Study On The Effect And Mechanism Of Oxidative Stress Of Acupuncture With Fuzheng Huoxue For Hypertensive Renal Damage

Objective:To observe the effects of acupuncture on spontaneously hypertensive rats?SHR?with the caudal artery systolic blood pressure,proteinuria,renal function and renal pathological changes and then explore the mechanism of oxidative stress of acupuncture on hypertensive renal damage.Methods:SHR Rats aged 16 weeks were randomly divided into model group,losartan group,BL-23?Shenshu?acupuncture group,BL-17?Geshu?acupuncture group and BL-23+BL-17?shenshu+geshu?acupuncture group.Wistar Kyoto Rats?WKY?aged the same week were used as the normal group,with 10 Rats in each group.The normal group,model group,BL-23 acupuncture group,BL-17acupuncture group,BL-23+BL-17 acupuncture group were all perfused with Physiological saline daily,the losartan group was intragastrically administered with losartan once a day,while the BL-23,the BL-17 and the BL-23+the BL-17 group were treated with acupuncture according to the different acupoint every other day,for 12 weeks.The caudal artery systolic blood pressure of rats was monitor with Noninvasive sphygmomanometer before intervention and at the weekend of 4,8 and12,biochemical method was used to detect urinary protein and renal function of rats.ELISA was used to detect the expression of urinary microproteins and the serum anti-aging gene klotho,the urine klotho.Immunohistochemistry,Western Blot and rt-pcr were used to detect the expression of klotho m RNA and protein gray level in the kidney of each group.TBA method was used to detect the content of malondialdehyde?MDA?in renal tissue,Reactive oxygen species?ROS?activity was detected by chemical fluorescenceand and determination of superoxide dismutase?SOD?activity was by xanthine oxidase method.Pathological changes of HE,PAS and MASSON staining of rat kidney tissues were observed under light microscope,and ultrastructural changes of rat kidney were observed under transmission electron microscope.Results:?1?Caudal artery systolic blood pressure:at the end of the 4th,8th and 12th week of intervention,the caudal artery systolic blood pressure in both the losartan group and the acupuncture group was lower than that in the model group?P<0.01?.At the end of 12th week,the systolic blood pressure of losartan and acupuncture groups was significantly lower than that before intervention?P<0.01?.?2?24h urine protein,urine series microprotein,renal function:at the end of 8 weeks of intervention,24 urinary protein levels of rats in the losartan group,BL-23 group,BL-23+BL-17 group all decreased to different degrees compared with the model group?P<0.05,P<0.01?.At the end of 12th week,the urine protein for 24h of losartan and each acupuncture group was lower than that of the model group?P<0.05,P<0.01?.Compared with this group at the end of 4th week,the urine protein of rats in BL-23+BL-17 group decreased significantly at the end of 12th week?P<0.05?.The serial trace proteins in urine of rats in losartan group and acupuncture group were significantly lower than those in model group?P<0.01?.There was no significant difference between BL-23 group and losartan group in?₁-MG,m ALb and urinary Ig G?P>0.05?.Scr,BUN and UA in the losartan group,BL-23 group,and BL-23+BL-17 group were lower than those in the model group?P<0.05,P<0.01?,while the contents of Scr and UA in the BL-17 group were lower than those in the model group?P<0.05,P<0.01?.?3?Serum klotho,urine klotho and renal tissue klotho:after the intervention,serum,urine klotho in rats of losartan group and each acupuncture group was significantly increased compared with that in the model group?P<0.05,P<0.01?,and there was no significant difference in expression ofserum klotho between BL-23 group and losartan group?P>0.05?.The expressions of klotho m RNA and protein grayscale in renal tissues of losartan group and each acupuncture group were higher than those of model group?P<0.05,P<0.01?.There was no significant difference in the expression of klotho protein between the acupuncture group and losartan group?P>0.05?.?4?Renal tissue MDA?ROS?SOD:at the end of the 12th week of intervention,ROS activity and MDA content in the losartan group and acupuncture group were significantly lower than those in the model group?P<0.01?,while SOD activity was significantly increased?P<0.01?.?5?Renal pathology:losartan and acupuncture intervention can improve renal tubular disorder,small tube atrophy,stenosis,thickening of capillary wall,protein tube type and cell tube type,and pathological changes of kidney under light microscope;Destruction of the lobular structure of the globules,obscuration of the capillary sputum structure,vascular endothelial cell proliferation,fusion and thickening of the basement membrane,collapse of the foot process,disordered arrangement,and other ultrastructural damage of the kidney.Conclusion:In this study,SHR rats were selected as the models of hypertensive renal damage.Intervention of model rats with BL-23 and BL-17 by acupuncture,It indicated that the acupuncture with Fuzheng Huoxue could reduce the systolic blood pressure,24h urine protein and urine micro-protein of SHR model rats,and improve renal function and pathological morphology of the kidney in SHR rats.acupuncture prescription of strengthening and promoting blood circulation could improve the changes of tail artery systolic blood pressure,24h urinary protein,series of microproteins,renal function and pathological morphology of the kidney in SHR rats.The mechanism may be related to the regulation of klotho expression by acupuncture,reduction of MDA content and ROS activity in rat kidney tissue,and enhancement of antioxidant enzyme SOD activity,thereby inhibiting renal oxidative stress damage caused by hypertension.The BL-23 group was superior to the BL-17 group and the BL-23+BL-17 group in reducing the urinary transferrin?TRF?,urinary Ig G,renal tissue ROS,increasing the expression of serum,urine klotho protein and renal tissue klotho m RNA of SHR rats.