摘要
目的:观察中药复方制剂消可宁对高糖培养的大鼠肾小球系膜细胞增殖的抑制作用。方法:实验于2004-04/11在南京医科大学第一附属医院内分泌代谢研究中心细胞培养研究室完成。①中药复方制剂消可宁(由制大黄、制附子、生黄芪组成,南京军区南京总医院制剂科生产,药品批号:院临(2003)第01017号,规格为90mL/袋)。大鼠系膜细胞株HBZY-1购自武汉大学中国典型培养物保藏中心。②将购入的系膜细胞株消化培养,取培养4～8代的细胞,吸去培养瓶中的上清液,消化离心后混匀,锥虫蓝染色观察细胞活性。③不同刺激时间系膜细胞增殖实验设立3组,正常糖浓度组每孔加入5.6mmol/L葡萄糖溶液200μL,高糖对照组每孔加入30mmol/L葡萄糖溶液200μL,高糖+消可宁组每孔加入30mmol/L葡萄糖与0.06g/mL消可宁混合液200μL,6孔/组。各组分别于培养24,48,72h后向板孔中加5g/L的4-甲基偶氮四唑蓝20μL,于酶标仪492nm波长处检测吸光度值。④不同浓度消可宁刺激高糖环境下系膜细胞增殖实验设立7组,高糖对照组每孔加入30mmol/L葡萄糖溶液200μL,消可宁20,40,60,80,120,200g/L组每孔加入30mmol/L葡萄糖+对应浓度的消可宁混合液200μL,6孔/组。培养72h后各组向板孔中加5g/L的4-甲基偶氮四唑蓝20μL,同法检测吸光度值。结果:①系膜细胞活性观察:原代培养的4～8代系膜细胞,经锥虫蓝染色,着色的死亡细胞数<5%,未着色活细胞数>95%。②不同刺激时间各组系膜细胞增殖情况的比较:与正常糖浓度组比较,高糖对照组于高糖刺激24h即可促进系膜细胞的增殖,且这种作用在刺激48h时最为明显,至72h细胞数量开始减少,但仍明显高于正常糖浓度组(0.527±0.047,0.659±0.018,P<0.05)。与高糖对照组比较,高糖+消可宁组在24h内并未表现出抑制系膜细胞增殖的作用,而刺激48h后显示出抑制趋势,但差异无显著性意义,于72h后表现出较强的抑制系�
AIM： To observe the effect of compound Chinese medicine Xiaokening in suppressing the proliferation of mesangial cells in rats cultured with high glucose. METHODS： The experiment was conducted in Cultural Department of Cells, Center for Endocdnology and Metabolism, First Hospital Affiliated to Nanjing Medical University between Apdl and November 2004. （1） Xiaokening （made from Dahuang, Zhifuzi and Shenghuangqi, and was prepared by the Department of Pharmaceutic,s, General Hospital of Chinese PLA of Nanjing Military Area＇with the batch number of （2003） 01017, 90 mL/pack. （2） Intercapillary cell strains were cultured to obtain the cells from the 4^th to the 8^th passage, and the supematant in cultural bottles was gotten dd off, and samples were mixed after digestion and centdfugation. Trypan blue stain was performed to study the activity of cells. （3） Intercapillary cells stimulated at different times were divided into 3 groups with 6 pores in each group. Normal glucose group was added with 200 μL of glucose solution （5.6 mmol/L for each pore）, high-concentration glucose groupwas added with 200μL glucose solution （30 mmol/L for each pore）, and high-glucose and Xiaokening group was added with 200 p,L mixed solution （30 mmol/L glucose and 0.06 g/mL of Xiaokening for each pore）, Samples in 3 groups were added with 20 p,L of MTT （5 g/L） respectively at 24, 48 and 72 hours after culture. And the absorbance at 492 nm of meter tech. （4） There were 7 groups for proliferation test of intercapillary cells under high-glucose stimulated by different concentrations of Xiaokening. 30 mmol/L glucose solution was added into each pore of high-glucose control group, while 20, 40, 60, 80, 120 and 200 g/L Xiaokening groups were added with 200 μL mixed solution of 30 mmol/L glucose and Xiaokening of corresponding dose with 6 pores in each group. 72 hours after the culture, 20 μL of MTT （5g/L） was added into each group, and the absorbance was determined with the same method. RESULTS：
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第12期2371-2374,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
江苏省博士后基金特别资助课题(068NA0301)
江苏省人事厅六大人才高峰战略工程资助课题(2005A6)~~
