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作 者:王慧 杨继红[1] 杜俊杰 WANG Hui;YANG Ji-hong;DU Jun-jie(Department of Nephrology,Beijing Hospital,National Center for Geriatric Medicine,Beijing 100730,China)
机构地区:[1]北京医院肾内科国家老年医学中心,北京100730
出 处:《中华实用诊断与治疗杂志》2022年第1期27-29,共3页Journal of Chinese Practical Diagnosis and Therapy
基 金:国家自然科学基金(81373803)。
摘 要:目的观察雷公藤甲素(triptolide, Tp)干预后脂多糖诱导的小鼠肾脏足细胞上皮-间质转化(epithelial-mesenchymal transition, EMT)相关蛋白表达变化,探讨Tp对小鼠肾脏足细胞EMT的抑制作用。方法采用CCK-8法检测小鼠肾脏足细胞活性,确定脂多糖浓度为5μg/L,Tp浓度为10μg/L。将小鼠肾脏足细胞分为对照组(不作处理)、脂多糖组(5 mg/L脂多糖溶液)和Tp+脂多糖组(10μg/L Tp+5 mg/L脂多糖溶液),处理24 h,采用Western blot法检测Toll样受体(Toll-like receptor, TLR)4、基质金属蛋白酶(matrix metalloproteinase, MMP)-9、紧密连接蛋白-1(zonula occluden-1, ZO-1)蛋白相对表达量。结果脂多糖组TLR4(1.24±0.04)、MMP-9(0.87±0.03)蛋白相对表达量均高于对照组(1.07±0.03、0.61±0.16)和Tp+脂多糖组(0.98±0.11、0.60±0.01)(P<0.05),ZO-1蛋白相对表达量(0.59±0.10)均低于对照组(0.88±0.14)和Tp+脂多糖组(0.82±0.14)(P<0.05);Tp+脂多糖组TLR4、MMP-9、ZO-1蛋白相对表达量与对照组比较差异均无统计学意义(P>0.05)。结论 Tp可下调小鼠肾脏足细胞TLR4、MMP-9表达,上调ZO-1表达,抑制脂多糖诱导的EMT。Objective To observe the change of mouse podocyte epithelial-mesenchymal transition(EMT) related proteins induced by lipopolysaccharide(LPS) after triptolide(Tp) treatment, and to investigate the role of Tp in inhibiting podocyte EMT. Methods The activity of mouse podocytes was detected by CCK-8, defining 5 mg/L and 10 μg/L as the concentrations of LPS and Tp, respectively. The mouse podocytes were divided into control group(with no treatment), LPS group(treated by 5 mg/L LPS solution), and TP+LPS group(treated by 10 μg/L Tp and 5 mg/L LPS), for 24 h. Western blot was used to detect the relative expressions of Toll-like receptor 4(TLR4), matrix metalloproteinase-9(MMP-9) and zonula occluden-1(ZO-1) proteins. Results The relative expressions of TLR4 and MMP-9 were higher in LPS group(1.24±0.04, 0.87±0.03) than those in control group(1.07±0.03, 0.61±0.16) and Tp+LPS group(0.98±0.11, 0.60±0.01)(P<0.05), the relative expression of ZO-1 was lower in LPS group(0.59±0.10) than that in control group(0.88±0.09) and Tp+LPS group(0.82±0.14)(P<0.05), and there were no significant differences in the relative expressions of TLR4,MMP-9 and ZO-1 between control group and Tp+LPS group(P>0.05). Conclusion Tp can down-regulate TLR4 and MMP-9, up-regulate ZO-1, and inhibit LPS induced EMT in mice.
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