CAJ | 학술논문

干细胞%干细胞与中医药%人脐血干细胞%补阳还五汤%生物碱有效部位%细胞增殖%川芎嗪%阿魏酸%静息膜电位%S期%流式细胞仪%电流钳模式榦細胞%榦細胞與中醫藥%人臍血榦細胞%補暘還五湯%生物堿有效部位%細胞增殖%川芎嗪%阿魏痠%靜息膜電位%S期%流式細胞儀%電流鉗模式
간세포%간세포여중의약%인제혈간세포%보양환오탕%생물감유효부위%세포증식%천궁진%아위산%정식막전위%S기%류식세포의%전류겸모식
stem cel s%stem cel s and traditional Chinese medicine%human umbilical cord blood stem cel s%Buyang Huanwu decoction%alkaloid%cel proliferation%tetramethylpyrazine%ferulic acid%resting membrane potential%S phase%flow cytometry%current clamp mode

背景:研究表明补阳还五汤对各种细胞均有保护作用,但机制尚不清楚,有研究推测是它的生物碱有效部位起作用.目的:观察补阳还五汤生物碱有效部位对人脐血干细胞体外增殖的影响.方法:获取临床健康产妇的新鲜脐血,制备人脐血干细胞悬液,将纯化的人脐血干细胞接种于40个培养皿中进行传代.分组:川芎嗪组给予100 g/L川芎嗪,阿魏酸组给予50 g/L阿魏酸,生物碱有效部位组(川芎嗪+阿魏酸组)给予100 g/L川芎嗪+50 g/L阿魏酸;对照组:继续用含体积分数为20%小牛血清的DMEM培养液培养.培养2,4,6,8,10 d观察人脐血干细胞增殖情况;24,72 h后流式细胞仪测定S期细胞的比例,培养10 d后电流钳模式记录细胞静息膜电位变化.结果与结论:川芎嗪组和阿魏酸组人脐血干细胞的增殖作用不明显,培养10 d时生物碱有效部位组细胞增殖情况明显高于川芎嗪组、阿魏酸组和对照组(P<0.01).生物碱有效部位组在培养24,72 h后处于S期的人脐血干细胞明显增多;川芎嗪+阿魏酸组细胞静息膜电位高于川芎嗪组、阿魏酸组和对照组(P<0.01).提示补阳还五汤生物碱有效部位能促进人脐血干细胞的体外增殖.
배경:연구표명보양환오탕대각충세포균유보호작용,단궤제상불청초,유연구추측시타적생물감유효부위기작용.목적:관찰보양환오탕생물감유효부위대인제혈간세포체외증식적영향.방법:획취림상건강산부적신선제혈,제비인제혈간세포현액,장순화적인제혈간세포접충우40개배양명중진행전대.분조:천궁진조급여100 g/L천궁진,아위산조급여50 g/L아위산,생물감유효부위조(천궁진+아위산조)급여100 g/L천궁진+50 g/L아위산;대조조:계속용함체적분수위20%소우혈청적DMEM배양액배양.배양2,4,6,8,10 d관찰인제혈간세포증식정황;24,72 h후류식세포의측정S기세포적비례,배양10 d후전류겸모식기록세포정식막전위변화.결과여결론:천궁진조화아위산조인제혈간세포적증식작용불명현,배양10 d시생물감유효부위조세포증식정황명현고우천궁진조、아위산조화대조조(P<0.01).생물감유효부위조재배양24,72 h후처우S기적인제혈간세포명현증다;천궁진+아위산조세포정식막전위고우천궁진조、아위산조화대조조(P<0.01).제시보양환오탕생물감유효부위능촉진인제혈간세포적체외증식.
BACKGROUND:Studies have shown that Buyang Huanwu decoction has protection effect on various cel s, but the mechanism is unclear. Some studies have speculated that the alkaloids of Buyang Huanwu decoction play a role. OBJECTIVE:To observe the effect of alkaloid of Buyang Huanwu decoction on the in vitro proliferation of human umbilical cord blood stem cel s. METHODS:Fresh umbilical cord blood was obtained from healthy puerperant to prepare the human umbilical cord blood stem cel s suspension, then the purified human umbilical cord blood stem cel s were seeded into 40 culture dishes for passage. The cel s were divided into four groups. The cel s in the tetramethylpyrazine group were cultured with 100 g/L tetramethylpyrazine, the cel s in the ferulic acid group were cultured with 50 g/L ferulic acid and the cel s in the alkaloids group (tetramethylpyrazine+ferulic acid group) were cultured with 100 g/L tetramethylpyrazine and 50 g/L ferulic acid;cel s in the control group were continuously cultured with Dulbecco’s modified Eagle’s medium containing fetal bovine serum with the volume fraction of 20%. The proliferation of human umbilical cord blood stem cel s were observed after cultured for 2, 4, 6, 8 and 10 days;after cultured for 24 and 72 hours, the ratio of human umbilical cord blood stem cel s in S phase was determined by flow cytometry, and after cultured for 10 days, the resting membrane potential changes were recorded with current clamp mode. RESULTS AND CONCLUSION:The proliferation of human umbilical cord blood stem cel s in the tetramethylpyrazine group and ferulic acid group was not obvious;after cultured for 10 days, the proliferation of human umbilical cord blood stem cel s in the alkaloids group was significantly higher than that in the tetramethylpyrazine group, ferulic acid group and control group (P<0.01). After cultured for 24 and 72 hours in the alkaloids group, the number of human umbilical cord blood stem cel s in S phase was increased;the resting membrane potential in the alkaloids group was higher than that in the tetramethylpyrazine group, ferulic acid group and control group (P<0.01). The results indicate that the alkaloid of Buyang Huanwu decoction can promote the proliferation of human umbilical cord blood stem cel s.