Effects Of Twist And Diarrhea On NO, ET-1 And TGF-β1 In Renal Tissue Of Spontaneously Hypertensive Rats

ObjectiveTo explore the different effects of reinforcing and reducing methods by twirling and rotating the needle on prevention for renal damage of SHR,the blood pressure in SHR,endothelin-l,the level of NO,TGF-β1 were measured by different biochemistry methods.Observingthe influence of kidney and adrenal gland tissue morphology change of SHR aims to reveal whether acupuncture has a good effect on preventingrenal damage of SHR.And further research from the level of vascular endothelial function intervention mechanism,Which can also helps to reveal the mechanism in molecular level,evaluatingcurative effects of acupuncrure,providing scientific and objective evidence for different effects of reinforcing and reducing methods by twirling and rotating the needle,providing basic experiment basis for the further study.MethodSixty male SHR of 12 weeks’old were randomly divided into five groups:group (A) with twirling reinforcing method, group (B) with twirling reducing method,group (C) with twirling reinforcingand reducing method,group (D) without manipulation, model group (E), and a normal Wistar rats of the same age with SHR as group (F) without acupuncture. Group A、B、 C、D、E are all treated on single "Taichong" point (LR3) every day,ten minutes every time, rest one day every six days. Before the experiment process, measuring the blood pressure value 3 times, then taking the average value is blood pressure valuebefore the acupuncture.Every six days after the acupuncture treatment began,with noninvasive blood pressure meter measuring quiet, awake rats tail pressure,each group was measured four times, then taking the average is blood pressure after the experiment.28 days later, observing the kidneys and adrenal pathology change. Application of nitrate enzymatic determination of NO in SHR kidney tissue, chemical colorimetry to determine NOS; detecting the expression of TGF-β1 and ET-1,in SHR kidney tissues by immunohistochemical method.Results1. The tissue morphology observationGroup F renal unit and the small arteries in the renal cortex structure are basic normal. Group A, group C, group D all have different degree of glomerular mesangial matrix proliferation, capillary basement membrane thickening, mesangial cell proliferation, visible goal small artery wall thickening, luminal stenosis or obstruction. A small number of visible glomerular sclerosis of focal, renal tubular turbidity is swollen, granular degeneration, irregular thickening of the small jaasiel membrane, interstitial inflammatory cell infiltration, increased extracellular matrix (ECM) and deposition, renal interstitial fibrosis; Group E progression of hypertension, renal tubular lesions aggravated; Group B basic normal blood vessels, basic normal glomerulus F, B group of adrenal weight without or trace increase; The rest of the group increased significantly, with the most obvious group E. A, C, D, E group kidney weight is F, among them the most significant group E, while group B was not significantly reduced.2.The changes of systolic pressure and diastolic pressure of each groupBefore acupuncture, blank group (F) systolic and diastolic blood pressure with A, B, C, D, E group are different (P<0.05).Systolic and diastolic blood pressure of normal blank group is lower than the rest of the five SHR groups.A, B, C, D, E group has no difference between the systolic and diastolic blood pressure(P>0.05). After experimental treatment,group F systolic blood pressure compared with before treatment, no difference (p>0.05), and diastolic blood pressure has increased (P<0.05); Group E systolic and diastolic blood pressure compared with the experiment before all have increased, compared with the experiment before having obvious difference (P<0.05), as the growth of the SHR weeks, blood vessel elasticity decreased, model group blood pressure is on the rise; Compared with group A, C, D group E also has difference (P<0.05); A, C, D group rats before and after treatment has no obvious systolic and diastolic blood pressure change (P>0.05); Group B after treatment of rats is significantly lower in systolic and diastolic blood pressure,there isa significant difference compared with before treatment (P<0.05),compared with A,C,D group has significant difference (P<0.05)compared with group E and F has extremely significant difference (P<0.01).3.The expression of NO and NOS in renal tissue of each groupAfter the acupuncture treatment,the NO content in group E and group F with obvious difference (P<0.05). A,C,D has significant difference between groups (P>0.05),group B compared with A,C,E,D,there is significant differences (all P<0.05). After acupuncture treatment,A,C,D,E group of NOS content in kidney tissue of with F group there is significant differences (P<0.05). The expression level of E group is obviously lower than that of group B (P<0.05); The expression level of group B is obviously higher than that of A, C, D group (P<0.05). Group E and group F have obvious difference (P<0.05).4. the expression of TGF-β1 in renal tissue of each groupAfter the acupuncture treatment,TGF-β1 in the renal tubules in group B, F no or only trace tan particles,TGF-β1 no expression or very weak expression; In group A, C, D of glomemlar mesangial cells, epithelial cells of the renal capsule, plasma particles contains A small amount of tan,TGF-P 1 has a small amount of expressions; Expressed in the malpighian tube significantly;TGF-β1 expression significantly increased in group E.TGF-β1 in the expression of renal tubule and the progress of hypertension is positively correlated (P<0.05).TGF-β1 expression group E and group F are more obvious different (P<0.05). A, C, D, E has no significant difference between groups (P>0.05).Group B compared with A, C, E, D, there is significant differences (all P<0.05).There exists obvious differences in group B compared with group F (P<0.05). With the expression level of group B is lower than that in group A,C,E,D(P<0.05).5. the expression ofET-1 in renal tissue of each groupSix different concentration ofET-1between group (p<0.05), group F the lowest concentration ofET-1, The concentration ofET-1 from high, in what order group E>C>A>D B>F.The higher the concentration of ET-1, the heavier the degree of kidney damage; The lower the concentration, the lighter damage degree. Group B concentrations of endothelin-1 is lower than in group A; Acupuncture has an effect on SHR kidney tissue concentration of ET-1, twisting reducing method reduces the most obvious, that twisting reducing method of SHR kidney plays a protective role.After acupuncture treatment, ET-1 expression group E and group F are more obvious different (P<0.05). A, C, D has no significant difference between groups (P>0.05), group B compared with A, C, D have significant differences (all P<0.05).There exists obvious differences in group B compared with group F (P<0.05). With the expression level of group B was lower than that in group A, C, E, D (P<0.05).ConclusionSpontaneously hypertensive rat model is applied in this experiment, by observing the structure morphologies of the adrenal glands, kidneys, detection of SHR rats blood pressure change, NO and ET-1, the change of TGF-β1, the results showed that acupuncture twisting reducing method can make effectively to reduce blood pressure in spontaneously hypertensive rats, and close to normal. Spontaneously hypertensive rats kidney and adrenal gland in pathological injury, acupuncture twisting reducing method of SHR rats kidney pathology organization structure has certain repair effect.After treatment with acupuncture twisting reducing method, kidney tissues and the decrease of the TGF-β land ET-1,NO rising, acupuncture twisting reducing method of spontaneously hypertensive rats blood pressure adjustment is a comprehensive adjustment in the central and peripheral NO and ET-1 and TGF-β1 content, by reducing the pressure of blood flow in vascular endothelial cells and smooth muscle mechanical damage, improve endothelial function, reduce the TGF-31, the secretion of ET-1, so as to reduce the incidence of hypertensive renal damage. Increase the content of NO and NOS in spontaneously hypertensive rats, activating guanylate cyclase acid, lower the level of intracellular ca2+, smooth muscle relaxation, finally achieve the purpose of lowering blood pressure, reduce high blood pressure on renal damage.