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黄芪水提物对慢性肾功能衰竭模型大鼠的改善作用及其对MAPK信号通路的影响
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| 篇名： | 黄芪水提物对慢性肾功能衰竭模型大鼠的改善作用及其对MAPK信号通路的影响 |
| TITLE： | |
| 摘要： | 目的：考察黄芪水提物对慢性肾功能衰竭（CRF）模型大鼠的改善作用及其对丝裂原活化蛋白激酶（MAPK）信号通路的影响，探讨其可能机制。方法：将雄性SD大鼠随机分为对照组（10只）和造模组（50只），后者灌胃25％腺嘌呤混悬液200 mg/kg（每天1次，连续28 d）以复制CRF模型。造模后，将造模组大鼠随机分为模型组、贝那普利组（阳性对照，2 mg/kg）和黄芪水提物低、中、高剂量组（1.5、3、6 g/kg，按生药量计），每组10只。对照组和模型组大鼠均灌胃等体积生理盐水，各给药组均灌胃相应药物；每天1次，连续28 d。末次给药12 h后，采用比色法检测各组大鼠血清肾功能指标（血肌酐、尿素氮、尿酸）含量，采用酶联免疫吸附测定法检测其血清炎症因子[白细胞介素1β（IL-1β）、IL-6、肿瘤坏死因子α]水平，分别采用羟胺法、可见分光光度法和硫代巴比妥酸法检测各组大鼠肾组织中氧化应激相关指标[超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、丙二醛（MDA）]的活性或含量，分别采用实时聚合酶链反应法、Western blotting法检测其肾组织中凋亡相关因子[Bax、Bcl-2、胱天蛋白酶3（Caspase-3）]mRNA以及MAPK信号通路相关调控蛋白[p38 MAPK、磷酸化p38 MAPK（p-p38 MAPK）、细胞外调节蛋白激酶1/2（ERK1/2）、磷酸化ERK1/2（p-ERK1/2）、c-Jun氨基末端激酶（JNK）、磷酸化JNK（p-JNK）]的表达情况。结果：与对照组比较，模型组大鼠血清肾功能指标含量、炎症因子水平以及肾组织MDA含量、Bax与Bcl-2 mRNA相对表达量的比值（Bax/Bcl-2）、Caspase-3 mRNA的相对表达量、MAPK信号通路相关调控蛋白磷酸化产物（p-p38 MAPK、p-JNK、p-ERK1/2）的相对表达量均显著升高，肾组织SOD、CAT活性均显著降低（P＜0.01）。与模型组比较，各给药组大鼠血清肾功能指标含量、炎症因子水平以及肾组织Bax/Bcl-2、MAPK信号通路相关调控蛋白磷酸化产物的相对表达量，贝那普利组和黄芪水提物中、高剂量组大鼠肾组织MDA含量、Caspase-3 mRNA相对表达量均显著降低；贝那普利组和黄芪水提物中、高剂量组大鼠肾组织SOD、CAT活性均显著升高（P＜0.05或P＜0.01）；且贝那普利组MDA含量、Bax/Bcl-2均显著低于黄芪水提物高剂量组（P＜0.05）。结论：黄芪水提物对CRF模型大鼠具有一定的改善作用，可抑制其炎症反应、氧化应激和细胞凋亡，其作用机制可能与抑制MAPK信号通路相关调控蛋白的表达有关。 |
| ABSTRACT： | OBJECTIVE： To investigate the improvement effects of water extract of Astragalus membranaceus on chronic renal failure （CRF） model rats and its effects on MAPK signaling pathway， and to investigate possible mechanism. METHODS： Male SD rats were randomly divided into control group （10 rats） and model group （50 rats）. CRF model was established by intragastric administration of 25％ Adenine suspension 200 mg/kg （once a day， for consecutive 28 d）. After modeling， modeling group was randomly divided into model group， benazepril group （positive control， 2 mg/kg）， A. membranaceus water extract low-dose， medium-dose and high-dose groups （1.5， 3， 6 g/kg，by crude drug）， with 10 rats in each group. Control group and model group were given constant volume of normal saline intragastrically， and administration groups were given relevant medicine intragastrically， once a day， for consecutive 28 d. Twelve hours after last medication， the contents of serum renal function indexes （serum creatinine， urea nitrogen， uric acid） were determined by colorimetry. ELISA method was used to detect the levels of serum inflammatory factors （IL-1β， IL-6 and TNF-α）. The activity or content of oxidative stress related indexes （SOD， CAT and MDA）in renal tissue of rats were determined by hydroxylamine method， visible spectrophotometry method and thiobarbituric acid method. The mRNA expression of apoptosis-related factors （Bax， Bcl-2， Caspase-3） in renal tissue， MAPK signaling pathway-related regulatory protein p38 MAPK， phosphorylated p38 MAPK（p-p38 MAPK）， ERK1/2， phosphorylated ERK1/2 （p-ERK1/2）， JNK and phosphorylated JNK （p-JNK） were determined by real- time PCR and Western blotting assay. RESULTS： Compared with control group， the contents of renal function indexes and the levels of inflammatory factors in serum， MDA content， mRNA expression ratio of Bax and Bcl-2 （Bax/Bcl-2）， mRNA related expression of Caspase-3， related expression of phosphorylation product of MAPK signaling pathway-related regulatory proteins （p-p38 MAPK， p-JNK， p-ERK1/2） in renal tissue were increased significantly in model group， while the activities of SOD and CAT were decreased significantly in renal tissue （P＜0.01）. Compared with model group， the contents of renal function indexes and the levels of inflammatory factors in serum， Bax/Bcl-2， related expression of phosphorylation product of MAPK signaling pathway-related regulatory proteins in renal tissue were decreased significantly in administration groups as well as MDA content and mRNA related expression of Caspase-3 were decreased significantly in benazepril group and A. membranaceus water extract medium-dose and high-dose groups； the activities of SOD and CAT in renal tissue were increased significantly in benazepril group and A. membranaceus water extract medium-dose and high-dose groups （P＜0.05 or P＜0.01）. MDA content and Bax/Bcl-2 of benazepril group were significantly lower than those of A. membranaceus water extract high-dose group （P＜0.05）. CONCLUSIONS： A. membranaceus water extract has a certain improvement effect on CRF model rats， and inhibit its inflammatory reaction， oxidant stress and cell apoptosis， the mechanism of which may be associated with inhibiting the expression of MAPK signaling pathway-related regulatory proteins. |
| 期刊： | 2019年第30卷第10期 |
| 作者： | 汪卫红，许烨，李志明，远方 |
| AUTHORS： | WANG Weihong，XU Ye，LI Zhiming，YUAN Fang |
| 关键字： | 黄芪水提物；慢性肾功能衰竭；炎症反应；氧化应激；细胞凋亡；MAPK信号通路；调控蛋白；大鼠 |
| KEYWORDS： | Astragalus membranaceus water extract； Chronic renal failure； Inflammation reaction； Oxidant stress； Cell apoptosis； MAPK signaling pathway； Regulatory protein； Rats |
| 阅读数： | 440 次 |
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