不同提取方法对土鳖虫的生物活性物质的影响  

Effects of different extraction methods on promoting rat femoral fracture healing by external use of Tubiechong

作　　者：邵炳豪 张泽峰 王晶 邓玉莹 王春梅 SHAO Binghao;ZHANG Zefeng;WANG Jing;DENG Yuying;WANG Chunmei(School of Life Science,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学生命科学学院,北京100029

出　　处：《生物加工过程》2025年第2期168-173,共6页Chinese Journal of Bioprocess Engineering

基　　金：北京中医药大学科研发展基金(202212)。

摘　　要：土鳖虫是治疗骨折愈合的常用中药,常以乙醇长期浸泡的方式制备成药酒外用,但制备周期过长。本研究以骨折大鼠作为动物模型评价不同提取方法所得土鳖虫活性物质外用促进骨折愈合的效果,从而确定较优的提取方法。土鳖虫药材以50%乙醇分别在25℃(传统冷浸组)、60℃浸提(热浸组)或者50℃匀浆并浸提12 h(匀浆热浸组)后冻干。以改良的Einhorn法构建大鼠股骨骨折模型,记录处理后各组大鼠骨折愈合时间,在用药4周时取血清,酶联免疫吸附方法检测骨碱性磷酸酶、骨钙素以及Ⅰ型胶原C末端肽的含量,从而评价骨折愈合程度。结果发现:土鳖虫贴敷处理对大鼠外观、体质量、摄食及行为习惯无明显影响;土鳖虫提取物各组均能缩短骨折愈合时间,空白组大鼠愈合时间为28 d,匀浆热浸中剂量组愈合时间为21 d;匀浆热浸组和热浸组血清骨碱性磷酸酶、骨钙素含量、血清中Ⅰ型胶原C末端肽的含量均显著升高(P<0.01),匀浆热浸组效果更明显。因此,土鳖虫提取液外敷能够促进大鼠骨折愈合,匀浆热浸法制备的提取物活性最佳。Tubiechong,a commonly used traditional Chinese medicine for external treatment of fracture healing,is often prepared as medicinal liquor through long-term immersion in ethanol.In this study,three kinds of Tubiechong extracts obtained by different methods were evaluated with regards to their promotion of fracture healing using rats as animal model.The traditional Chinese medicine,Eupolyphaga sinensis Walker,was treated in 50%ethanol at 25℃(traditional cold soak group)and 60℃(hot soak group),respectively.The other one(homogenization hot soak group)was obtained after the insects were crushed,passed40 mesh sieve and then homogenized at 50℃for 12 h.The supernatants from these three groups were centrifuged and lyophilized separately for the following studies.The rat femoral fracture model was constructed according to the modified Einhorn method.The rats were divided into 10 groups(with 5 animals in each group):the control group of normal rats without fracture;the positive drug group;the blank solvent group;the traditional cold soak group;three hot soak groups(with low,medium and high dose);and three homogenization hot soak groups(with low,medium and high dose).Upon treatment,the fracture healing time was observed and recorded,and the serum was taken 4 weeks there after.The degree of fracture healing was evaluated by the contents of bone alkaline phosphatase,osteocalcin and C-terminal peptide of typeⅠcollagen measured by enzyme linked immunosorbent assay.As a result,no significant difference was observed on the appearance,weight change,feeding and behavior habits of the rats in all groups.The healing time of fractures could be shortened in all groups treated with Tubiechong extract.Compared with the rats in the blank solvent group with healing time of 28 days,the medium dose homogenization hot soak group showed shorter healing time(21 days).The serum bone alkaline phosphatase and osteocalcin contents were significantly higher in all three homogenization groups and the high dose hot soak group(p<0.01),while the c

关 键 词：土鳖虫 外敷 大鼠股骨骨折模型 热浸法 匀浆热浸法 

分 类 号：R943[医药卫生—药剂学] Q966[医药卫生—药学]