平目中药全方及拆方含药血清对Graves眼病眼眶前脂肪细胞增殖和凋亡的影响  

Influences medicated serum of Pingmu Fang and its disassembled formulas on proliferation and apoptosis of orbital preadipocytes of Graves' ophthalmopathy

作　　者：张亚利[1] 王亚琼[2] 李红[2] 

机构地区：[1]上海中医药大学附属龙华医院脾胃病研究所上海200032 [2]上海中医药大学附属龙华医院内分泌科

出　　处：《北京中医药大学学报》2014年第3期184-189,I0004,共7页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家中医临床研究基地龙医团队项目(No.LYTD-10);国家自然科学基金项目(No.81072793,81373617);上海市教委科研创新重点资助项目(No.11ZZ114)

摘　　要：目的研究平目中药全方及拆方含药血清对Graves眼病眼眶前脂肪细胞增殖和成熟脂肪细胞凋亡的影响。方法体外培养Graves眼病患者眼眶前脂肪细胞,鉴定并传代。诱导前脂肪细胞分化,油红O染色鉴定成熟脂肪细胞。将平目中药全方拆方益气组、温阳组、化痰祛瘀组、益气温阳组、益气化痰祛瘀组、温阳化痰祛瘀组,制备全方组及各拆方组含药血清。MTT法观察平目中药全方及拆方对前脂肪细胞增殖活性的影响;流式细胞术测定平目中药全方及拆方对成熟脂肪细胞凋亡率的影响。结果原代培养细胞呈梭形,间接免疫荧光检测鉴定培养细胞为前脂肪细胞,细胞可分化为成熟脂肪细胞。MTT结果显示,平目中药全方及各拆方组均可降低前脂肪细胞增殖活性,其中全方组最为显著,益气温阳组其次,地塞米松组与空白对照组相比差异无统计学意义。全方组、化痰祛瘀组、益气温阳组、益气化痰祛瘀组、温阳化痰祛瘀组作用12 d后OD值出现明显差异(P<0.05),作用16 d后更为显著(P<0.01),地塞米松组差异无统计学意义。流式细胞术结果表明,与空白对照组相比,全方组、益气温阳组、益气化痰祛瘀组、温阳化痰祛瘀组的细胞凋亡率均升高(P<0.05),其中全方组凋亡率最高(10.5%±1.45%),益气温阳组其次(9.9%±0.60%),地塞米松组差异无统计学意义。结论平目中药全方及拆方对非活动期Graves眼病眼眶脂肪细胞积聚有不同的作用,全方组有整体优势,拆方组中以益气温阳组作用较明显,可能通过抑制前脂肪细胞增殖,促进脂肪细胞凋亡,减少眼眶脂肪细胞积聚,发挥治疗Graves眼病的作用。Objective To study the influences of medicated serum of Pingmu Fang and its disassembled formulas on proliferation and apoptosis of orbital preadipocytes of Graves＇ ophthalmopathy. Methods The orbital preadipocytes from the patients were cultured in vitro, identified, transferred and induced to differentiation, and then identified as mature adipocyte by using oil red O staining. The medicated serum samples were prepared and divided into whole Pingmu Fang group （whole formula group）, replenishing-qi group （disassembled formula group 1 ）, warming-yang group （disassembled formula group 2 ）, resolving-phlegm dispelling-stasis group （disassembled formula group 3 ） , replenishing-qi warming-yang group （disassembled formula group 4 ）, replenishing-qi resolving-phlegm dispelling-stasis group （ disassembled formula group 5 ） and warming-yang resolving-phlegm dispelling-stasis group （disassembled formula group 6 ）. The influences of medicated serum of Pingmu Fang and its disassembled formulas on the proliferation and apoptosis rate of orbital preadipocytes were observed respectively by using MTI＂ and flow cytometry. Results The results of MTF showed that the proliferation of orbital preadipocytes was inhibited in whole formula group and all disassembled formula groups, which was the most significant in whole formula group and followed by disassembled formula group 4, and there was statistical difference between dexamethasone group and blank group. After 12 d, OD had significant difference （P 〈 0.05 ） in whole formula group, disassembled formula group 3, disassembled formula group 4, disassembled formula group 5 and disassembled formula group 6, which was more significant after 16 d and had no statistical significance in dexamethasone group. The results of flow cytometry showed that compared with blank group, the apoptosis rate of orbital preadipocytes increased in whole formula group, disassembled formula group 4, disassembled formula group 5 and disassembled formula group 6 （P 〈 0

关 键 词：平目方 GRAVES眼病 非活动期 眼眶脂肪细胞 增殖 凋亡 大鼠 

分 类 号：R289.58[医药卫生—方剂学]