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1、非酒精性脂肪性肝病兔肾组织硫化氢与一氧化氮变化谭华炳 1，王一平 1，任永生 2，许 丽 1，胡小林 3，朱德文 3摘要 目的：观察非酒精性脂肪性肝病（NAFLD）兔肾组织硫化氢（H 2S） 、一氧化氮（NO）浓度，探讨 H2S、NO 在 NAFLD 肾损害中的作用。方法：40 只日本大耳白兔数字法随机分为重度 NAFLD 组（重度组）、轻度 NAFLD 组（轻度组） 、空白对照组（对照组）。重度组给予高脂饲料 160 g/(兔d)，轻度组给予高脂饲料 80 g/(兔d) + 普通饲料 80 g/(兔d)，对照组给予普通饲料 160 g/(兔d) 。均饲养 13 w。实验前后采集血浆标本，检测
2、甘油三酯（ TG）、胆固醇（ TC）。肾组织匀浆检测 NO、H 2S 浓度。免疫组织化学染色检测肾组织 -平滑肌肌动蛋白（-SMA）的表达。HE 染色，光镜观察肝组织病理学。结果： TC、 TG：饲养前重度组、轻度组、对照组 TC、TG 比较差异无统计学意义（P0.05），饲养后重度组 TC、TG 分别为(32.121.25)、(6.022.12) mmol/L，轻度组 TC、TG 分别为(18.342.10)、(4.391.93) mmol/L，均高于饲养前 (P0.01)，饲养后重度组 TC、TG 高于轻度组( P0.01)。 肾组织 NO：重度组(16.247.8mol/g 蛋白)、轻度
3、组(22.545.4mol/g蛋白)与对照组( 27.329.3mol/g蛋白)比较，有显著差异(P 0.01, P0.05)，重度组与轻度组比较有显著差异 (P0.05)。 肾组织 H2S 浓度：与对照组比较，重度组和轻度组肝组织 H2S 明显下降 (P0.01)，重度组低于轻度组，比较无显著差异(P0.05) 。 肾组织 -SMA 检测：对照组兔肾脏中的 -SMA 主要存在于血管壁；重度 NAFLD 组兔除血管壁外，肾小球系膜区及肾间质均有表达；轻度组较重度组 -SMA 表达减轻。 肝脏病理学：重度组肝脏病理学改变呈重度 NAFLD，轻度组呈轻度中度 NAFLD。结论：在 NAFLD 的发
4、生、发展过程中肾组织 H2S、NO 下降 ，NO、H 2S 可能参与 NAFLD 肾损害过程。关键词 非酒精性脂肪性肝病；肾组织；硫化氢；一氧化氮；-平滑肌肌动蛋白；动物Chang of Nitrogen Oxide and Hydrogen Sulfide Density in Rabbit Kidney Tissue With Nonalcoholic Fatty Liver Disease Tan Huabing, Wang Yiping, Xu Li, et al. Department of Infectious disease, Peoples Hospital of Hubei
5、medical university. Shiyan City 442000, ChinaAbstract Objective: To observate the changes of Nitric oxide(NO) and Hydrogen sulfide (H2S) in kidney tissue of the rabbit with Nonalcoholic fatty liver disease(NAFLD), discusses function of H2S and NO in injured kidney of NAFLD. Methods: 40 Japanese whit
6、e rabbits were randomly divided into severe group with NAFLD, mild group with NAFLD, blank control group. Severe group was given high fat diet 160g/rabbit.day, mild group were given high fat diet 80g /rabbit.day and normal diet 80g/rabbit.day, blank control group was given normal diet 160g/rabbit.da
7、y. The raising cycle is 13 weeks. Plasma samples collected before and after experiment, the samples in accor- dance with the equipment, reagents, experimental operation of routine testing before and after triglyceride(TG), cholesterol(TC). The Density of NO and H2S in kidney tissue were tested. Meth
8、ods Immunohistochemistry staining was used to detect the expression of -SMA in Rabbit Kidney Tissue. HE staining, Pathological changes of liver tissue were observed With the aid of Light microscope 基金项目 湖北省教育厅科技项目(D20102104)。十堰市科技局科技项目(2010st33)。湖北医药学院学生科研项目（2010XSA19） 。Hubei Province Education depa
9、rtment technological project (D20102104). Shiyan Science and technology Bureau technological project (2010st33). Hubei medicine university student scientific research item (2010XSA19).作者单位：湖北医药学院：1 附属人民医院；2 基础医学院；3 第三临床学院学生作者简介 谭华炳，男，主任医师，从事感染性疾病临床、科研、教学工作。通讯作者 王一平，男(1957-)，主任医师，从事代谢性肝病临床工作。Email: .
10、 电话：(0719)8637659.Results: TC and TG level: Before keeping severe group, mild group and blank control group there was no significant difference(P0.05), after the feeding severe group of TG and TC Respectively is (32.121.25)、(6.022.12)mmol/L, mild group of TG and TC Respectively is (18.342.10)、(4.391
11、.93) mmol/L, After the feeding, there was a significant difference compared in every group (P0.01). Severe group of TG and TC higher than mild group, there was a significant difference compared (P0.01). The level of NO in kidney tissue: Severe group of NO in kidney tissue is (16.247.8)mol/g.pro, mil
12、d group of NO in kidney tissue is (22.545.4)mol/g.pro, there was a significant difference compared with blank control group (P0.01, P0.05), there was a significant difference between severe group and mild group(P0.05). The level of H2S in kidney tissue: Severe group and mild group significantly decr
13、eased in liver tissue, there was a significant difference compared with blank control group(P0.01), there was no significant difference between Severe group and mild group (P0.05). Detected -SMA in kidney tissues: The blank control group Rabbit kidney of -SMA mainly existed in the blood vessel walls
14、, Mesangial area and renal stroma also expressed -SMA except the blood vessel walls in Severe NAFLD group, the expression of -SMA in mild group decreased compared with mild group. (5) Liver pathology: severe group of moderate was severe NAFLD, mild group was mild to moderate NAFLD, blank control gro
15、up was normal liver. Conclusions: H2S and NO participated the occurrence and development of NAFLD, it is the Future direction through interventing the NO and H2S possiblely. The levels of H2S and NO decrease in the procress of the occurrence and development of NAFLD, it is possible that the NO and H
16、2S participate in the procress of kidney damage. Keywords：NAFLD; Nephridial tissue; H 2S; NO; -SMA ; Animal测量肾内动脉阻力指数（RI）被广泛应用于评价肾内血管功能状态，甚至预测肾功能的变化。肝硬化患者 RI 数呈升高的趋势 1。非酒精性脂肪性肝病（nonalcoholic fatty liver disease, NAFLD）患者左RI 较健康成人降低 2，提示 NAFLD 可能存在肾脏受损。一氧化氮（Nitric oxide, NO）参与肝硬化和糖尿病肾病的发生、发展。硫化氢（ Hydrogen sulfide, H2S）参与肝硬化的发病过程 3, 4。NO 主要在肝实质细胞和 Kupffer 细胞中产生；肝脏是体内 H2S 生成的主要部位，对维持循环血液中 H2S 的
