中药制剂承载丸对甲基强的松龙预处理的成骨细胞L型钙通道电流的影响  

Effect of Chinese herb Chengzai Wan on the calcium channel currents of osteoblasts pretreated with methylprednisolone

作　　者：陈燕平[1] 贾丙申[1] 黄克勤[2] 张文杰[3] 薛延[4] 周重光[1] 张万强[1] 

机构地区：[1]中国中医科学院望京医院药理研究室,北京市100102 [2]北京皇城股骨头坏死专科医院,北京市100220 [3]吉林大学基础医学院,吉林省长春市130021 [4]北京积水潭医院骨伤科研究所,北京市100035

出　　处：《中国组织工程研究与临床康复》2007年第23期4546-4549,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然基金项目(30472173)~~

摘　　要：目的:观察承载丸含药血清对甲基强的松龙预处理的MC3T3-E1成骨细胞L-钙通道电流的影响。方法:实验于2006-04/06在中国中医科学院望京医院药理实验室及吉林大学基础医学院生理教研室完成。①实验材料:MC3T3-E1(购于北京协和细胞中心);8个月龄SD大鼠10只,雌雄各半;中药制剂承载丸(由鹿角霜、肉苁蓉、续断、黄芪、当归、炙水蛭、杜仲、蛋衣、皂角刺、香附、乌药等21味中药组成);甲基强的松龙(批号H20040338,Pharmacia NV/SA生产)。②实验干预:成骨细胞(MC3T3-E1)传代培养。承载丸药粉60g,加水至200mL搅匀,取大鼠6只(雌雄各半),每只灌服2.5mL/d,灌服4d后动脉取血,制备含药血清。另取大鼠4只,同法灌服生理盐水,制成不含药血清。甲基强的松龙的浓度为1×10-5mol/L。③实验分组:将培养后的成骨细胞株分为4组:正常组(加入正常大鼠不含药血清)、模型组(加入正常大鼠不含药血清及甲基强的松龙)、承载丸低剂量组(加入甲基强的松龙及0.5mL承载丸含药血清)和承载丸高剂量组(加入甲基强的松龙及0.1mL承载丸含药血清)。④实验评估:24h后,用全细胞膜片钳技术记录每组10个细胞的L-钙通道电流,并测定其峰值电流。结果:①各组电流峰值测定结果:模型组峰值电流比正常组下降19.5%[(0.1839±0.0179),(0.2284±0.0209)nA,P<0.01];承载丸低剂量组和高剂量组比模型组分别升高37.4%和45.2%[(0.2526±0.0093),(0.2671±0.0120),(0.1839±0.0179)nA,P<0.01];承载丸高低剂量组相比差异有显著性意义(P<0.05)。②MG3T3E1细胞钙电流的特性分析:正常组,钙通道大约在-20mV时开放,最大峰值电流在+20～+30mV,反转电位在+60～+70mV。当在浴液中加入Co2+,可阻断此电流的大部分电流成份,其峰值电流为(0.1050±0.0097)nA,而钙离子激活剂Bayk8644增加了此电流(0.3335±0.0323)nA。结论:①甲基强的松龙抑制成骨细胞L-钙通道电流。②承载丸含药血清可�AIM： To observe the effect of Chengzai Wan on the current of L-type voltage-sensitive calcium channels （L-VSCCsC） of MC3T3-E1 osteoblasts pretreated with methylprednisolone. METHODS： The experiment was carried out at Laboratory of Pharmacology, Wangjing Hospital of China Academy of Traditional Chinese Medicine and Department of Physiology of School of Basic Medical Sciences, Jilin University from April to June 2006. MC3T3-E1 osteoblasts purchased from Cell Center of Peking Union Medical College were passaged. 60 g powder of Chengzai Wan （composed of cervi degelatinatum, cistanches, teasel root, astragalus mongholicus, angelica sinensis, roast leech, eucommia bark, egg membrane, spina gleditsiae, rhizoma cyperi, and combined spicebush root） was mixed with water until 200 mL, and orally given to six of ten eight-month-old SD rats （haft male, half female） 2.5 mL daily. Four days later, the blood from abdominal aorta was harvested to prepare the serum containing herbs. Other 4 rats were treated with the same volume normal saline to prepare the serum with no herb. The concentratiorl of methylprednisolone （No. 20040338, produced by Pharmacia NV-SA） was adjusted to 1×10^-5mol/L. The cultured osteoblasts were divided into 4 groups： normal group, which was given herb free serum, model group, which was given herb free serum and methylprednisolone, low dose group, which was added methylprednisolone and 0.5 mL serum containing herb, and high dose group, which was added methylprednisolone and 0.1 mL serum containing herb. Twenty-four hours later, the whole-cell patch clamp recording technique was employed to record L-VSCCsC of 10 cells in each group and determine the peak current. RESULTS：①The peak current of the model group was decreased by 19.5% compared with normal group [（0.183 9± 0.017 9）, （0.228 4±0.020 9） nA, P 〈 0.01]; that of the low dose and high dose groups was increased 37.4% and 45.2% compared with model group [（0.252 6±0.009 3）, （0.267 1±0.012 0）, （0.183 9±0.01

关 键 词：膜片钳术 钙通道 L型 甲泼尼松 成骨细胞 

分 类 号：R285.1[医药卫生—中药学]