糖脂清对糖尿病大鼠海马内质网应激相关因子GRP78、CHOP、Caspase-12表达的影响
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摘要
目的通过观察糖脂清对糖尿病大鼠海马组织的内质网应激相关因子GPR78、CHOP、Caspase-12的影响,探讨其在改善糖尿病大鼠认知功能的作用机制。方法采用高糖高脂饲料联合小剂量链脲佐菌素腹腔注射复制2型糖尿病大鼠模型,并给予糖脂清高、中、低剂量组干预8周后,采用ELISA法检测血清空腹葡萄糖(FPG)、空腹胰岛素(FINS),计算稳态模型的胰岛素抵抗指数(HOMA-IR)、胰岛素敏感指数(ISI);qPCR和Western blot法检测海马组织内质网应激相关因子GPR78、CHOP、Caspase-12的mRNA及蛋白表达水平。结果糖脂清各剂量组均能够显著降低FPG及HOMA-IR(P<0.01),除低剂量组外,糖脂清均可显著提高ISI(P<0.01),3组FINS水平无显著性差异(P>0.05)。与模型组比较,糖脂清低、中、高3个剂量组GRP78mRNA和蛋白相对表达均显著增加,CHOP、Caspase-12mRNA表达均减少(P<0.01),中、高剂量组CHOP、Caspase-12蛋白表达显著减少(P<0.01)。结论糖脂清能够控制血糖,改善胰岛素抵抗,增加胰岛素敏感性,同时能够调节海马内质网应激相关因子GRP78、CHOP、Caspase-12mRNA和蛋白的表达。
OBJECTIVE To observe the effect of Tangzhiqing on the endoplasmic reticulum stress related factors GPR78,CHOP and Caspase-12 in hippocampus of diabetic rats,and to explore its mechanism of improving cognitive function in diabetic rats.METHODS The model of type 2 diabetes mellitus rats was established by intraperitoneal injection of low-dose of streptozotocin combined with high-sugar and high-fat diet,After 8 weeks of intervention with high,medium and low-dose of Tangzhiqing,serum fasting glucose(FPG)and fasting insulin(FINS)were detected by ELISA,the index of insulin resistance(HOMA-IR)and insulin sensitivity index(ISI)were calculated;the expression levels of endoplasmic reticulum stress related genes GPR78,CHOP and Caspase-12 mRNA and protein in hippocampus were detected by qPCR and Western blot assay.RESULTS Different dosages of Tangzhiqing significantly decreased FPG and HOMA-IR(P<0.01);except the low dose group,Tangzhiqing significantly improved ISI(P<0.01);no significant difference existed in FPG level among the 3 groups(P>0.05).Compared to the model group,the relative expression of GRP78 mRNA and protein in the 3 groups of Tangzhiqing increased significantly,and the expression of CHOP and Caspase-12 mRNA in these 3 groups decreased(P<0.01).CHOP and Caspase-12 in middle and high dose group decreased significantly(P<0.01).CONCLUSION Tangzhiqing can control blood sugar,improve insulin resistance,increase insulin sensitivity and regulate the expression of GRP78,CHOP,Caspase-12 in hippocampus.
OBJECTIVE To observe the effect of Tangzhiqing on the endoplasmic reticulum stress related factors GPR78,CHOP and Caspase-12 in hippocampus of diabetic rats,and to explore its mechanism of improving cognitive function in diabetic rats.METHODS The model of type 2 diabetes mellitus rats was established by intraperitoneal injection of low-dose of streptozotocin combined with high-sugar and high-fat diet,After 8 weeks of intervention with high,medium and low-dose of Tangzhiqing,serum fasting glucose(FPG)and fasting insulin(FINS)were detected by ELISA,the index of insulin resistance(HOMA-IR)and insulin sensitivity index(ISI)were calculated;the expression levels of endoplasmic reticulum stress related genes GPR78,CHOP and Caspase-12 mRNA and protein in hippocampus were detected by qPCR and Western blot assay.RESULTS Different dosages of Tangzhiqing significantly decreased FPG and HOMA-IR(P<0.01);except the low dose group,Tangzhiqing significantly improved ISI(P<0.01);no significant difference existed in FPG level among the 3 groups(P>0.05).Compared to the model group,the relative expression of GRP78 mRNA and protein in the 3 groups of Tangzhiqing increased significantly,and the expression of CHOP and Caspase-12 mRNA in these 3 groups decreased(P<0.01).CHOP and Caspase-12 in middle and high dose group decreased significantly(P<0.01).CONCLUSION Tangzhiqing can control blood sugar,improve insulin resistance,increase insulin sensitivity and regulate the expression of GRP78,CHOP,Caspase-12 in hippocampus.
引文
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[16]梁丽娜.滋补脾阴方药对脾阴虚糖尿病脑病大鼠海马内质网应激影响的研究[D].大连:大连医科大学,2009.
[2]VAN DEN BERG E,KLOPPENBORG RP,KESSELS RP,et al.Type 2diabetes mellitus,hypertension,dyslipidemia and obesity:A systematic comparison of their impact on cognition[J].Biochim Biophys Acta,2009,1792(5):470-481.
[3]GUDALA K,BANSAL D,SCHIFANO F,et al.Diabetes mellitus and risk of dementia:A meta-analysis of prospective observational studies[J].J Diabetes Investig,2013,4(6):640-650.
[4]STRACHAN MW,REYNOLDS RM,MARIONI RE,et al.Cognitive function,dementia and type 2diabetes mellitus in the elderly[J].Nat Rev Endocrinol,2011,7(2):108-114.
[5]XU W,CARACCIOLO B,WANG HX,et al.Accelerated progression from mild cognitive impairment to dementia in people with diabetes[J].Diabetes,2010,59(11):2928-2935.
[6]杨晓青,王旭,邵鑫.糖脂清方通过保护海马神经元对提高2型糖尿病大鼠学习记忆能力的影响[J].中国实验方剂学杂志,2018,24(10):360-365.
[7]蒯美玉,马志,孙鑫,等.桑酮碱改善2型糖尿病大鼠的糖脂代谢[J].中成药,2017,39(1):21-26.
[8]袁有才,问莉娜,曹旸,等.糖尿病认知功能障碍的中医病因病机研究进展[J].光明中医,2016,31(24):3687-3690.
[9]SUN FC,WEI S,LI CW,et al.Localization of GRP78to mitochondria under the unfolded protein response[J].Biochem J,2006,396(1):31-39.
[10]CHEN X,LI M,CHEN D,et al.Autophagy induced by calcium phosphate precipitates involves endoplasmic reticulum membranes in autophagosome biogenesis[J].PLoS ONE,2012,7(12):e52347.
[11]BOUMAN L,SCHLIERF A,LUTZ AK,et al.Parkin is transcriptionally regulated by ATF4:evidence for an interconnection between mitochondrial stress and ER stress[J].Cell Death Differ,2011,18(5):769-782.
[12]PUTHALAKATH H,O'REILLY LA,GUNN P,et al.ER stress triggers apoptosis by activating BH3-only protein Bim[J].Cell,2007,129(7):1337-1349.
[13]ROY S,SHAROM JR,HOUDE C,et al.Confinement of caspase-12proteolytic activity to autoprocessing[J].Proc Natl Acad Sci USA,2008,105(11):4133-4138.
[14]WENG S,ZHU X,JIN Y,et al.Protective effect of erythropoietin on myocardial infarction in rats by inhibition of caspase-12expression[J].Exp Ther Med,2011,2(5):833-836.
[15]CHEN YH,WU XD,YAO ST,et al.Calcineurin is involved in cardioprotection induced by ischemic postconditioning through attenuating endoplasmic reticulum stress[J].Chin Med J(Engl),2011,124(20):3334-3340.
[16]梁丽娜.滋补脾阴方药对脾阴虚糖尿病脑病大鼠海马内质网应激影响的研究[D].大连:大连医科大学,2009.