| Objective:In this study, rats Yiqibushen side of streptozotocin (STZ)-induced DN, exploreYiqibushen parties kidney of diabetic rats hypoxia inducible factor-1a (HIF-1a), Nox4,p47phox expression influence, and their role in the anti-diabetic rats hypoxic damageaspects of traditional Chinese medicine treatment of diabetes and its complicationsrevealing provide experimental and theoretical basis.Method：diabetic nephropathy rat model was prepared in SPF72male Wistar rats, weighingabout200g, were randomly divided into: normal control group, diabetic nephropathymodel group and the treatment group, diabetic nephropathy model group and treatmentgroup with prior modeling high methionine fed high cholesterol diet for2weeks. All ratswere fasted before modeling can not help but water16h, the diabetes group and treatmentgroup of rats by a single intraperitoneal injection of60mg/kg dose of1%STZ solution,72h after tail vein blood glucose measurements to three consecutive non-fasting bloodglucose>16.67mmol/L, urine output>150%of the original amount of urine, urinaryprotein excretion>30mg/24h molded in urine by standard. Normal control rats wereinjected with a single dose of buffer, the remaining operation with other groups. Treatmentgroup fed with aqueous Yiqibushen parties, and the remaining two groups with salinegavage once daily for12weeks. During the experiment, we observed the general state of rats in each group, and after12weeks, renal tissues of each group were taken for sections,the following HE staining, immunohistochemistry and RT-PCR assay.Results:High cholesterol high methionine diet combined with intraperitoneal injection of asmall dose of streptozotocin-induced diabetic nephropathy model can succeed.1.A general state of the rats of normal control rats eating, drinking, the mental state,daily activities were normal weight with weeks to grow. Performance of diabeticnephropathy model rats as weight loss, polydipsia, polyphagia, polyuria apparentlistlessness, dark color, unresponsive and other common features of diabetic rats, and someappear to reduce back hair, buttocks brown coat, body, tail infection fester, cataracts andother complications, rat stool color depth of thin, strong urine smell was sour. Treatmentgroup had a similar performance in rats, but the extent compared with the model groupreduced.2.HE staining light microscope normal rats showed normal renal structure, noglomerular hypertrophy and tubular structures showed no abnormalities. DN model group,pathological changes in the kidney glomerular volume increased glomerular capillaryendothelial cells and mesangial cell proliferation, basement membrane thickening,mesangial widening, tubular epithelial cell edema. Treatment group edema renal tubularepithelial cells, capillary endothelial cells and mesangial cell proliferation reduced,basement membrane thickening reduced.3.Immunohistochemistry showed that normal rat kidney tubular epithelial cells, renalinterstitial cells have a very small amount of HIF-1a, P47, NOX4expression, almost noexpression in mesangial cells cytoplasm; DN group between renal tubular epithelial cellsand interstitial cells cytoplasm HIF-1a, P47, NOX4showed strong positive expression.Group of pictures brown particles HIF-la, P47, NOX4positive expression, measured usingIPP software and compared its IOD value of each group HIF-la, P47, NOX4expression.Treatment group and diabetes group HIF-la, P47, NOX4expression levels than the controlgroup (P<0.05or P <0.01). Treatment group HIF-la, P47, NOX4expression levels lessthan the diabetic group (P<0.05or P <0.01), higher than the normal group (P>0.05); diabetic group than in the normal group of high expression levels.4.RT-PCR results in the normal control group based on a value of1. Compared withnormal control group, diabetes model group HIF-1amRNA, p47phox mRNA and Nox4mRNA expression levels were significantly increased (P <0.01); compared with diabeticmodel group, treatment group HIF-1amRNA, p47phox mRNA and Nox4the mRNAexpression levels decreased (P <0.05). High expression of traditional Chinese medicinetreatment group compared with the control group (P <0.05).Conclusion:1.Yiqibushen side can improve the general state of experimental diabeticrats, lower blood sugar in diabetic rats, reduce urinary excretion of protein, protect thekidneys.2.After Yiqibushen party intervention can decrease HIF-1amRNA, p47phox mRNAand Nox4mRNA expression levels, inhibition of early renal hypertrophy andhyperfiltration, and improve the renal, thereby reducing kidney damage in diabetic rats,delay the onset of diabetic nephropathy development. |
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