作　　者： ; ; ; ; ; ; ;

机构地区： 广州市中医医院

出　　处： 《中国药房》 2019年第16期2193-2199,共7页

摘　　要： 目的:建立健脾益肾丸(JYP)中黄芪甲苷、大黄素、大黄酚的含量测定方法,并探讨JYP对慢性肾衰竭(CRF)模型大鼠钙磷代谢和炎症因子等相关指标的影响。方法:采用高效液相色谱法进行含量测定。黄芪甲苷以及大黄素、大黄酚的色谱柱分别为Agilent Zorbax SB-C18、Agilent TC C18,流动相分别为乙腈-水(36∶64,V/V)、甲醇-0.1%磷酸溶液(75∶25,V/V),检测器分别为蒸发光散射检测器和二极管阵列检测器(后者检测波长为254 nm),柱温分别为30、25℃,流速均为1.0 m L/min,进样量分别为20、10μL。将SD大鼠随机分为正常组、模型组、尿毒清组(1.80 g/kg)和JYP低、中、高剂量组(1.71、3.43、6.85 g/kg),每组10只。除正常组外,其余组大鼠均采用5/6肾切除方法复制CRF模型。造模4个月后,正常组和模型组大鼠均灌胃相应体积水,各给药组大鼠灌胃相应药物,每日1次,连续12周。采用酶联免疫吸附测定法检测大鼠血肌酐(Scr)、尿素氮(BUN)、甲状旁腺激素(PTH)和炎症因子[白细胞介素-6(IL-6)、肿瘤坏死因子α(TNF-α)]含量,采用甲基百里香酚蓝比色法和磷钼酸法检测其血钙、血磷含量,采用Pearson检验考察炎症因子与钙磷代谢相关指标(血钙、血磷、PTH)的相关性。结果:黄芪甲苷、大黄素、大黄酚检测质量浓度的线性范围分别为54.537~381.759、2.960~20.720、6.318~44.223μg/m L(r>0.999),定量限分别为0.010、0.288、0.216μg/mL,检测限分别为0.003、0.096、0.072μg/mL,精密度、重复性、稳定性试验的RSD均小于3.0%,加样回收率为97.18%~102.33%(RSD<3%,n=9)。造模后(给药前),模型组和各给药组大鼠血清Scr、BUN含量均较正常组显著升高(P<0.01);给药后,各给药组大鼠上述指标均较模型组和同组给药前显著降低(P<0.01)。与正常组比较,模型组大鼠血钙含量显著降低,IL-6、TNF-α含量均显著升高(P<0.01);与模型组比较,JYP中、高剂量组大鼠血钙含量均显著升高,尿 OBJECTIVE:To establish the method for the content determination of astragaloside Ⅳ,emodin and chrysophanol in Jianpi yishen pills(JYP)and to investigate the effects of JYP on calcium,phosphorus metabolism and inflammatory factors in chronic renal failure(CRF)model rats. METHODS:HPLC method was adopted. The determination of astragaloside Ⅳ,emodin and chrysophanol was perform on Agilent Zorbax SB-C18,Agilent TC C18 column, respectively;mobile phase consisted of acetonitrile-water(36 ∶ 64,V/V) and methanol-0.1% phosphoric acid solution(75 ∶ 25,V/V);the detectors were evaporative light-scattering detector and diode-array detector(detection wavelength of 254 nm);the column temperatures were set at 30 ℃and25 ℃ at the flow rate of 1.0 m L/min;the sample sizes were 20 and 10 μL. SD rats were randomly divided into normal group,model group,Niaoduqing group(1.80 g/kg)and JYP low-dose,medium-dose and high-dose groups(1.71,3.43,6.85 g/kg),with10 rats in each group. Except for normal group,CRF model of other groups were established by 5/6 nephrectomy in other groups.Four months after modeling,normal group and model group were given constant volume of water intragastrically;administration groups were given relevant medicine intragastrically,once a day,for consecutive 12 weeks. The levels of serum creatinine(Scr),urea nitrogen(BUN),parathyroid hormone(PTH)and inflammatory factors(IL-6,TNF-α)were measured by ELISA. Methyl thymol blue colorimetric method and phosphomolybdic acid method were used to detect the contents of blood calcium and phosphorus. Correlation of inflammatory factors with related calcium and phosphorus metabolism indexes(blood calcium,blood phosphorus,PTH)were investigated with Pearson assay. RESULTS:The linear range of astragaloside Ⅳ,emodin and chrysophanol were 54.537-381.759,2.960-20.720,6.318-44.223 μ g/mL,respectively. The limits of quantitation were 0.010,0.288,0.216 μg/mL;the limits of detection were 0.003,0.096,0.072 μg/mL. RSDs of precision,reproducibility and stability tests were al

关 键 词： 健脾益肾丸 黄芪甲苷 大黄素 大黄酚 含量测定 高效液相色谱法 慢性肾衰竭 钙磷代谢 炎症因子 大鼠

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