Research About The Influence Of Specificity Of The Proteins Expression By Needling Taichong Acupoint

BackgroundPrimary hypertension is defined as a hypertensive vascular disease with ascending of arterial pressure. While the pathogeny and etiology are not clear, it is common regarded as a result of multiple factors. It increases the damage of cardiovascular systems and threats the health of mankind. Although antihypertensive therapy that modern medicine provides clearly reduces the risks of cardiovascular and renal disease. Insomnia of some patients and symptoms like dizziness and headaches are not improved. Meanwhile the side effects are still obvious. Clinical trials show that diuretics may increase the frequency of sexual dysfunction in men and women. Other kinds of antihypertensive drugs may cause changes in LDL and probably increase the risk for diabetes. The drug may also leads to peripheral edema. Moreover, considering primary hypertension a chronic disease, the long term implication may increase the economic burden on patients, their family and on the whole society, especially in rural areas and third-tier cities of China, where the public health insurance policy is not complete yet.Traditional Chinese Medicine (TCM) classics have no records of the name of "primary hypertension" it is classified as "dizziness, liver wind or headaches " by TCM physicians today according to the symptoms and signs of patients. As an essential part of TCM, acupuncture therapy performs effectively on treating primary hypertension.especially on mediating SBP in a short time. Researches also prove that symptoms like dizziness and headaches on patients improve better than common drugs. Acupuncture therapy also shows almost no side effects. This therapy is supposed to be a routine treatment for hypertension for its simplicity and high cost performance.However, the elusive mechanism limits the promotion of acupuncture in some degrees.It’s vital for us to study the unknown mechanism in recent days. Acupuncture based on traditional Chinese meridian theory. The theory considers human being as an organic entity. The various organs, tissues, and other parts are connected by the pathways named meridians. Recent investigations believe that the positive impact of acupuncture can be explained by multiple target effect which makes it more challenge to understand the mechanism of acupuncture and the prescriptions of acupoints. Meridian-cerebrum hypothesis thinks all of the meridians connected with brain. The cerebrum can change and integrate the acupuncture signals and then the signals will acts on certain targets to adjust functions and organs. Base on this hypothesis, we suppose that the CNS(central nerve system), PNS(peripheral nerve system) and humoral factor should have some connections, while the connection is not clear yet, this trail will focus on CNS.This trail belongs to Professor Lai Xinsheng, my PHD tutor’s National Natural Science research. Previous tests have already proved that acupuncturing on taichong point can apparently lower the BP of SHR M, compared with the model group, though it didn’t adjust BP to the normal level. But points like ququan, chongyang, usually believed to improve symptoms like headaches and dizziness did not work well. This also confirm the specificity of acupoints. Meawhile,it’s believed that oblongata has a close relation with BP, because there are multiple nerve nuclei and blood vessels through it. We are unclear about if acupuncture could effect oblongata to adjust BP and the protein channels of the target process. So we chose oblongata as the object of the research to find out the specificity of acupoints with pressure reduction function in CNS.ObjectsBased on the obvious results which shows the specificity of Taichong, Chongyang, and Ququan in mediating blood pressure, to study the difference of expression of the protein in SHR oblongata when acupuncturing the same points(Taichong, Chongyang and Ququan) that also supposed to have effect and difference on improving symptoms like dizziness and headaches. With mass spectrometry and recombinant protein technology, we tend to find out the CNS specificity of various acupoints when mediating BP and to provide some hints to the research of the singularity of acupoints.MethodsRandomly divided 55 SPF 9-week male SHR into Taichong team, Chongyang team, Ququan team, non-acupoint team and model team. Each team has 11 rats.89-week male wistar rats compose the normal team.Acupuncture the Taichong team, chongyang team Ququan team and non-acupoint team with Taichong point, Chongyang point, Ququan point and non-acupoint for 5 minutes. When performing the treatment, we fixed the rats’head with socks. All the operation was performed at the waking state of the rats. The acupuncture teams rats were needled in respective acupoints in right and left for 5 minutes, taking twisting tactics 80±10 times every minute with twist magnitude 180° ±30. All of the operation were performed once a day before 12 o’clock for a week. The non-acupoint team and model team was fixed with the same methods without operation.At the seventh day, we separated the oblongata tissues and grinded them and extract protein from the tissues. Being separated by 2D-PAGE, the pictures of the proteins were scanned and analyzed to find out the differences among the groups. Then the results were checked by MS and searching the MASCOT date base.Results(1)Analyzed by Imagemaste 2D 5.0, the 2D-PAGE pictures shows difference of oblongata expression protein point among the groups.① Compared with model group, there are 15 different protein points increased in Taichong group while 11 decreased.② Compared with model group, there are 34 different protein points increased in Chongyang group while 28 decreased.③ Compared with model group, there are 14 different protein points increased in Ququan group while 13 decreased.④ Compared with model group, there are 33 different protein points increased in non-acupoint group while 27 decreased.⑤ Compared with the normal group, there are 13 different protein points increased in model group while 28 decreased.(2) Identification of the different protein points:216 different protein points were discovered among this experiment.Being limited by the funding, only the obvious protein points were identified and the replicated one rejected. 37 protein points remained here:① Compared with model group, there are 6 proteins increased in Taichong group: Protein LOC100911959:specific generalization, effect the development and growth of cell circle and transduction of the signals. Transitional endoplasmic reticulum ATPase:connected with the growth, apoptosis and repairmen of DNA process,high expression in some tumor cells and is connected with the periodization, classification and prognosis. MICOS complex subunit Mic60(fragment):connected with the control of the morphology of mitochondrial cristae and process of transformation of protein. Dihydropyrimidinase-related protein 3:connected with extension of neurite and formation of neuronal polarization. Part of dynamic regulation of the microtubules. Cathepsin B:part of the process that the lysosomes degrading the proteins and connected with the antigen processing and activation of hormone. Peroxiredoxin-2:the main cleaner of hydrogen peroxide in RBC, protect RBC and the cell membrane of it. 〤ompared with model group, there are 4 proteins increased in Chongyang group. AP-2 complex subunit beta:apolipoprotein subumits, component of plasma lipoproteins. Triosephosphate isomerase:Important part in the process of glycolysis isomerase, influence the energy generated effectively;lack of it can lead to immune disorder system disease. Endoplasmin. ATP synthase subunit d, mitochondrial. There are 6 proteins decreased:Succinyl-CoA ligase subunit beta, Mannose-6-phosphate isomerase. Protein Serpinb6:Protease inhibitors. Eukaryotic translation initiation factor 2 subunit 1:In eukaryotic cells, protein translation is closely related to the occurrence of tumor development. Anamorsin; Protein Plcxd3:Affectting the lipid metabolism.③ Compared with model group, there are 4 proteins increased in Ququan group: Spetin-11:part of the cell cytokinesis process, transformation of materials inside cells and the process of apoptosis, control the development of embryos. Heat shock cognate 71 kDa protein:maintain the existence and function of cells, improve the resistibility of cells and coordinate with immune function. SAP domain-cotaining ribonucleoprotein:part of the growth and metabolization process of cells and is connected with canceration. There are 4 proteins decreased:Lamin-B1:Affect cell proliferation, migration, an influence of oxidative stress and the signal pathway, has high expression in esophageal cancer, liver cancer tissue. cAMP-dependent protein kinase type Ⅱ-alpha regulatory subunit:mediate the protein phosphorylation process. Secernin-1:part of the cellular function adjustment, and is directly related to diseases such as neural dysfunction. Septin 8 (Predicted):Regulare cell vomit;highly expressed in gastric cancer, prostate cancer and synovial sarcoma.④ Compared with model group, there are 3 proteins increased in non-acupuncture group. Heatshock 70 kDa protein 4:part of the heat shock response, protect the cardiac musle. Alanine-tRNA ligase, cytoplasmic:parts of the synthesis of priteins.2 proteins are decreased. Hsp90 co-chaperone Cdc37:participate in various process of the tumor. Guanine nucleotide-binding protein G(o) subunit alpha:part of the adjustment of the ion channels and signal’s transformation, probably connected with neuroglioma, breast cancer and liver cancer.⑤ Compared with normal group, there are 2 proteins increased in model group: Glycine-tRNA ligase(fragment):control and adjust the genetic expression, suppress the process of cellular apoptosis. Spermatid-associated protein:connected with sterility and infertility.There are 6 protein decreased:Protein disulfide-isomerase A3:parts of the process of cellular signal transduction, promote the folding quality of glycoprotein to be controlled by endoplasmic. Fructose-bisphosphate aldolase C:the enzyme of the glycolysis process, connected with the sugar metabolic enzymes in tumor cells. Biliverdin reductase A:part of the process of phosphorylation, effect the metabolic proteins, proapoptotic molecule cell cycle proteins, signal pathway proteins and expression of gene of the immune factors and oxidative stress factors. Inositol monophosphatasel:parts of GPI transmission process.Phosphoribosyl transferase domain containing 1:connected with the destruction of tumor cell.Conelusion1. Compared with normal Wistar rats, the express of protein with functions of cellular signal transduction, sugar metabolism, synthesis of immune molecules reduced, which turns SHR to have higher risks to be diagnosed with multiple diesases.2. Needling Taichong, Ququan and Chongyang created positive influence physically. It was a result of the multiple targets function of acupuncture. The theory explains the reason that acupoint can improve various symptoms to some extent.3. Needle at non-acupoint shows simple and irregular stimulus to the oblongata.4. The effects of acupoints and the mechanism are all specific.