摘要
目的研究盐酸吡格列酮对糖尿病大鼠肾组织Toll样受体4(TLR4)表达的影响。方法 57只雄性SD大鼠,分为实验组(n=48)和正常对照组(n=9)。实验组大鼠腹腔注射链脲佐菌素(STZ)50 mg/kg建立糖尿病大鼠模型,正常对照组注射等剂量无菌枸橼酸钠缓冲液。建模成功的45只大鼠随机分为5组,每组9只。吡格列酮5 mg/(kg·d)、吡格列酮10 mg/(kg·d)组、吡格列酮5 mg/(kg·d)联合TLR4特异性抑制剂Eritoran 5 mg/(kg·d)、吡格列酮10 mg/(kg·d)联合TLR4特异性抑制剂Eritoran 5 mg/(kg·d),未干预组。吡格列酮采用灌胃的方法给药,对照组给予等量生理盐水灌胃。联合应用TLR4特异性抑制剂Eritoran组于第5周给予腹腔注射Eritoran 5 mg/(kg·d),连续6周。于第8周末各组大鼠留24 h尿液测量尿微量白蛋白,采用Western blot法检测肾组织中TLR4表达水平。心尖部取血2 mL,测空腹血糖、血CRP。免疫组织化学染色检测各组大鼠肾组织中TLR4及过氧化物酶体增殖物激活受体γ(PPARγ)的表达。结果与对照组相比,实验组大鼠24 h尿微量蛋白、血CRP含量显著增高;肾组织TLR4的表达明显增加,治疗前与治疗后比较存在差异(P<0.05);与未干预组相比,各干预组大鼠24 h尿微量白蛋白、血CRP含量下降,肾组织TLR4的表达亦减弱,差异有统计学意义(P<0.05);10 mg/(kg·d)吡格列酮组与5 mg/(kg·d)吡格列酮组比较,肾组织TLR4的表达减弱,差异有统计学意义(P<0.05);10 mg/(kg·d)吡格列酮联合5 mg/(kg·d)Eritoran组与5 mg/(kg·d)吡格列酮联合5 mg/(kg·d)Eritoran组比较,肾组织TLR4的表达均明显减弱,但差异无统计学意义(P>0.05)。结论吡格列酮下调糖尿病大鼠肾脏组织TLR4表达,调节促炎和抗炎之间平衡,发挥抗炎作用。
Objective To investigate the effect of pioglitazone on the expression of Toll-like receptor 4 （TLR4） in renal tissue of diabetic rats. Methods The male Sprague Dawlry rats （ n = 57 ） were randomly divided into the control group （ n = 9） and experimental group （ n = 48）. The experimental rats were injected introperatoneally with 50 mg/kg streptozotocin （STZ） and the controls were given sodium citrate buffer instead. Then the experimental rats were randomized into five groups： pioglitazone 5 mg/（ kg ~ d）, pioglitazone 10 mg/（ kg ·d）, pioglitazone 5 mg/（ kg · d） plus TLR4-specific antagonist Eritoran 5 mg/（ kg · d）, pioglitazone l0 mg/（ kg · d） plus TLR4-specific antagonist Eritoran 5 mg/（ kg · d）, non-intervented group, with 5 rats in each group. The pioglitazone was administrated intragastrically, and normal saline was given to the contro group in the same way. Five weeks after pioglitazone administration, Eritoran was injected intraperitoneally for consecutive one week. At the end of the eighth week, 24-hour microalbuminuria and the serum concentrations of C-reactive protein （CRP） were determined by radioimmunoassay. Expressions of TLR4 and peroxisome proliferator-activated receptor y （PPARy） in kidney were determined by Western blotting and immunohistochemistry. Results Compared with the control group, the 24-hour urine protein, plasma concentration of CRP, the expression of TLR4 in kidney increased significantly in the experimental groups （P 〈 0.05）. Furthermore, these indicators in all intervention groups obviously decreased compared with the non-intervention group （ P 〈 0.05 ）. In addition, the expression of TLR4 in high-dose pioglitazone significantly decreased compared with low-dose pioglitazone （ m 〈 0.05 ）. The study also found that high-dose pioglitazone and TLR4 antagonist Eritoran could reduce the expression of TLR4 in the diabetic rats, but the difference from the group of low-dose pioglitazone plus Eritoran was not significant statistically （ P 〉 0.05）. Conclusion Pioglitazone may exert the anti-inflammatory action by decreasing the expression of TLR4 in renal tissue and regulating the balance between proinflammatory and anti-inflammatory.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2014年第8期793-797,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(30770155)
湖南省自然科学基金(12JJ3105)
