中国生化药物杂志
中國生化藥物雜誌
중국생화약물잡지
CHINESE JOURNAL OF BIOCHEMICAL PHARMACEUTICS
2014年
2期
4-6,9
,共4页
紫杉醇-壳聚糖缓释膜%胆管成纤维细胞%增殖%细胞凋亡
紫杉醇-殼聚糖緩釋膜%膽管成纖維細胞%增殖%細胞凋亡
자삼순-각취당완석막%담관성섬유세포%증식%세포조망
paclitaxel%chitosan sustain film%biliary fibroblasts cells%proliferation%apoptosis
目的：探讨紫杉醇-壳聚糖缓释膜（PTX-Chitosan Sustain film，PTX-CSF）对胆管成纤维细胞增殖和凋亡的影响。方法采用组织块法进行成纤维细胞的原代单细胞培养。将实验分为5组：未处理组，单纯PTX处理组（250 nM）和低、中、高壳聚糖PTX缓释膜处理组（100 nM、250 nM、500 nM）。使用噻唑蓝(methyl thiazolyl tetrazolium，MTT)比色法检测细胞的增殖情况，划痕实验检测紫杉醇对TGF-β1诱导细胞迁移的抑制作用，流式细胞术（flow cytometry，FCM）检测细胞的凋亡及细胞周期。结果紫杉醇-壳聚糖缓释膜能有效抑制胆管成纤维细胞的增殖，具有剂量和时间依赖性。其抑制作明显强于单纯的紫杉醇用药，且随着时间的延长，优势逐渐体现。紫杉醇-壳聚糖缓释膜能有效抑制TGF-β1对胆管成纤维细胞的促迁移作用，较单纯PTX组具有更长的时间效应。72 h时，紫杉醇-壳聚糖缓释膜组成纤维细胞的凋亡率显著高于单纯PTX组和对照组（P＜0.05），后2者之间无统计学差异。72 h时，紫杉醇-缓释膜组和单纯PTX组的G 2/M期细胞比例均较未处理组显著增加，2者相比，前者也明显高于后者，差异均具有统计学意义（P＜0.05）。结论紫杉醇-壳聚糖缓释膜具有较强的细胞毒性作用及明显的缓释效果，可以维持较长时间的有效浓度，从而较单纯紫杉醇用药更能持续地抑制胆管成纤维的增殖。
目的：探討紫杉醇-殼聚糖緩釋膜（PTX-Chitosan Sustain film，PTX-CSF）對膽管成纖維細胞增殖和凋亡的影響。方法採用組織塊法進行成纖維細胞的原代單細胞培養。將實驗分為5組：未處理組，單純PTX處理組（250 nM）和低、中、高殼聚糖PTX緩釋膜處理組（100 nM、250 nM、500 nM）。使用噻唑藍(methyl thiazolyl tetrazolium，MTT)比色法檢測細胞的增殖情況，劃痕實驗檢測紫杉醇對TGF-β1誘導細胞遷移的抑製作用，流式細胞術（flow cytometry，FCM）檢測細胞的凋亡及細胞週期。結果紫杉醇-殼聚糖緩釋膜能有效抑製膽管成纖維細胞的增殖，具有劑量和時間依賴性。其抑製作明顯彊于單純的紫杉醇用藥，且隨著時間的延長，優勢逐漸體現。紫杉醇-殼聚糖緩釋膜能有效抑製TGF-β1對膽管成纖維細胞的促遷移作用，較單純PTX組具有更長的時間效應。72 h時，紫杉醇-殼聚糖緩釋膜組成纖維細胞的凋亡率顯著高于單純PTX組和對照組（P＜0.05），後2者之間無統計學差異。72 h時，紫杉醇-緩釋膜組和單純PTX組的G 2/M期細胞比例均較未處理組顯著增加，2者相比，前者也明顯高于後者，差異均具有統計學意義（P＜0.05）。結論紫杉醇-殼聚糖緩釋膜具有較彊的細胞毒性作用及明顯的緩釋效果，可以維持較長時間的有效濃度，從而較單純紫杉醇用藥更能持續地抑製膽管成纖維的增殖。
목적：탐토자삼순-각취당완석막（PTX-Chitosan Sustain film，PTX-CSF）대담관성섬유세포증식화조망적영향。방법채용조직괴법진행성섬유세포적원대단세포배양。장실험분위5조：미처리조，단순PTX처리조（250 nM）화저、중、고각취당PTX완석막처리조（100 nM、250 nM、500 nM）。사용새서람(methyl thiazolyl tetrazolium，MTT)비색법검측세포적증식정황，화흔실험검측자삼순대TGF-β1유도세포천이적억제작용，류식세포술（flow cytometry，FCM）검측세포적조망급세포주기。결과자삼순-각취당완석막능유효억제담관성섬유세포적증식，구유제량화시간의뢰성。기억제작명현강우단순적자삼순용약，차수착시간적연장，우세축점체현。자삼순-각취당완석막능유효억제TGF-β1대담관성섬유세포적촉천이작용，교단순PTX조구유경장적시간효응。72 h시，자삼순-각취당완석막조성섬유세포적조망솔현저고우단순PTX조화대조조（P＜0.05），후2자지간무통계학차이。72 h시，자삼순-완석막조화단순PTX조적G 2/M기세포비례균교미처리조현저증가，2자상비，전자야명현고우후자，차이균구유통계학의의（P＜0.05）。결론자삼순-각취당완석막구유교강적세포독성작용급명현적완석효과，가이유지교장시간적유효농도，종이교단순자삼순용약경능지속지억제담관성섬유적증식。
Objective To explore the effect of Paclitaxel-Chitosan Sustain film on growth, apoptosis and cell cycle of biliary fibroblasts cells. Methods Human biliary fibroblasts cells were cultured and treated with PTX-CSF and naked PTX,separately, untreated cells as blank control. The experiment was divided into five groups:untreated group, simple PTX-treated group (250nM) and low, medium and high chitosan sustained-release film PTX-treated group (100 nM, 250 nM, 500 nM). The proliferations of cells were determined by MTT assay. The apoptosis and cell cycle of cells were detected by FCM. Results The proliferation of biliary fibroblasts cells was inhibited by PTX-CSF with time-dependent and dose-dependent, and the inhibiting effect was more obvious than naked PTX treatment as the time went on. Meanwhile, PTX-CSF could inhibit the magration of bile duct fibroblasts induced by TGF-β1,and had longer effect than naked PTX. After 72 h, the apoptosis rate of cells treated with PTX-CSF was significantly higher than cells treated with naked PTX or untreated cells(P<0.05), the difference between naked PTX or untreated cells was not significant. Compared with untreated cells, the proportion of G 2/M in cells treated with PTX or PTX-CSF were significantly increased, and the former was sinificantly higher than the latter(P<0.05). Conclusion Compared with naked PTX, PTX-CSF has strong cytotoxic effects and obviously sustained-release effect. The effective concentration can be maintain for a long time by PTX-CSF, and it could be as the novel drug delivery system to continuously inhibit proliferation of bile duct fibroblasts.
