芪地糖肾颗粒对糖尿病大鼠认知功能的影响及机制

Effects and Mechanismof QidiTangshen Granules on Cognitive Functionof Rats withDiabetes

目的 探讨芪地糖肾颗粒对糖尿病大鼠认知功能的影响及作用机制。方法 将40只大鼠随机分为对照组、模型组、吡拉西坦组和芪地糖肾颗粒组,每组各10只。对照组正常进食不造模,模型组、吡拉西坦组和芪地糖肾颗粒组建立2型糖尿病模型。造模后吡拉西坦组灌胃0.48 g/kg吡拉西坦,芪地糖肾颗粒组灌胃5.24 g/kg芪地糖肾颗粒,对照组和模型组大鼠灌胃等量生理盐水,均治疗12周。药物干预后检测各组大鼠空腹血糖、胰岛素水平,记录大鼠体重,采用Morris水迷宫实验检测认知功能,Western-blot检测海马组织内质网应激指标[葡萄糖调节蛋白78(Glucose regulated protein78kD,GRP78)、C/EBP同源蛋白(C/EBP homologous protein, CHOP)、磷酸化蛋白激酶R样内质网激酶(p-PERK)、磷酸化肌醇依赖酶1α(p-IRE1α)、转录激活因子(ATF)]和NOD样受体家族3(NLRP3)炎症小体信号通路相关指标[硫氧还蛋白互作蛋白(Thioredoxin-interacting protein, TXNIP)、凋亡相关斑点样蛋白(Apoptosis-associated speck-like protein containing a CARD,ASC)、NLRP3、白细胞介素-1β裂解片段(cleaved IL-1β)]蛋白表达水平,酶联免疫吸附试验(Enzyme linked immunosorbent assay, ELISA)检测海马组织中炎症因子[白细胞介素1β(Interleukin-1β,IL-1β)、白细胞介素6(Interleukin-6,IL-6)和肿瘤坏死因子α(Tumor necrosis factor, TNF-α)]蛋白表达水平。结果 与对照组比较,模型组体重降低(P<0.05),给药前后空腹血糖升高(P<0.05),胰岛素水平降低(P<0.05)。吡拉西坦组与模型组的体重、空腹血糖和胰岛素水平比较无统计学差异(P>0.05)。与模型组比较,芪地糖肾颗粒组体重增高(P<0.05),给药后空腹血糖降低(P<0.05),胰岛素水平升高(P<0.05)。与对照组比较,模型组训练第5天时逃避潜伏期增高(P<0.05),跨平台次数减少(P<0.05)。与模型组比较,吡拉西坦组和芪地糖肾颗粒组训练第5天时逃避潜伏期降低(P<0.05),跨平台次数增加(P<0.05)。吡拉西坦组和芪地糖肾颗粒组逃避潜伏期和跨平台次数比较,差异无统计学意义(P>0.05)。与对照组比较,模型组GRP78、CHOP、p-PERK、p-IRE1α、ATF、TXNIP、ASC、NLRP3、Cleaved IL-1β、IL-1β、IL-6和TNF-α蛋白表达水平升高(P<0.05)。与模型组比较,吡拉西坦组和芪地糖肾颗粒组各蛋白表达水平均明显降低(P<0.05)。吡拉西坦组和芪地糖肾颗粒组海马组织内质网应激指标、NLRP3炎症小体信号通路指标、炎症因子水平比较,差异均无统计学意义(P>0.05)。结论 芪地糖肾颗粒能够通过抑制内质网应激和NLRP3炎症小体信号通路而改善糖尿病大鼠认知功能损伤。

Objective To explore the effects and underlying mechanism of QidiTangshen Granules(QDTS)on the cognitive function of rats with diabetes.Methods Forty rats were randomly divided into a control group, a model group, a piracetam group, and a QDTS group, with 10 rats in each group.Rats in the control group were fed on a normal diet.Thetype 2 diabetes model was induced in rats except for those in the control group.Rats in the piracetam group andthe QDTS group were administered with 0.48 g/kg piracetam and 5.24 g/kg QDTS,respectively, while those in the control group and the model group with an equal volume of normal saline by gavage.After 12 weeks of drug intervention, the fasting blood glucose and insulin levels were detected and the body weight was recorded.Morris water maze test was used to detect cognitive function.Westernblot was used to detect the endoplasmic reticulum stress indicators[glucose-regulated protein 78(GRP78),C/EBP homologous protein(CHOP),phosphorylated protein kinase R-like endoplasmic reticulum kinase(p-PERK),phosphorylated inositol requiring enzyme-1α(p-IRE1α),and activating transcription factor(ATF)]in the hippocampus and related indicators in the NOD-like receptor protein 3(NLRP3)inflammasome signaling pathway[thioredoxin-interacting protein(TXNIP),apoptosis-associated speck-like protein containing a CARD(ASC),NLRP3,and cleaved interleukin(IL)-1β].Enzyme-linked immunosorbent assay(ELISA)was used to detect the protein expression levels of IL-1β,IL-6,and tumor necrosis factor(TNF)-α in the hippocampus.Results Compared with the control group, the model group showeddeceased body weight(P<0.05),increased fasting blood glucosebefore and after administration(P<0.05),and reduced insulin level(P<0.05).There was no difference in body weight, fasting blood glucose, and insulin levels between the piracetam group and the model group(P>0.05).Compared with the model group, the QDTS group showed increased body weight(P<0.05),decreased fasting blood glucoseafter administration(P<0.05),and elevated insulin level(P<0.05).Compared with the control group, the model group showed prolonged escape latency on the 5 th day of training(P<0.05)and reduced cross-platform times(P<0.05).Compared with the model group, the piracetam group and the QDTS group showed shortened escape latency on the 5 th day of training(P<0.05)and increased cross-platform times(P<0.05).There was no difference in escape latency and cross-platform times between the piracetam group and the QDTS group(P>0.05).Compared with the control group, the model group showed higher protein expressionof GRP78,CHOP,p-PERK,p-IRE1α,ATF,TXNIP,ASC,NLRP3,cleaved IL-1β,IL-1β,IL-6,and TNF-α(P<0.05),and the above-mentioned protein expression levels in the piracetam group and the QDTS group were lower than those in the model group(P<0.05).There was no difference in the levels of endoplasmic reticulum stress indicators, related indicators in the NLRP3 inflammasome signaling pathway, and inflammatory factors in the hippocampus between the piracetam group and the QDTS group(P>0.05).Conclusion QDTS can improve the cognitive function impairment of rats with diabetes by inhibiting the endoplasmic reticulum stress and the NLRP3 inflammasome signaling pathway.