with ancient china closely,but the "Golden Wind Han," Case stop the enthusiasm and led to the climax to End.Because of economy,technology,talented scholars,the United States far away in the western hemisphere had paied much attention to Acupuncture and Meridian Research and now they have a number of research papers published in SCI and had raised several hypothesis.As China,we have the duty to Clarify the mechanism of acupuncture and meridians.Therefore,our country has invested substantial funds and a large number of scholars to study it,and has established "eight - five" and "nine - five" meridian climb research projects.Clinical,laboratory researchs confirmed the objective existence of meridians,and further found that Meridian has a cross-section of conductive body through by a two-way transmission,and low resistance,high conductivity,high temperature luminescence,and calcium ion concentration of characteristics of the population.Summed up decades researching,the had difficulies to unify the various theories.The biggest rift exeisted between "functional" and "structure","overall" and "partial".Since the HM Langevin proposed the standpoint of loose connective tissue in the human body constitutes a "flaky network support" and "signal network",the researchers come to realize the importance of structure in our bodies.It is worth mentioning that the Professor Yuan Lin Research Group think that fascia in the human body from the shallow into the deep structure has five kinds of components:1,dermal dense connective tissue;2,subcutaneous loose connective tissue;3,muscle surface of loose connective tissue;4,muscle and muscle clearance interval connective tissue;5,door internal organs,connective tissue envelope and internal partitions.Group by studying the human body 14 GB 361 meridian acupoints needling sites and methods and found that parts of the body acupuncture points are located at different levels of the fascia,which is located in muscle connective tissue gap spacing and muscle up,followed by dense dermal connective tissue layer and the subcutaneous loose connective tissue layer,a small number of loose connective tissue at the muscle surface(deep fascia) and the internal organs door,envelope and internal partitions of connective tissue.By combination the above-mentioned studies and the developmental biology,Professor Yuan Lin research group pointed out:in multicellular organisms,mesenchyme is derived from jelly-like extracellular fluid (ECF) and further differentiates into many cell types,tissues,even organs.In higher creatures like human beings,mesenchyme gives rise to all the fascial tissue throughout the body.The fasciae form support the a framework of connective tissues to growth of cells of all except for framework the central nerve system,can be regarded as one functional organs and therefore this of the essential elements of the internal environment for all kinds of living cells.Functionally,the connective tissue framework not only provides a soft support of the body but also maintains the systemic balance by regulating the neural reflexes, neuroendocrine activity,neuroimmunity,and by repairing the cell and tissue injuries. Therefore,they put forward a hypothesis that an auto-surveillance system may exist in the human body,which differs essentially from the nine established functional systems.The auto-surveillance system has two main functions. One is to regulate the functional state(metabolic activity and excitation),such as neural reflex regulation and neuroendocrine regulation.The other is to regulate the state of life activity(cell division,growth and apoptosis),including cell division and growth regulation through the sympathetic nerves and cell apoptosis regulation through lymphocyte-mediated immunity.In human,the auto-surveillance system is composed of the connective tissue network which monitors and regulates living and functional conditions of tissue cells.The group's study by using ultrasound,CT,MRI and other medical imaging tools consider:In the course of acupuncture,by interpolation or twist,you can drive or affect a large scope of the connective tissue;When the acupuncture needle generally arrive outside the sarcolemma,a sense "de qi" occurs,when the needle arrival the muscles,the sense "de qi"weakened,if the needle stays on outer sarcolemma to increase the excitement,this sense will enhance;By using CT images,we have built strip structure wich run along the meridian.But goup also acknowledged the existence of inconsistent line structure.Contrast to the record in Chinese medicine books:group think that:oks,records related to acupuncture points:Systemic connective tissue scaffold may be the material basis of the meridian;the acupoint is the site which can generate strong biological information in the connective tissue scaffold;the diference between acupoint and no-acupoint is only the the amount of information and not the t qualitative difference.From a broad perspective,all parts of the body are acupuncture points,because connective tissue exist almost all the body's tissue gap.Because in early studies,group pay more attention to looking for imaging evidence for meridian,so they know less about the cells' and fiber' reaction under stretch,and the different level of gene transcription.This is precisely the focus of this study.In this study,immunocytochemistry and molecular biology methods were used to discussed the reaction of fiber or cells and the level of related genes in loose connective tissue under stretch.Through this research,wo want to explore the material basis of the meridian,and provide theoretical support for the faciaology.Objective:By studying on the effect of stretching to fascia remodeling and different gene transcription level of remodeling-related gene,group want to provide evidence to the relevance of meridians and connective tissue;to provide theoretical support for faciaology;and to explore the therapy mechanism for supplement therapy.1.By studying the effect of stretching to connective tissue fiber morphology,group want to explain the role of collagen or elastic fibers of connective tissue in the process of stretching.2.By studying the effect of stretching to connective tissue cells' morphology,group want to discussion the biological behavior of fascia connective tissue cells under the tensile stress.3.By studying the effect of stretching to connective tissue remodeling-related gene transcription level,group want to discussion the possible biological significance after stretching.Methods: 1.After stretching stimulation,subcutaneous loose connective tissue was cut,and then they were placed under microscope in order to observe the direction of fibert.2.tissue After stretching stimulation,subcutaneous loose connective was cut and stained by phalloidin and Pi,then they were placed under fluorescence microscope in order to observe the changes of cells' morphology under tensile.3.Mechanical tensile stimulate fascia connective tissue compartment;amplify gene fragments by RT-PCR,to observe the following genes' expression in the fascia connective tissue:CTGF,TGF-β1,Procollagen-1,Procollagen-3,MMP-2,MMP-3,TIMP-1,TIMP-2.Results:1,After Stretching,connective tissue fibers are arranged in the direction of force.2,After Stretching,connective tissue cell transformed into a long spindle shape, cells' long axis are arranged in the direction of force.3,After Stretching,the transcription level of CTGF,TGF-β1,TIMP-1 was decreased(Fvalue:4.774,6.151,15.877P＜0.05,Pvalue:0.023,0.011,0.000), whereas it have no influnce on Procollagen-1,Procollagen-3,MMP-2,MMP-3,TIMP-2(Fvalue:0.369,0.252,2.466,1.608 1.440,Pvalue:0.695,0.779,0.115,0.219,0.265).Conclusion:1.The group imposed with tensile stress by acupuncture,when needles were twisted, collagen fibers or elastic fibers winding around the filiform needle by electrostatic interaction between the filiform needle and collagen fibers or elastic fibers;stretch can cause deformation on connective tissue,but the control group maintain its original arrangment.2.By stretch,morphological deformation occur in collagen fiber or elastic fiber changed,so mechanical signals can transmitted into the cells through focal adhesion complex structure,causing cytoskeletal remodeling,cell transform into spindle shape.3.Under stretch,regulation the level of gene transcription of CTGF/ TGF-β1/TIMP-1 may be one of the mechanisms of complementary therapy.In conclusion,the group used acupuncture to impose sress,through electrostatic coupling between needle and connective,the fiber in connective tissue (primarily collagen fibers) winding around the needle,and then result in deformation of connective tissue,a direct result of the connective tissue deformation is driven the cells which contact with collagen fiber or elastic fiber by adhesion complexes.Inside the cells,through cytoskeletal remodeling,the stress signal transduce to the cell's nucleus,causing a series of changes,including the regulation of cell activity and the regulation of gene.Fibroblasts are the main cells in connective tissue,which involved in the process of repair,at the same time,fibroblasts can also synthesize a variety of cytokines,including the formation or degradation of matrix factor;Matrix formation or degradation directly affect the basement membrane permeability,which closely relate to many diseases,for example the cancer.Mesenchymal stem cells as a pluripotent stem cell,have the ablity to multi-directional differentiation,so the differentiated cells can replace the dysfunctional cells,and involved in injury repair and functional reconstruction;after the stretch signals transmission into cell nucleus, it causes a series of gene transcription change,such as CTGF / TGF-β1/TIMP-1, these genes have confirmed plays an important role in body reconstruction after injure, and regulate the downstream gene.So the deformation of connective tissue under stretch maybe one mechanism of complementary therapy.It regulation the cells' activity and the level of gene transcription,it maybe the mechanism of complementary therapy.