The Influence Of Ziyin Kidney On Bone-kidney Endocrine Axis Aging Rat Model Based On Klotho/FGF-23 Signaling Pathway

Background: Population aging has become a serious social problem in China,Klotho gene is been thought to be the aging-suppressor genes, together with the FGF-23 receptors, can regulate the homeostasis of calcium, phosphorus and PTH in blood.By effect of nourishing yin and tonifying the kidney on bone-kidney and endocrine axis in order to have certain effects on anti-aging..Objective: Based on klotho/FGF ziyin kidney- 23 signaling pathway research method for the influence of aging rat model of bone-kidney endocrine axis.Methods:Modeling methods:Choose healthy male Wistar rats, 40 weight(200 ±20) g, with random tables divided into groups, model groups, groups of traditional Chinese medicine of nourishing the kidney and Western medicine positive control(vitamin e) groups four groups, each group 10, adaptability of feeding, after 1 week model.Made die method: used D-half lactose induced aging method, will D-half lactose 25 G dissolved Yu 0.9% sodium chloride liquid 500 ml in the, with 500mg/kg abdominal injection dose d(0.01ml/g)give big rat abdominal injection, normal control group abdominal injection,volume of 0.9% sodium chloride injection(0.01ml/g), weekly weighing weight 1 times, according to late times weighing of weight adjustment to drug dose, made die time for 8 week.Medication : Start modeling at the same time irritates the stomach,placebo saline volume groups, model groups, 1 time a day.Set of daily oral administration of traditional Chinese medicine of nourishing the kidney dose of 3.24G/kg, delivery capacity of up to 1ml/100 g.Intragastric administration of positive control of Western medicine(vitamin e) group 0.027g/kg.Calculate weight in rats given each week, and adjust dose based on weight.Index:(1) blood urea nitrogen(Bun), serum creatinine(Scr), blood levels of calcium(Ca2+) and phosphorus(P3+), and 1 day before the blood metabolic cage collection 24 h urine, Urine creatinine(Ucr) measurement and calculation of Creatinine clearance rate(Ccr);(2) detection of Superoxide Dismutase(SOD),Malononitrile(MDA),Nitric Oxide(NO), Nitric Oxide ynthetase(NOS) levels in renal tissue;(3) the rat body weight, diet, hair,activity and there is no death.Statistical methods:Experimental data using SPSS18.0 software for processing, the result( sx ±) that the groups were compared by univariate analysis of variance.Results:1、normal control rats food intake normal, hair light, thick, skin elasticity and lively, not easily removed.After the intraperitoneal injection of d-Galactose, model rats in the control group of emerging color yellow, matte,shed serious bowel movement thin loose stools, diet significantly cuts, slow,reaction ability and activity, genital atrophy, and nature similar to the symptoms of aging rats, suggesting that d-Galactose aging rats showed a significant sign.When modeling 8W, compared with group a group B,significant weight loss, there was a significant difference(P<0.01), after drug therapy, group B, group C and group D with weight increased, but no significant comparison between group C and group D.2、After the use of rats in each group BUN, Scr, Ccr test results show that compared with group a group b, blood BUN, Scr increased significantly;Endogenous creatinine clearance rate was significantly reduced(P<0.01).After treatment with drugs, compared to groups B, C, D and,significant decline in serum BUN, Scr, Ccr increased(P<0.01).Comparison between group C and group D, serum BUN, Scr and slightly decreased, Ccr slightly elevated, but not statistically significant between the two groups(P>0.05).3、Of rats in each group after the administration of blood Ca2+, P3+, ALP test results: B group as compared to group A, ALP, P3+ increase, Ca2+significantly decreased(P<0.01).After drug treatment after the intervention,group C, D Ca2+, P3+, and ALP has significant difference compared with Group B(P<0.01).C, D no difference comparison(P>0.05).4、Detection of FGF-23 level in peripheral blood of rats in each group, the results showed:(1) after the intraperitoneal injection of d-Galactose, FGF-23 level have shown a rising trend in Group b, with a group, there was significant difference(P<0.01).(2) after the treatment, b c, d FGF-23 levels decreased significantly compared and statistically significant(P<0.01), but no significant difference between group c and group d(P>0.05), the two groups was statistically significant compared with group a(P<0.05).5、On the Group big rat kidney organization SOD activity determination,results displayed:(1) made die Hou, made die b group big rat kidney SOD of vitality compared a, group obviously reduced(P<0.01), the treatment group of index greatly adjustable high, and of compared also has difference(P<0.01),description big rat kidney organization SOD activity with aging and declined,and the treatment group has improve its activity of function.(2) c, d can increase activity of SOD, but compared with the normal group, and decrease(P<0.05).6、 On the Group big rat kidney organization c II aldehyde content of determination, results displayed:(1) made die Hou, made module big rat kidney organization MDA of determination more normal group rose, has can than sex(P<0.01), the treatment group of MDA content obviously declined,has can than sex(P<0.01,P<0.05), description big rat aging yihou, kidney organization MDA content significantly increased, and the treatment group are has cut MDA of role.(2) compared with group a, group d was not statistically significant(P>0.05), and group c compared with a group, with comparable(P<0.05).7、On the Group big rat serum NO of detection, results showed that:(1)made die Hou, b group big rat blood NOS of determination obviously increased, and a, group compared has differences sex(P<0.01), the treatment group of determination obviously reduced, and of phase compared has significantly sex differences(P<0.05), description big rat aging Hou, serum in the NOS of content obviously increased, and the treatment group on its has reduced of role.(2) compared with the control group, treatment group had significant statistical difference(P<0.05), d-Galactose can cause increased levels of NOS in aged rats, decline in the treatment group can play a role of NOS, but cannot be restored to a normal level.8、By completing the rat femur bone mineral density testing showed a significantly lower in Group b(P<0.05), no difference compared to group c and group d(P>0.05).After drug therapy, group c and group d, bone mineral density increased significantly as compared with Group b(P<0.05).9 、 By Western blot detection of renal expression of KL, the results indicate that a group is higher than the b group(P<0.01), group c and group d KL protein expression of significant statistical significance compared with Group b(P<0.05).10、By femoral FGF-23 protein levels in the Western blot test, results showed that femoral FGF-23 Group b protein expression level was significantly higher in the Group(P<0.01).After drug therapy, group b, group c and group d with FGF-23 protein levels decreased significantly(P<0.01).11、The rat femoral FGF-23 fluorescence results showed that rats in each group see fluorescent red dye for FGF-23 expression, expression of FGF-23 in Group b a group increased significantly(P<0.05);B, c, d positive Group(P<0.05).To color images showing their relative absorbance of Group b a group showed increased(P<0.05).Group c and d group was significantly decreased compared with Group b(P<0.05), compared with the c group in group d despite the decline but meaningless(P>0.05).12、Using fluorescent quantitative PCR calculation big rat kidney cortex in the of klothom RNA of expression, results displayed: a, group kidney cortex KL protein of performance revealed high had b, and c, and d group, b group with a, group compared greatly declined(P<0.01), c group and d group kl protein of expression and b group compared shang lift(P<0.01), c group and d group zhijian compared no differences sex(P>0.05).13、Nourishing yin and tonifying the kidney cells in renal tissues of rats are the treatment to recover to some extent, observed under the light microscope, have obvious changes compared with the Group;Modules under the electron microscope the Glomerular basement membrane thickening,atrophy characteristic of mesangial matrix increase of vessel blockage symptoms;Degeneration of Glomerular epithelial foot processes widely fusion,swelling of mitochondria and endoplasmic reticulum lumen, nuclear contract changes or even rupture;Under the electron dense deposits, liuwei dihuang pill on ultrastructure of the cells can definitely improve.Conclusions: kidney empty is the foundation of aging, of which the kidney essence deficiency(Yin) for this, so the treatment, when the nourishing kidney Yin(fine) first. Liuwei dihuang pill for nourishing kidney Yin(fine) on behalf of the formula, function, curative effect. Promotes the body’s antioxidant capacity, at the same time maintain normal kidney tissue cells shape profile to achieve; KL protein expression regulation of renal tissue, presumably of nourishing the kidneys through kidney endocrine axis klotho/FGF-23 signaling pathway in bone-in order to improve the effects of aging.