摘要
目的探讨黄芪甲苷通过Toll样受体4(TLR4)/核因子κB(NF-κB)信号通路对肾脏缺血再灌注大鼠的影响。方法30只健康雄性SD大鼠,随机分为假手术组、缺血再灌注组、黄芪甲苷组。黄芪甲苷组灌胃剂量为40 mg/kg,假手术组和缺血再灌注组灌胃等量生理盐水,连续14 d。第15天,肾脏缺血60 min再灌注24 h,检测大鼠血清尿素氮(BUN)和肌酐(Cr)水平,苏木精-伊红(HE)法检测大鼠肾组织病理学改变,Real-time PCR法检测肾组织TLR4、NF-κB和肿瘤坏死因子α(TNF-α)的mRNA表达,蛋白质印迹法检测肾组织TLR4、NF-κB和TNF-α的蛋白表达。结果缺血再灌注组大鼠血清BUN水平为(18.84±2.33)mmol/L,高于假手术组的(5.31±0.56)mmol/L,t=17.852,P<0.001;缺血再灌注组大鼠血清Cr水平为(169.03±15.37)μmol/L,高于假手术组的(38.15±3.76)μmol/L,t=26.163,P<0.001;黄芪甲苷组大鼠血清BUN为(9.63±1.74)mmol/L,Cr为(95.41±5.83)μmol/L,与缺血再灌注组比较,BUN(t=10.021,P<0.001)和Cr(t=14.158,P<0.001)水平均明显降低。与假手术组比较,缺血再灌注组大鼠肾脏病理损伤严重,肾组织TLR4(t=23.541,P<0.001)、NF-κB(t=28.613,P<0.001)和TNF-α(t=16.302,P<0.001)的mRNA表达明显上调,肾组织TLR4(t=3.718,P=0.002)、NF-κB(t=5.892,P<0.001)和TNF-α(t=9.137,P<0.001)的蛋白表达明显上调;通过黄芪甲苷治疗后,肾脏病理损伤减轻,肾组织TLR4(t=12.759,P<0.001)、NF-κB(t=14.442,P<0.001)和TNF-α(t=7.156,P<0.001)的mRNA表达明显下调,肾组织TLR4(t=3.782,P=0.001)、NF-κB(t=3.102,P=0.006)和TNF-α(t=3.023,P=0.007)的蛋白表达明显降低。结论黄芪甲苷可降低缺血再灌注大鼠炎症因子的表达,减轻肾脏损伤,其机制可能与抑制TLR4/NF-κB信号通路有关。
Objective To investigate the effect of astragalosideⅣon renal ischemia-reperfusion in rats through toll-like receptor 4(TLR4)/nuclear factor kappa-B(NF-κB)signal pathway.Methods Thirty healthy male SD rats were randomly divided into sham operation group,ischemia-reperfusion group and astragalosideⅣgroup.The gavage dose of astragalosideⅣgroup was 40 mg/kg.The sham operation group and ischemia-reperfusion group were gavaged with the same amount of normal saline for 14 days.The 15 th day,after renal ischemia for 60 min and reperfusion for 24 h,the levels of blood urea nitrogen(BUN)and creatinine(Cr)were detected,and the pathological changes of renal tissue were detected,and the mRNA expression of TLR4,NF-κB and tumor necrosis factor-α(TNF-α)in renal tissue were detected by real-time PCR,and the protein expression of TLR4,NF-κB and TNF-αin renal tissue were detected by western blotting.Results The serum BUN level in ischemia-reperfusion group was(18.84±2.33)mmol/L,higher than that in sham operation group(5.31±0.56)mmol/L,t=17.852,P<0.001.The serum Cr level in ischemia-reperfusion group was(169.03±15.37)μmol/L,higher than that of sham operation group(38.15±3.76)μmol/L,t=26.163,P<0.001.In astragalosideⅣgroup,the serum BUN was(9.63±1.74)mmol/L,and the Cr was(95.41±5.83)μmol/L,compared with the ischemia-reperfusion group,the levels of BUN(t=10.021,P<0.001)and Cr(t=14.158,P<0.001)decreased significantly.Compared with the sham operation group,the renal pathological injury in the ischemia-reperfusion group was serious,and the mRNA expression of the renal tissue TLR4(t=23.541,P<0.001)and NF-κB(t=28.613,P<0.001)and TNF-α(t=16.302,P<0.001)were significantly up-regulated,and the protein expression of TLR4(t=3.718,P=0.002)and NF-κB(t=5.892,P<0.001)and TNF-α(t=9.137,P<0.001)in renal tissue were significantly increased.After treatment with astragalosideⅣ,renal pathological damage was reduced,renal tissue TLR4(t=12.759,P<0.001),NF-κB(t=14.442,P<0.001)and TNF-α(t=7.156,P<0.001)mRNA expression was significantly down regulated,TLR4(t=3.782,P=0.001)and NF-κB(t=3.102,P=0.006)and TNF-α(t=3.023,P=0.007)protein expression decreased significantly.Conclusion AstragalosideⅣcan reduce the expression of inflammatory factors in ischemia-reperfusion rats,and its mechanism may be related to the inhibition of TLR4/NF-κB signal pathway.
作者
殷春晓
马亚琼
刘萍
陈庆云
YIN Chun-xiao;MA Ya-qiong;LIU Ping;CHEN Qing-yun(Department of Nephrology and Rheumatism,Nanyang Second People's Hospital,Nanyang 473000,China)
出处
《社区医学杂志》
CAS
2022年第8期430-434,共5页
Journal Of Community Medicine
