中华内分泌外科杂志
中華內分泌外科雜誌
중화내분비외과잡지
CHINESE JOURNAL OF ENDOCRINE SURGERY
2014年
6期
447-451
,共5页
刘柳%张雪%张文涛%宁淑华
劉柳%張雪%張文濤%寧淑華
류류%장설%장문도%저숙화
胚胎成纤维细胞%糖尿病溃疡%皮肤组织工程%胶原代谢
胚胎成纖維細胞%糖尿病潰瘍%皮膚組織工程%膠原代謝
배태성섬유세포%당뇨병궤양%피부조직공정%효원대사
Embryonic fibroblasts%Diabetic ulcer%Tissue engineered skin%Collagen metabolism
目的：观察人胚胎成纤维细胞（ FBs）对糖尿病足FBs生物学特性的影响，探讨其促进糖尿病溃疡愈合的机制。方法分离培养糖尿病创面FBs10例，分别与孕中期人胚胎皮肤FBs 3例（实验组）、正常人皮肤FBs 5例（对照组1）于transwell小室培养体系中间接共培养，并设立空白对照（对照组2）。绘制共培养体系中糖尿病足FBs生长曲线；观察共培养7 d后糖尿病足FBs形态；共培养至第4、7 d后，将各组糖尿病足FBs分离培养2 d，测量和对比上清液中羟脯氨酸和TGF-β1含量；检测共培养第3、5、7 d，β-半乳糖苷酶（ SA-β-gal ）染色阳性细胞百分率。胚胎FBs制作三维培养应用于8例糖尿病足溃疡，比较治疗前后创面愈合时间，做自身前后对照研究。结果实验组糖尿病足FBs比2个对照组增殖活跃，形态更接近于正常。经胚胎FBs处理4、7 d，糖尿病足FBs培养上清液中羟脯氨酸、TGF-β1明显增高，相对2个对照组差异均有统计学意义（羟脯氨酸4、7 d：与对照组1相比P＝0.023、P＝0.007；与对照组2相比P＝0.007、P＝0.003；TGF-β14、7 d：与对照组1相比P＝0.000、P＝0.000；与对照组2相比P＝0.000、P＝0.000）。实验组糖尿病足FBs的SA-β-gal染色阳性率在共培养第3、5及7 d无明显变化，2个对照组则明显增高。8例临床糖尿病足溃疡应用胚胎FBs三维培养治疗后创面均愈合。结论胚胎FBs能显著促进糖尿病足FBs增殖，延缓老化表型出现，同时上调羟脯氨酸、TGF-β1的分泌，可能与促进创面愈合机制有关。
目的：觀察人胚胎成纖維細胞（ FBs）對糖尿病足FBs生物學特性的影響，探討其促進糖尿病潰瘍愈閤的機製。方法分離培養糖尿病創麵FBs10例，分彆與孕中期人胚胎皮膚FBs 3例（實驗組）、正常人皮膚FBs 5例（對照組1）于transwell小室培養體繫中間接共培養，併設立空白對照（對照組2）。繪製共培養體繫中糖尿病足FBs生長麯線；觀察共培養7 d後糖尿病足FBs形態；共培養至第4、7 d後，將各組糖尿病足FBs分離培養2 d，測量和對比上清液中羥脯氨痠和TGF-β1含量；檢測共培養第3、5、7 d，β-半乳糖苷酶（ SA-β-gal ）染色暘性細胞百分率。胚胎FBs製作三維培養應用于8例糖尿病足潰瘍，比較治療前後創麵愈閤時間，做自身前後對照研究。結果實驗組糖尿病足FBs比2箇對照組增殖活躍，形態更接近于正常。經胚胎FBs處理4、7 d，糖尿病足FBs培養上清液中羥脯氨痠、TGF-β1明顯增高，相對2箇對照組差異均有統計學意義（羥脯氨痠4、7 d：與對照組1相比P＝0.023、P＝0.007；與對照組2相比P＝0.007、P＝0.003；TGF-β14、7 d：與對照組1相比P＝0.000、P＝0.000；與對照組2相比P＝0.000、P＝0.000）。實驗組糖尿病足FBs的SA-β-gal染色暘性率在共培養第3、5及7 d無明顯變化，2箇對照組則明顯增高。8例臨床糖尿病足潰瘍應用胚胎FBs三維培養治療後創麵均愈閤。結論胚胎FBs能顯著促進糖尿病足FBs增殖，延緩老化錶型齣現，同時上調羥脯氨痠、TGF-β1的分泌，可能與促進創麵愈閤機製有關。
목적：관찰인배태성섬유세포（ FBs）대당뇨병족FBs생물학특성적영향，탐토기촉진당뇨병궤양유합적궤제。방법분리배양당뇨병창면FBs10례，분별여잉중기인배태피부FBs 3례（실험조）、정상인피부FBs 5례（대조조1）우transwell소실배양체계중간접공배양，병설립공백대조（대조조2）。회제공배양체계중당뇨병족FBs생장곡선；관찰공배양7 d후당뇨병족FBs형태；공배양지제4、7 d후，장각조당뇨병족FBs분리배양2 d，측량화대비상청액중간포안산화TGF-β1함량；검측공배양제3、5、7 d，β-반유당감매（ SA-β-gal ）염색양성세포백분솔。배태FBs제작삼유배양응용우8례당뇨병족궤양，비교치료전후창면유합시간，주자신전후대조연구。결과실험조당뇨병족FBs비2개대조조증식활약，형태경접근우정상。경배태FBs처리4、7 d，당뇨병족FBs배양상청액중간포안산、TGF-β1명현증고，상대2개대조조차이균유통계학의의（간포안산4、7 d：여대조조1상비P＝0.023、P＝0.007；여대조조2상비P＝0.007、P＝0.003；TGF-β14、7 d：여대조조1상비P＝0.000、P＝0.000；여대조조2상비P＝0.000、P＝0.000）。실험조당뇨병족FBs적SA-β-gal염색양성솔재공배양제3、5급7 d무명현변화，2개대조조칙명현증고。8례림상당뇨병족궤양응용배태FBs삼유배양치료후창면균유합。결론배태FBs능현저촉진당뇨병족FBs증식，연완노화표형출현，동시상조간포안산、TGF-β1적분비，가능여촉진창면유합궤제유관。
Objective To observe the effects of human embryonic fibroblasts ( FBs ) on the biological characteristics of diabetic FBs in vitro , and to explore the possible therapeutic mechanism on diabetic ulcers . Methods Diabetic FBs were isolated and cultured with embryonic FBs (experimental group), adult FBs(control group 1)and alone(control group 2)in transwell chambers.Diabetic FBs growth curves in the coculture systems were described.The cell shape was observed by microscopy with HE staining .On the 4th and 7th day of cocul-ture, diabetic FBs were isolated to measure the expression levels of hydroxyproline and transforming growth factor -beta 1(TGF-β1)in the supernatant of monoculture .The cell senescense was displayed by β-galactosidase histo-chemical staining on the 3th, 5th and 7th day of coculture.Eight patients with diabetic foot ulcers who were can-didates for embryonic FBs constructs after failed traditional treatment were included in the study .The pre and post self-control study was conducted by observing the wound healing process before and after embryonic FBs treat -ment.Results Diabetic FBs in the experimental coculture system displayed more typical pattern and faster grow -ing rate.The level of hydroxyproline and TGF-β1 in the supernatant of diabetic FBs monoculture was significantly higher in the experimental group than in the control group after treated 4 or 7 days ( hydroxyproline:4 d, 7 d :compared with control group 1:P=0.023,P=0.007;compared with control group 2: P=0.007, P=0.003;TGF-β1 4 d, 7 d:compared with control group 1:P=0.000, P=0.000;compared with control group 2: P=0.000, P=0.000).The expression of SA-β-gal tended to be more apparent in control group 1 and 2 than in the experimental group on the 3rd,5th,and 7th day.All of the 8 cases with diabetic ulcers were cured by using em-bryonic FBs constructs .Conclusion Embryonic FBs can promote the proliferation of diabetic FBs , correct cell senescence and increase the production of hydroxyproline and TGF-β1,which maybe enhance the process of dia-betic wound healing .
