回回甘松饮含药血清对大鼠肾小球系膜细胞增殖、周期及FN，Col1α1，Col4α1基因表达的影响  

Effects of Huihui Gansong Yin on Proliferation of Rat Glomerular Mesangial Cells,Cell Cycle and Expression of FN,Col1α1 and Col4α1 mRNA

作　　者：袁玲[1,2] 南一[2] 吴宥熹 徐俊[1] 黄秀兰[1] 

机构地区：[1]中央民族大学中国少数民族传统医学研究院,北京100081 [2]宁夏医科大学回医药现代化省部共建教育部重点实验室,银川750004

出　　处：《中国实验方剂学杂志》2014年第4期135-140,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81202774,81102893)

摘　　要：目的:探讨回回甘松饮(HGY)含药血清对高糖诱导的大鼠肾小球系膜细胞(MCs)增殖、细胞周期、纤维连接蛋白(FN)、Ⅰ型胶原α1多肽链(Col1α1)、Ⅳ型胶原α1多肽链(Col4α1)基因表达的影响。方法:采用中药血清药理学方法制备含药血清,将大鼠随机分为空白对照组(给予蒸馏水10 mL·kg-1)、格列喹酮(10 mg·kg-1)组、HGY(10,5 g·kg-1)组,每组各10只,每日灌胃给药2次,连续3 d后取大鼠血清。选取HBZY-1细胞株进行实验,共分为5组:10%空白血清组、高糖+10%空白血清组、高糖+10%格列喹酮含药血清组(10 mg·kg-1)、高糖+10%HGY含药血清组(10,5 g·kg-1)。细胞以2×104/mL接种于96孔细胞培养板上,分别于药物处理后24,48,72 h时,采用CCK-8法检测MCs增殖情况;细胞以2×104/mL接种于25 cm2细胞培养瓶中,于药物干预48 h时收集细胞,分别采用流式细胞术检测细胞周期变化,real-time PCR方法检测FN、Col1α1及Col4α1 mRNA的表达。结果:与10%空白血清组比较,高糖(30 mmol·L-1)能够诱导MCs增殖(P<0.01),并能使G0/G1期细胞比例减少(P<0.01),S期比例增加(P<0.01),能够使MCs中FN,Col1α1,Col4α1 mRNA表达量明显升高(P<0.01)。在上述含药血清干预的48,72 h时,各含药血清组MCs均受到一定程度抑制,发生G1/S期阻滞,并可降低高糖条件下MCs中FN,Col1α1,Col4α1 mRNA表达量,与高糖+10%空白血清组比较,具有显著性差异(P<0.01或P<0.05)。结论:HGY含药血清可使大鼠MCs在高糖环境下发生G1/S期阻滞,抑制其增殖,其机制可能与调控FN,Col1α1及Col4α1 mRNA表达有关。Objective： To investigate the effects of Huihui Gansong Yin （HGY） on the proliferation of rat glomerular mesangial cells （MCs）, cell cycle, fibronectin （FN）, collagen type Ⅰ alpha 1 （Col1α1） and collagen typelV alpha 1 （Col4α1） induced by high glucose. Method： The serum containing HGY was prepared by adopting the method of serum pharmacology of TCM. Rats were divided into the blank group （distilled water,10 mL-kg-1）, glurenorm group （10 mg.kg-1）, HGY group （5, 10 g .kg-1）. There were 10 rats in each group. At the end of the 3th day serum was obtained. HBZY-1 cells （MCs） were divided into 5 groups： 10% blank serum group, high-glucose ＋ 10% blank serum group, high-glucose ＋ 10% serum containing glurenorm 10 mg .kg-1 , high-glucose ＋ 10% serum containing HGY 10, 5 g .kg-1 MCs were cultured in 96-well plates （2 × 10^4/mL）. After 24, 48, 72 h incubation with above indicated administration, the CCK-8 assay was used as a qualitative index of cell proliferation. Another MCs were cultured in 25 cm2 cell culture bottles （2 × 10^4/mL）. The cells were harvested 48h after treatment. Cell cycle analysis was performed by flow cytometry. FN, Col1α1 and Col4α1 mRNA were measured using real-time PCR. Result： Compared with 10% blank serum group, high- glucose （30 mmol .L-1） induced obvious proliferation of the MCs （P 〈0.01 ）. High-glucose induced decrease of cells in G0/G1 phase （P 〈0.01 ）, but increased cells in S phase （P 〈0.01 ）. High-glucose induced high levels of the expression of FN, Col1α1 and Col4α1 mRNA in MCs （P 〈0.01）. The above indicated administration, no matter for 48 h or 72 h, suppressed high glucose induced MCs proliferation and arrested with the cell cycle at G1/ S phase, meanwhile, attenuated the expression of FN, Col1α1 and Col4α1 mRNA in MCs induced by high glucose （P 〈0.01 or P 〈0.05）. Conclusion： The serum containing HGY could arrest with the cell cycle at G1/S phase and inhibit the proliferation of MCs

关 键 词：回回甘松饮 肾小球系膜细胞 细胞增殖 细胞周期 纤维连接蛋白 Ⅰ型胶原α1多肽链 Ⅳ型胶原 α1多肽链 

分 类 号：R285.5[医药卫生—中药学]