Effect Of Electroacupuncture On T Lymphocyte Subsets And TLR4 Signaling Pathway In Prehypertensive Rats

Purpose:The experiment model established by 8% Nacl salt feeding prehypertension rat,then electrically stimulatied Dahl salt-sensitive rats Zusanli(ST36) and Quchi(ST11),and observed Dahl salt-sensitive rat tail artery systolic pressure CD4+ T cells and CD8+ T cells in peripheral blood and TLR4 signaling pathway of Toll-like receptor 4,tumor necrosis factor receptor-associated factor 6, a transcriptional activator protein 1 m RNA in the spleen macrophages.Evaluated affects of acupuncture regulating the immune system to low-grade inflammation prehypertension blood pressure,and to further explore the mechanism and prehypertension and Spleen Qi immunocompromised relationship. Material and method:7-week-old salt-resistant rats(DR),male,10,is set to the normal control group;Dahl 7-weeks-old salt-sensitive rats,male,30.30 7-week-old Dahl salt-sensitive rats(DS)were randomly divided into three groups,each 10,the model group(bundled with every fixed 15 min,not acupuncture),acupuncture group(after the rats were tied bilateral Quchi points, Zusanli point,needle into 3-5mm,connect electrical stimulation of acupuncture device 15 min,the use of density wave,frequency 2HZ,strength to handle the needle tremble for the degree once a day, six times a week), non-acupoint group(rats were tied after bilateral Zusanli beside 1-2mm, the leading edge of the deltoid and biceps in the upper arm junction needle into 3-5mm,connect electrical stimulation of acupuncture device 15 min,the use of density wave,frequency 2HZ,strength to handle the needle tremble for the degree,once a day,six times a week).First,8% Nacl high salt feeding and the use of intelligent non-invasive blood pressure monitoring blood pressure every day,one week after the rise to the level of pre-hypertension,then feeding ordinary diet,continuous electro-acupuncture treatment for 4 weeks,and intelligent non-invasive blood pressure measuring blood pressure weekly,observed changes in rat tail artery systolic pressure until the blood pressure to normal levels,then stoping treatment.After treatment, 2ml blood was collected in each group of rats, comprising acetic acid drawn into tetraacetic acid(EDTA) and salts anticoagulant blood collection tube, point up and down repeatedly to 8-10 times, 6 hours using flow technically machine type handle; the other with 10% chloral hydrate anesthesia(3ml / kg body weight). After the rats were anesthetized rat fixed on the stage, with a 0.5% povidone-iodine disinfection rat abdominal skin, cut the skin and peritoneum, fully exposed to the spleen, the spleen tissue about 50 mg adding 1m L Trizol reagent in the EP tubes at-80℃ refrigerator alternate, total RNA in spleen muscle tissue was extracted,reverse transcribed into c DNA, and then were added to the appropriate primers rat TLR4,TRAF-6 and AP-1 gene to β-actin as an internal control. Detection spleen TLR4,TRAF-6 expression, AP-1m RNA by quantitative PCR.Data use the single factor analysis of variance with the SPSS17.0software,comparing two groups using the LSD method analysis,and results expressed as mean±standard deviation(x±s),P<0.05 was considered significant difference. Results: 1.Observed before acupuncture treatment,the blood pressure of the model group,acupuncture group,and non-acupoint group were higher than the normal group, one week after treatment, compared with the model group,acupuncture group and non-acupoint group of rats systolic blood pressure decreased after 2 weeks of treatment, compared with the model group, acupuncture group and non-acupoint by systolic blood pressure was significantly decreased after 3 weeks of treatment,the rat tail artery systolic pressure continued to decline, treatment 4 weeks later,the acupuncture group,up to a certain non-stable rat tail artery systolic pressure by non-acupoint group.2.Compared with the normal group, model group, acupuncture group CD4+ T lymphocyte percentage decreased significantly, non-acupoint group,the percentage of CD4+ T lymphocytes were also down,but a drop of no acupuncture group significantly, while the normal group and model no significant group differences in the percentage of lymphocytes;compared with the normal group,CD8+ lymphocyte percentages were significantly higher in the model group, CD8+ lymphocyte percentage decreased after acupuncture,non-acupoint group also decreased but still higher than acupuncture group, and the acupuncture group and non-acupoint group CD8+ lymphocyte percentages are lower than the normal group. 3.After four weeks of treatment by acupuncture Toll-like receptor 4(TLR4), compared with the model group,acupuncture group and significantly lower non-acupoint group gene expression of the gene m RNA,and there are significant differences between the four groups;tumor necrosis factor receptor-associated factor 6(TRAF-6)of m RNA compared with the model group,acupuncture group and the normal gene expression was significantly decreased,but there was no significant difference between the model group and non-acupoint group;transcriptional activator protein 1(AP-1)m RNA compared with the model group,acupuncture group and non-acupoint group gene expression was significantly reduced,and there are significant differences among the four groups. Conclusion: 1.Acupuncture on Dahl salt-sensitive rats with blood pressure effect,which make prehypertension blood pressure returned to normal levels. 2.Dysfunction of the immune hypertensive rats pre-existence of lymphocyte activation of inflammatory state, acupuncture can improve the spleen-deficiency-induced low-grade inflammatory prehypertension rats.