摘要
目的:观察补肾活血方通过调节骨保护素(OPG)/核因子κB活化因子受体(RANK)/核因子κB活化因子受体配体(RANKL)系统对慢性肾脏病(CKD)大鼠血管钙化(VC)的抑制作用,探讨补肾活血方抑制CKD大鼠VC的作用机制。方法:60只SD大鼠随机分为正常组、模型组及中药组,每组20只,模型组与中药组大鼠实验第1-4周给予250mg·kg-1·d-1腺嘌呤混悬液灌胃及含1.8%高磷饲料喂养4周,第5-8周腺嘌呤改为隔日灌胃,中药组同时按55g·kg-1·d-1剂量补肾活血方灌胃治疗,正常组灌胃等量0.9%氯化钠溶液。给药8周后,HE染色观察肾组织变化,茜素红染色观察主动脉钙化结节情况,检测主动脉钙含量及碱性磷酸酶(ALP)活性,腹主动脉取血检测血清钙(Ca)、磷(P)、血肌酐(Scr)、尿素氮(BUN)、全段甲状旁腺激素(i PTH),Western blot法检测主动脉α-平滑肌蛋白(α-SMA)、ALP蛋白的表达,Real-time PCR法检测主动脉OPG、RANKL m RNA表达。结果:HE染色观察提示:中药组较模型组正常肾小球数目增多,肾小管扩张程度降低,炎性细胞数量下降,腺嘌呤结晶减少。茜素红染色观察提示:与正常组比较,模型组主动脉出现钙化结节,广泛红色颗粒物沉着;与模型组比较,中药组钙化结节减少。与正常组比较,模型组主动脉钙含量及ALP活性升高,血P、Scr、BUN、i PTH水平升高,血Ca水平下降(P<0.01),主动脉ALP蛋白表达明显升高、α-SMA蛋白表达明显下降(P<0.01),模型组主动脉OPG m RNA表达下调、RANKL m RNA表达上调(P<0.01);与模型组比较,中药组主动脉钙含量及ALP活性明显降低(P<0.05),血P、Scr、BUN、i PTH水平均下降(P<0.01,P<0.05),血Ca水平升高(P<0.01),中药组主动脉ALP蛋白表达降低、α-SMA蛋白表达升高(P<0.01),且中药组OPG m RNA表达上调(P<0.01),RANKL m RNA表达有下调趋势,但差异无统计学意义。结论:补肾活血方能够改善CKD大鼠肾脏组织学、肾功能、改善钙磷代谢,上调OPG/RANK/RAN
Objective： To explore the inhibiting effect and mechanism of Bushen Huoxue Decoction（BHD） on vascular calcification（VC） in chronic kidney disease（CKD） rats by regulating osteoprotegerin（OPG）/receptor activator of nuclear factorkappa B（RANK）/receptor activator of nuclear factor-kappa B ligand（RANKL） system. Methods： Sixty SD rats were randomly divided into the normal group, model group and BHD group, 20 rats in each group. 250mg/kg adenine suspension by gastrogavage and 1.8% phosphorus content by dietary were executed to molding for 4 weeks. Then, during 5-8 weeks, adenine gavage was given once in 2d; at the same time, the rats in BHD group were given BHD gavage and control group were given normal saline equivalent. Eight weeks later, the histological changes in renal and aorta of rats were observed by HE staining and alizarin red staining, respectively. Calcium level and alkaline phosphatase（ALP） activity of the aorta were detected. The level of calcium（Ca）, phosphorus（P）, creatinine（Scr）, blood urea nitrogen（BUN）, intact parathyroid hormone（i PTH） in serum were detected via abdominal aortic blood. The protein expression level of α-smooth muscle actin（α-SMA） and ALP in aorta were detected by Western blot. The m RNA expression of OPG and RANKL were detected by Real-time PCR. Results： HE staining showed that the number of glomerular improved, compared with the model group, the expansion degree of renal tubular decreased, the number of inflammatory cells decreased, and the crystallization of adenine decreased in the BHD group. Alizarin red staining showed that： compared with the normal group, the calcified nodules were found in the model group with wide red particle deposition in aorta; compared with model group, the calcified nodules decreased in the BHD group. Compared with the normal group, the calcium level and ALP activity of aorta and the serum P, Scr, BUN, i PTH levels significantly increased, while the serum Ca level significantly decreased in mod
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2016年第5期1652-1658,共7页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金资助项目(No.81273747)
中国中医科学院博士研究生创新人才培养基金资助项目(No.CX201506)~~
