组蛋白去乙酰化酶抑制剂减轻草酸钙结晶肾损伤的实验研究

摘要

目的：观察组蛋白去乙酰化酶抑制剂辛二酰苯胺异羟肟酸（suberoylanilide hydroxamic acid，SAHA）对草酸钙结晶模型小鼠肾结晶沉积的作用，初步探讨 SAHA 对减轻草酸钙结晶肾损伤的机制。方法：24只 ICR 雄性小鼠随机分为4组，即空白对照组（A 组）和实验组（B、C、D 组）；B 组：50 mg/ kg 的生理盐水（NS）+100 mg/ kg 乙醛酸盐，C 组：50 mg/ kg 的DMSO +100 mg/ kg 乙醛酸盐，D 组：50 mg/ kg 的 SAHA +100 mg/ kg 乙醛酸盐；实验组给予乙醛酸盐6 h 前分别予以 NS、DMSO、SAHA 50 mg/ kg 等量腹腔注射，7 d 后处死全部小鼠。检测各组小鼠肾组织钙含量水平，丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽还原酶（GSH）的含量或活力，并且在光镜下观察各组小鼠肾组织切片中草酸钙结晶的分布，免疫组化及其半定量分析比较骨桥蛋白（OPN）、CD44分子在肾脏中的表达情况。结果：与 B、C 组相比，D 组小鼠的肾组织钙含量和MDA 含量显著下降（P 均<0．05），肾组织切片冯库萨染色显示草酸钙结晶沉积明显减少（P <0．05），OPN 与 CD44表达水平也明显下降（P <0．05）。而 B 组与 C 组肾结晶形成的差异无统计学意义。结论：研究结果首次证实 SAHA 对减少小鼠草酸钙肾结晶形成有预防作用，而且其机制可能与抑制氧化应激和降低 OPN、CD44表达水平有关。%Objective:To observe the effect of Histone Deacetylase inhibitor SAHA on kidney crystal formation in calcium oxalate Model Mice,and to investigate the underlying mechanism of renoprotective effect of SAHA. Methods:Twenty - four male ICR rats were randomly divided into four groups:namely blank control group(A group)and experiment groups( B、C、D group),B group:50 mg/ kg normal saline(NS)+ 100 mg/ kg glyoxylate,C group:50 mg/ kg DMSO + 100 mg/ kg glyoxylate,D group:50 mg/ kg SAHA + 100 mg/ kg glyoxylate. The rats in experiment groups were injected with 50 mg/ kg NS,DMSO,SAHA into abdominal cavity respectively 6 hours before glyoxylate. Rats were sacrified after seven days and the kidneys were harvested to detect calcium concentra-tion levels,malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione reductase(GSH). Furthermore,we observed the distribution of crystallization of calcium oxalate in kidney tissue by microscopy. Immunohistochemical staining and semi - quantita-tive analysis were used to detect the expression of osteopontin(OPN)and its cell surface receptor CD44 . Results:Compared with B、C group,the levels of the calcium concentration,MDA siginificantly decreased and crystallization of calcium oxalate in kidney tissue as well as the expression of OPN,CD44 were also lower in D group(all P < 0. 05). However,the crystallization of calcium oxalate be-tween B and C groups had no statistical differences. Conclusion:The results of this study firstly prove the preventive effect of SAHA treatment for kidney crystal formation and the mechanism may involves inhibition of oxidative stress and downregulating levels of OPN, CD44 .