以Lipid A为靶点拮抗内毒素中药的筛选

目的利用生物传感器跟踪技术从抗炎中药中筛选出具有拮抗内毒素的中药,为进一步对中药拮抗内毒素活性成分的分离纯化提供实验依据。方法将革兰阴性细菌的脂质A(Lipid A)包被于生物传感器的非衍生板建立靶点,跟踪测定赤芍、大黄、黄芩等78种中药的水提物和醇提物与Lipid A的结合活性,再将筛选出的中药去鞣质后与定量的内毒素(LPS,20、50ng·ml-1)37℃孵育30min后,测定孵育后的样品与Lipid A的结合情况,以评价样品内的活性物质含量。结果在所筛选的78种中药中有12种中药的水提物与Lipid A具有较高的亲和活性,在这12种中药中有6种中药的醇提物与Lipid A也具有较高的亲和活性;通过对所筛选出的12种中药的水提物去除鞣质后与LPS的消耗实验,发现所筛选出的12种中药均含有除鞣质以外的与lipid A具有特异性结合的物质,并且12种中药中能与Lipid A发生特异性结合作用的活性物质含量存在较大的差异。结论生物传感器跟踪技术是一种快速、准确、有效的筛选平台,筛选出的12种中药均含有非鞣质类能与lipid A发生特异性结合作用的物质,为进一步对12种中药抗内毒素有效部位或单体的分离提供了重要的实验依据。

Screening of anti-endotoxin traditional Chinese medicine by target of Lipid A

AIM: To screen anti-endotoxin traditional Chinese medicine(TCM) from anti-inflammation TCM by Biosensor, and to further study anti-endotoxin effective materials of TCM. METHODS: Non-derivatised cuvette immobilised Lipid A was used to track the bonding between the Lipid A and aqueous extracts and alcoholic extracts from seventy-eight TCM, and then neutralized endotoxin in vitro by selecting hypso-bonding TCM that removed the tannins. RESULTS: Twelve aqueous extracts and six alcoholic extracts from seventy-eight TCM had hypso-bonding with Lipid A (more than 200 response unit),and twelve aqueous extracts could bind specially endotoxin after removed tannins. CONCLUSION: Track of biosensor technology is a quick, accurate and valid method in screening TCM. Aqueous extracts of twelve TCM can bind lipid A specially besides the tannins, and it is foundation to further segregate anti-endotoxin effective materials from twelve TCM in the future.