甲基苯丙胺对大鼠肾脏损伤研究
发布时间：2018-08-11 15:22
【摘要】：《2015中国禁毒报告》显示[1],全国共破获毒品犯罪案件14.59万起,缴获各类毒品合计68.96吨,其中冰毒类毒品25.9吨,氯胺酮11.2吨,海洛因9.3吨、大麻4吨。全国累计登记吸毒人数295.5万名,其中滥用新型毒品人员达145.9万名,占吸毒人数的49.4%,首次超过海洛因滥用群体比例49.3%,且在新型毒品滥用者中,滥用甲基苯丙胺人员达119万,占新型毒品吸毒人数的81.6%,说明中国毒品滥用结构发生了深刻的变化;以甲基苯丙胺为主体的苯丙胺类兴奋剂(AST)滥用与依赖在我国和世界范围内快速蔓延,大有取代海洛因、可卡因等传统毒品的趋势,我国面临着继海洛因之后的甲基苯丙胺滥用的威胁。云南省地处我国西南,其边境地区环境优美,气候宜人,民族众多,边境贸易繁荣,自古就是历史文化风景名胜。自改革开放以来,从“金三角”毒源地经我国云南边境地区的毒品运输量愈来愈大,毒品运输愈加频繁,严重危害边境人民的正常生活和正常的经济秩序,也使得我国从过去毒品的过境国逐步变成制造、生产、销售和消费“一条龙”式的受害国。目前由于新型毒品尤其以甲基苯丙胺(冰毒)滥用人群较多,故本研究就甲基苯丙胺对肾脏损伤予以探讨。[目的]探讨滥用新型毒品甲基苯丙胺(methamphetamine,MA)对肾脏损伤机制的研究,本研究分为两部分,第一部分为研究甲基苯丙胺损伤人肾近曲小管上皮细胞(HK-2cells,HK-2细胞),采用MTS法进行细胞毒试验及细胞凋亡情况,以及急性损伤标志物中性粒细胞明胶酶相关脂质运载蛋白(Ngal)、肾损伤因子(Kim-1)的表达、分泌特点,并对其在甲基苯丙胺肾毒性损伤中的意义进行探讨。第二部分建立甲基苯丙胺对大鼠模型肾脏损伤模型,研究甲基苯丙胺对大鼠肾损伤机制。[方法]观察甲基苯丙胺对人肾近曲小管上皮细胞损伤作用。实验分组如下:正常对照组(仅加入培养液),对照组采用浓度分别为0.75mmol/L、1mmol/L、1.5mmol/L、2mmol/L、3mmol/L、4mmol/L、6mmol/L、8mmol/L、12mmol/L、16mmol/L甲基苯丙胺作用于体外培养人肾近曲小管上皮细胞,通过MTS法观察甲基苯丙胺对HK-2的影响,流式细胞术观察细胞周期改变和凋亡情况,ELISA法检测培养上清中Ngal与Kim-1的分泌情况。通过建立甲基苯丙胺急性、亚急性、慢性大鼠模型,取大鼠肾组织进行切片、HE染色及电镜扫描。[结果](1)甲基苯丙胺作用人肾小管上皮细胞24h,显微观察细胞形态病使用MTS法测定细胞生存率。甲基苯丙胺在0.75mmo1～16mmol/L浓度范围内,随着甲基苯丙胺浓度的增加,HK-2细胞的生存率相应降低。增殖实验结果显示:与相同时间内的对照组相比,浓度达6mmol/L～16mmol/L时甲基苯丙胺作用HK-2细胞24h差异有统计学意义(P0.05)。(2)甲基苯丙胺诱导HK-2细胞凋亡结果:0.75mmol/L 组:20.51%;1mmol/L 组:24%;1.5mmol/L 组:31.96%;2mmol/L 组:30.15%;3mmol/L 组:31.8%;4mmol/L 组:26.9%;6mmol/L 组:99%;8mmol/L 组:98%;12mmol/L 组:99.16%;16mmol/L 组:99.8%。(3)NGAL结果:8mmol/L组与对照组在相比,在12h内,随着作用时间延长,肾小管上皮细胞分泌NGAL量增加。12h后肾小管上皮细胞分泌NGAL逐渐减少。(4)KIM-1结果:8mmol/L组与对照组在相比,在48h内,随着作用时间延长,肾小管上皮细胞分泌KIM-1量增加。[结论](1)甲基苯丙胺对HK-2细胞的有抑制增殖作用,浓度达6mmol/L～16mmol/L时甲基苯丙胺作用HK-2细胞24h抑制作用越强。(2)甲基苯丙胺对HK-2细胞的促进凋亡。(3)在12h内,随着作用时间延长,肾小管上皮细胞分泌NGAL量增加。12h后肾小管上皮细胞分泌NGAL逐渐减少。(4)随着作用时间延长,肾小管上皮细胞分泌KIM-1量增加。(5)MA对肾脏有明显毒性作用。
[Abstract]:According to China's Drug Control Report in 2015, a total of 145,900 drug-related crimes were cracked and 68.96 tons of all kinds of drugs were seized nationwide, including 25.9 tons of methamphetamines, 11.2 tons of ketamine, 9.3 tons of heroin and 4 tons of cannabis. Among the new drug abusers, 1.19 million people abused methamphetamine, accounting for 81.6% of the new drug abusers, indicating that the structure of drug abuse in China has undergone profound changes; the abuse and dependence of amphetamine-type stimulants (AST), mainly methamphetamine, in China and around the world Yunnan Province is located in the southwest of China, with beautiful environment, pleasant climate, numerous nationalities and prosperous border trade. Since the reform and opening up, it has been a historical and cultural scenic spot. Drug traffic in the "Golden Triangle" is increasing in the border areas of Yunnan, China. Drug traffic is becoming more frequent, seriously endangering the normal life and normal economic order of the border people. It has also gradually transformed China from a drug transit country into a "one-stop" victim country of manufacturing, production, sales and consumption. Methamphetamine (MA) is a new drug, especially methamphetamine (methamphetamine) is abused in a large number of people, so this study on methamphetamine-induced renal damage to explore. [Objective] To explore the abuse of methamphetamine (MA) on the mechanism of renal damage, this study is divided into two parts, the first part is to study methamphetamine-induced renal damage in human proximal meander. Tubular epithelial cells (HK-2 cells, HK-2 cells) were used for cytotoxicity test and apoptosis by MTS method. The expression and secretion characteristics of neutrophil gelatinase-associated lipid carrier protein (Ngal) and kidney injury factor (Kim-1) in acute injury markers were studied. The significance of MTS in methamphetamine nephrotoxic injury was also discussed. [Methods] The effects of methamphetamine on renal proximal convoluted tubular epithelial cells in rats were observed. The experimental groups were as follows: normal control group (only adding culture medium), control group with concentrations of 0.75 mmol/L, 1.5 mmol/L, 2 mmol/L, respectively. L, 3mmol/L, 4mmol/L, 6mmol/L, 8mmol/L, 12mmol/L, 16mmol/L methamphetamine acted on cultured human renal proximal tubular epithelial cells in vitro. MTS method was used to observe the effect of methamphetamine on HK-2, flow cytometry was used to observe cell cycle changes and apoptosis. ELISA was used to detect the secretion of Ngal and Kim-1 in culture supernatant. [Results] (1) Methamphetamine treated human renal tubular epithelial cells for 24 hours. Cell survival rate was determined by MTS method. Methamphetamine concentration ranged from 0.75 mmo1 to 16 mmol/L, with the concentration of methamphetamine. The proliferative test showed that there was a significant difference in the effect of methamphetamine on HK-2 cells at the concentration of 6 mmol/L to 16 mmol/L for 24 hours (P 0.05). (2) Methamphetamine-induced apoptosis of HK-2 cells: 0.75 mmol/L group: 20.51%; 1 mmol/L group: 24%; 1.5 mmol/L group: 1.5 mmol/L; 2. L group: 31.96%; 2 mmol/L group: 30.15%; 3 mmol/L group: 31.8%; 4 mmol/L group: 26.9%; 6 mmol/L group: 99%; 8 mmol/L group: 98%; 12 mmol/L group: 99.16%; 16 mmol/L group: 99.8%. (3) NGAL results: 8 mmol/L group compared with the control group, in 12 hours, the amount of NGAL secreted by renal tubular epithelial cells increased. (4) KIM-1 gradually decreased. (4) KIM-1 secretion of renal tubular epithelial cells increased with the prolongation of the treatment time in the 8mmol/L group compared with the control group. [Conclusion] (1) Methamphetamine inhibited the proliferation of HK-2 cells. When the concentration reached 6mmol/L to 16mmol/L, the inhibition of methamphetamine on HK-2 cells became stronger in 24h. (2) Methamphetamine inhibited the proliferation of HK-2 cells. Amine promoted apoptosis of HK-2 cells. (3) Within 12 hours, NGAL secreted by renal tubular epithelial cells increased with the prolongation of treatment time. NGAL secreted by renal tubular epithelial cells decreased gradually after 12 hours. (4) KIM-1 secreted by renal tubular epithelial cells increased with the prolongation of treatment time. (5) MA had a significant toxic effect on kidney.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.64
本文编号：2177392
[Abstract]:According to China's Drug Control Report in 2015, a total of 145,900 drug-related crimes were cracked and 68.96 tons of all kinds of drugs were seized nationwide, including 25.9 tons of methamphetamines, 11.2 tons of ketamine, 9.3 tons of heroin and 4 tons of cannabis. Among the new drug abusers, 1.19 million people abused methamphetamine, accounting for 81.6% of the new drug abusers, indicating that the structure of drug abuse in China has undergone profound changes; the abuse and dependence of amphetamine-type stimulants (AST), mainly methamphetamine, in China and around the world Yunnan Province is located in the southwest of China, with beautiful environment, pleasant climate, numerous nationalities and prosperous border trade. Since the reform and opening up, it has been a historical and cultural scenic spot. Drug traffic in the "Golden Triangle" is increasing in the border areas of Yunnan, China. Drug traffic is becoming more frequent, seriously endangering the normal life and normal economic order of the border people. It has also gradually transformed China from a drug transit country into a "one-stop" victim country of manufacturing, production, sales and consumption. Methamphetamine (MA) is a new drug, especially methamphetamine (methamphetamine) is abused in a large number of people, so this study on methamphetamine-induced renal damage to explore. [Objective] To explore the abuse of methamphetamine (MA) on the mechanism of renal damage, this study is divided into two parts, the first part is to study methamphetamine-induced renal damage in human proximal meander. Tubular epithelial cells (HK-2 cells, HK-2 cells) were used for cytotoxicity test and apoptosis by MTS method. The expression and secretion characteristics of neutrophil gelatinase-associated lipid carrier protein (Ngal) and kidney injury factor (Kim-1) in acute injury markers were studied. The significance of MTS in methamphetamine nephrotoxic injury was also discussed. [Methods] The effects of methamphetamine on renal proximal convoluted tubular epithelial cells in rats were observed. The experimental groups were as follows: normal control group (only adding culture medium), control group with concentrations of 0.75 mmol/L, 1.5 mmol/L, 2 mmol/L, respectively. L, 3mmol/L, 4mmol/L, 6mmol/L, 8mmol/L, 12mmol/L, 16mmol/L methamphetamine acted on cultured human renal proximal tubular epithelial cells in vitro. MTS method was used to observe the effect of methamphetamine on HK-2, flow cytometry was used to observe cell cycle changes and apoptosis. ELISA was used to detect the secretion of Ngal and Kim-1 in culture supernatant. [Results] (1) Methamphetamine treated human renal tubular epithelial cells for 24 hours. Cell survival rate was determined by MTS method. Methamphetamine concentration ranged from 0.75 mmo1 to 16 mmol/L, with the concentration of methamphetamine. The proliferative test showed that there was a significant difference in the effect of methamphetamine on HK-2 cells at the concentration of 6 mmol/L to 16 mmol/L for 24 hours (P 0.05). (2) Methamphetamine-induced apoptosis of HK-2 cells: 0.75 mmol/L group: 20.51%; 1 mmol/L group: 24%; 1.5 mmol/L group: 1.5 mmol/L; 2. L group: 31.96%; 2 mmol/L group: 30.15%; 3 mmol/L group: 31.8%; 4 mmol/L group: 26.9%; 6 mmol/L group: 99%; 8 mmol/L group: 98%; 12 mmol/L group: 99.16%; 16 mmol/L group: 99.8%. (3) NGAL results: 8 mmol/L group compared with the control group, in 12 hours, the amount of NGAL secreted by renal tubular epithelial cells increased. (4) KIM-1 gradually decreased. (4) KIM-1 secretion of renal tubular epithelial cells increased with the prolongation of the treatment time in the 8mmol/L group compared with the control group. [Conclusion] (1) Methamphetamine inhibited the proliferation of HK-2 cells. When the concentration reached 6mmol/L to 16mmol/L, the inhibition of methamphetamine on HK-2 cells became stronger in 24h. (2) Methamphetamine inhibited the proliferation of HK-2 cells. Amine promoted apoptosis of HK-2 cells. (3) Within 12 hours, NGAL secreted by renal tubular epithelial cells increased with the prolongation of treatment time. NGAL secreted by renal tubular epithelial cells decreased gradually after 12 hours. (4) KIM-1 secreted by renal tubular epithelial cells increased with the prolongation of treatment time. (5) MA had a significant toxic effect on kidney.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.64
【参考文献】
相关期刊论文 前10条
1 郭鑫;王大伟;;云南边境地区入境运输毒品管控对策研究[J];广西警察学院学报;2017年01期
2 何莉;王孟龙;;肝移植术后急性肾损伤的研究进展[J];北京医学;2015年09期
3 孔大为;刘志远;;《2015中国禁毒报告》[J];人民公安;2015年12期
4 高艺航;印璞;于庭;;中性粒细胞明胶酶相关脂质运载蛋白—新型急性肾损伤的早期诊断标志物[J];中国实验诊断学;2015年03期
5 李磊;;云南中缅边境毒品犯罪现状及打击对策[J];法制博览;2015年04期
6 刘雪梅;刘孟春;马瑞霞;刘丽秋;徐岩;;恶性肿瘤伴发急性肾损伤患者的临床特点分析[J];中华肾脏病杂志;2014年11期
7 孙妍蓓;刘必成;邹芸;潘家荣;陶源;杨敏;;1371例急性心肌梗死后急性肾损伤患者的临床分析[J];中华肾脏病杂志;2014年10期
8 王志刚;;云南边境地区毒品犯罪特点及对策研究[J];现代商贸工业;2008年02期
9 孙骅;刘志红;王生余;秦卫松;陈朝红;黎磊石;;马兜铃酸诱导肾小管上皮细胞损伤中Ngal和Kim-1的变化及其意义[J];肾脏病与透析肾移植杂志;2007年04期
10 陈立成,任希全,刘晓温;冰毒及其危害[J];中国医师杂志;2003年04期
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