经穴注射BMSCs联合中药对糖尿病大鼠缺血后肢毛细血管密度的影响  

Acupoint injection of BMSCs combined with Chinese herbs for capillary density in ischemic hind limb of diabetes mellitus rats

作　　者：黄凤[1,2] 续畅 李彬[1] 王少松[1] 杜鑫[1] 国笑 荣培晶[2] HUANG Feng;XU Chang;LI Bin;WANG Shaosong;DU Xin;GUO Xiao;RONG Peijing(Beijing Hospital of TCM,Capital Medical Universit;Institution of Acupuncture and Moxibustion,China Academy of Chinese Medical Science;Beijing Shijitan Hospital,Capital Medical Universit)

机构地区：[1]首都医科大学附属北京中医医院,北京100010 [2]中国中医科学院针灸研究所,北京100700 [3]首都医科大学附属北京世纪坛医院

出　　处：《中国针灸》2018年第9期969-977,共9页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金青年项目：81503580; 北京市教育委员会科技发展计划一般项目：KM 201710025023; 北京市医院管理局“青苗”计划专项：QML 20171002; 针灸神经调控北京市重点实验室：BZ 0437

摘　　要：目的:观察经穴注射骨髓间充质干细胞(BMSCs)联合益气活血中药对糖尿病(DM)大鼠缺血后肢骨骼肌毛细血管密度及小动脉密度的影响。方法:80只SD大鼠按1∶7比例随机分为2组:正常假手术组10只,其余70只一次性腹腔注射链脲佐菌素(STZ,50.0 mg/kg)制成DM大鼠模型,再等比例随机分为7组,即DM假手术组、DM缺血组、中药组(益气活血中药灌胃)、局部注射组(局部注射BMSCs)、局部注射+中药组(局部注射BMSCs配合益气活血中药灌胃)、穴位注射组(穴位注射BMSCs)、穴位注射+中药组(穴位注射BMSCs配合益气活血中药灌胃),每组10只。局部注射组、局部注射+中药组取缺血组织与正常组织交界处等距离选取5个点为注射部位注射BMSCs,注射量及浓度同穴位注射组。穴位注射:穴取"三阴交""照海""环跳""后三里""阳陵泉",每只大鼠共注射100μL浓度为1×107/m L的BMSCs,每处注射20μL。灌胃中药为煎煮浓缩成含生药1.5 kg/L的益气活血中药(生黄芪120 g、党参120 g、炙甘草48 g、当归120 g、鸡血藤120 g、川牛膝48 g)。非中药组蒸馏水灌胃。应用免疫组织化学SP二步法检测大鼠骨骼肌中α-平滑肌肌动蛋白(α-actin)、血小板内皮细胞黏附分子CD31、血管性血友病因子(v WF)的表达。结果:干预结束后21 d,各组内术侧较健侧后肢α-actin、CD31(除中药组外)表达均有所升高(P<0.05,P<0.01);中药组、局部注射组、局部注射+中药组、穴位注射+中药组术侧与健侧比较,v WF表达均降低(P<0.05,P<0.01)。术侧组间比较,穴位注射+中药组与正常假手术组、DM假手术组、DM缺血组、局部注射组比较,骨骼肌中α-actin表达明显升高(P<0.05,P<0.01);穴位注射组、穴位注射+中药组、局部注射+中药组大鼠后肢骨骼肌CD31表达与正常假手术组、DM假手术组、DM缺血组比较明显升高(P<0.05,P<0.01);穴位注射+中药组较中药组和局部注射组CD31表达升高(均P<0.05);局�Objective To observe the effect of acupoint injection of bone mesenchymal stem cells（BMSCs） combined with Chinese herbs of benefiting qi for activating blood circulation for capillary density and arterioles density in skeletal muscle in ischemic hind limb of diabetes mellitus（DM） rats. Methods A total of 80 rats were randomized into a normal sham operation group（10 rats） and a model group（70 rats）. Disposable intraperitoneal injection of streptozotocin（STZ, 50.0 mg/kg） was used to establish DM model, and the rats in the model group were randomized into 7 subgroups, 10 rats in eachone. The subgroups were the DM sham operation group, DM ischemic group, Chinese herb group（intragastric herbs of benefiting qi for activating blood circulation）, local injection group（BMSCs local injection）, local injection ＋ Chinese herb group（BMSCs local injection combined with intragastric herbs of benefiting qi for activating blood circulation）, acupoint injection group（BMSCs acupoint injection）, acupoint injection ＋ Chinese herb group（BMSCs acupoint injection combined with intragastric herbs of benefiting qi for activating blood circulation）. The local injection was phosphate buffer（PBS） injection at the equidistant 5 points along the line between the ischemic tissue and the normal tissue a time. The acupoints were ＂Sanyinjiao＂（SP 6）, ＂Zhaohai＂（KI 6）, ＂Huantiao＂（GB 30）, ＂Housanli＂（ST 36） and ＂Yanglingquan＂（GB 34）. 100 μL BMSCs with 1×107/mL was totally injected at the above acupoints for one rat, 20 μL an acupoint. 1.5 kg/L Chinese herbs were applied by intragastric administration, including 120 g Radix Astragali, 120 g Codonopsis, 48 g Radix Glycyrrhiza, 120 g Angelica sinensis, 120 g Blood Rattan, 48 g Achyranthes bidentata. Intragastric distilled water was used in the other non-Chinese herb groups. The expressions of α-smooth muscle actin（α-actin）, latelet endothelial cell adhesion molecule（CD31） and von willebrand factor（v WF�

关 键 词：糖尿病 下肢缺血 α-平滑肌肌动蛋白(α-actin) 血小板内皮细胞黏附分子(CD31) 穴位注射 骨骼间充质干细胞 益气活血 毛细血管密度 

分 类 号：R245.95[医药卫生—针灸推拿学]