中国生化药物杂志
中國生化藥物雜誌
중국생화약물잡지
CHINESE JOURNAL OF BIOCHEMICAL PHARMACEUTICS
2014年
3期
12-15
,共4页
紫杉醇%良性瘢痕纤维化%抵抗%壳寡糖缓释膜
紫杉醇%良性瘢痕纖維化%牴抗%殼寡糖緩釋膜
자삼순%량성반흔섬유화%저항%각과당완석막
paclitaxel%benign fibrosis%resistance%chitosan sustained release membrane
目的：探讨紫杉醇（paclitaxel，PTX）抗良性胆管瘢痕纤维化的最佳抵抗因素，为临床防治胆管良性瘢痕纤维化提供有效依据。方法体外培养人胆管上皮细胞（biliary epithelial cell，BEC），并将配制好的 PTX 药物浓度梯度按0.001 uM、0.005 uM、0.1 uM、0.5 uM、1 uM加入细胞培养板培养48 h，MTT法检测PTX对BEC半抑制率IC50，从而确定最佳PTX浓度；人胆管上皮细胞培养0 h，24 h，48 h，72 h，MTT 法测100 nM、250 nM、500 nM 3种不同含量的紫杉醇-壳聚糖缓释膜（PTX-Chitosan Sustained release membrane，PTX-SRM）与最佳浓度PTX对BEC的抑制率，得到最佳浓度的PTX-SRM；细胞培养48 h，72 h后采用Western Blot和Real-time PCR方法检测PTX及PTX-SRM对BECα-SMA、E-cadherin、Vimentin蛋白及基因表达变化。结果单独PTX对良性胆管瘢痕发挥抵抗作用的最佳浓度为250 nM；PTX和PTX-SRM均能有效抑制BEC的增殖、转化；中、低浓度的PTX-SRM对良性胆管瘢痕纤维化的治疗效果最佳，最佳载药量分别为100 nM、250 nM；PTX-SRM对BEC的抑制持续时间长于单独PTX（P＜0.05）。结论 PTX-SRM对BEC增殖、转化的抑制作用优于单独PTX给药，为临床对良性胆管瘢痕的预防和治疗提供了科学可行的新方法。
目的：探討紫杉醇（paclitaxel，PTX）抗良性膽管瘢痕纖維化的最佳牴抗因素，為臨床防治膽管良性瘢痕纖維化提供有效依據。方法體外培養人膽管上皮細胞（biliary epithelial cell，BEC），併將配製好的 PTX 藥物濃度梯度按0.001 uM、0.005 uM、0.1 uM、0.5 uM、1 uM加入細胞培養闆培養48 h，MTT法檢測PTX對BEC半抑製率IC50，從而確定最佳PTX濃度；人膽管上皮細胞培養0 h，24 h，48 h，72 h，MTT 法測100 nM、250 nM、500 nM 3種不同含量的紫杉醇-殼聚糖緩釋膜（PTX-Chitosan Sustained release membrane，PTX-SRM）與最佳濃度PTX對BEC的抑製率，得到最佳濃度的PTX-SRM；細胞培養48 h，72 h後採用Western Blot和Real-time PCR方法檢測PTX及PTX-SRM對BECα-SMA、E-cadherin、Vimentin蛋白及基因錶達變化。結果單獨PTX對良性膽管瘢痕髮揮牴抗作用的最佳濃度為250 nM；PTX和PTX-SRM均能有效抑製BEC的增殖、轉化；中、低濃度的PTX-SRM對良性膽管瘢痕纖維化的治療效果最佳，最佳載藥量分彆為100 nM、250 nM；PTX-SRM對BEC的抑製持續時間長于單獨PTX（P＜0.05）。結論 PTX-SRM對BEC增殖、轉化的抑製作用優于單獨PTX給藥，為臨床對良性膽管瘢痕的預防和治療提供瞭科學可行的新方法。
목적：탐토자삼순（paclitaxel，PTX）항량성담관반흔섬유화적최가저항인소，위림상방치담관량성반흔섬유화제공유효의거。방법체외배양인담관상피세포（biliary epithelial cell，BEC），병장배제호적 PTX 약물농도제도안0.001 uM、0.005 uM、0.1 uM、0.5 uM、1 uM가입세포배양판배양48 h，MTT법검측PTX대BEC반억제솔IC50，종이학정최가PTX농도；인담관상피세포배양0 h，24 h，48 h，72 h，MTT 법측100 nM、250 nM、500 nM 3충불동함량적자삼순-각취당완석막（PTX-Chitosan Sustained release membrane，PTX-SRM）여최가농도PTX대BEC적억제솔，득도최가농도적PTX-SRM；세포배양48 h，72 h후채용Western Blot화Real-time PCR방법검측PTX급PTX-SRM대BECα-SMA、E-cadherin、Vimentin단백급기인표체변화。결과단독PTX대량성담관반흔발휘저항작용적최가농도위250 nM；PTX화PTX-SRM균능유효억제BEC적증식、전화；중、저농도적PTX-SRM대량성담관반흔섬유화적치료효과최가，최가재약량분별위100 nM、250 nM；PTX-SRM대BEC적억제지속시간장우단독PTX（P＜0.05）。결론 PTX-SRM대BEC증식、전화적억제작용우우단독PTX급약，위림상대량성담관반흔적예방화치료제공료과학가행적신방법。
Objective To discuss the best resistance factors of paclitaxel(Taxinol)on benign biliary scar fibrosis,in order to provide an effective basis for clinical prevention and treatment of benign biliary scar fibrosis.Methods Human bile duct epithelial cells were cultured in vitro,the prepared PTX at 0.001 uM,0.005 uM,0.1 uM,0.5 uMand 1 uMconcentration were separately added into cells for 48 h.The half inhibitory rate of BEC (IC50) were determined by MTT and the optimal concentration were confirmed.Human bile duct epithelial cells were cultured in 0 h,24 h,48 h and 72 h,the inhibitory rate of BEC at 100 nM,250 nM,and 500 nM PTX-Chitosan Sustained release membranes and the optimal concentration of PTX were determined by MTT and the optimal concentration of PTX-SRM were obtained.Human bile duct epithelial cells were cultured for 48 h and 72 h,the mRNA and protein expression ofα-SMA,E-cadherin,Vimentin were detected by Western Blot and Real-time PCR methods.Results The optimum resistance concentration of PTX to benign biliary scar was 250 nM.PTX and PTX-SRM could effectively inhibit the proliferation and transformation of BEC,and the best effective treatments to resist benign biliary scar fibrosis were low and middle concentrations of PTX-SRM,the best drug loading were 100 nMand 250 nM.The inhibition duration of PTX-SRMon BEC was longer than PTX alone(P<0.05).Conclusion The inhibition of PTX-SRMon BEC proliferation and transformation is better than the single drug of PTX,which provides a new scientific and feasible method for clinical prevention and treatment of benign biliary scar.
