CAJ | 학술논문

目的：优化铁皮石斛多糖提取工艺,研究其对高糖诱导的静脉血管内皮细胞( HUVEC)的保护作用机制。方法在单因素实验基础上,利用响应面法优化多糖的提取工艺；体外培养HUVEC细胞,将对数生长期的细胞分为空白对照组、正常血糖组、高糖刺激组和不同铁皮石斛多糖浓度+高糖刺激组,分别作用24和48 h后,采用MTT法检测的HU-VEC凋亡的影响,用RT-PCR法检测内皮细胞Bax、Bcl-2的表达情况。结果铁皮石斛多糖提取的最优条件为：提取次数为2．8次,液(mL)料比(g)15．75∶1,提取时间2 h,在此条件下,理论计算提取率达到25．1%,实测提取率为24．9%,与模型高度拟合。与正常组相比,高糖组抑制细胞的生长,增强Bax的表达,抑制Bcl-2的表达,在高糖诱导下,铁皮石斛多糖促进细胞生长,抑制Bax的表达,增强Bcl-2的表达。结论通过响应面法优化的工艺,得率高,提取效果好。铁皮石斛多糖可通过抑制高糖诱导的血管内皮细胞Bax的表达,增强 Bcl-2的表达,从而抑制血管内皮细胞凋亡,在一定程度上可防治糖尿病血管病变。
목적：우화철피석곡다당제취공예,연구기대고당유도적정맥혈관내피세포( HUVEC)적보호작용궤제。방법재단인소실험기출상,이용향응면법우화다당적제취공예；체외배양HUVEC세포,장대수생장기적세포분위공백대조조、정상혈당조、고당자격조화불동철피석곡다당농도+고당자격조,분별작용24화48 h후,채용MTT법검측적HU-VEC조망적영향,용RT-PCR법검측내피세포Bax、Bcl-2적표체정황。결과철피석곡다당제취적최우조건위：제취차수위2．8차,액(mL)료비(g)15．75∶1,제취시간2 h,재차조건하,이론계산제취솔체도25．1%,실측제취솔위24．9%,여모형고도의합。여정상조상비,고당조억제세포적생장,증강Bax적표체,억제Bcl-2적표체,재고당유도하,철피석곡다당촉진세포생장,억제Bax적표체,증강Bcl-2적표체。결론통과향응면법우화적공예,득솔고,제취효과호。철피석곡다당가통과억제고당유도적혈관내피세포Bax적표체,증강 Bcl-2적표체,종이억제혈관내피세포조망,재일정정도상가방치당뇨병혈관병변。
Aim To optimize extraction of polysaccha-rides from Dendrobium officinale ( DOP ) and investi-gate the effects of DOP on the expression of Bax and Bcl-2 in vascular endothelial cell of human umbilical vein ( HUVEC) by the induction of high sugar. Meth-ods Response surface methodology ( RSM ) was em-ployed to optimize the conditions for extraction of DOP on the basis of single factor experiment. The HUVEC cells were divided into the blank control group, the normal blood sugar group, the high sugar stimulation group and different drug concentration group plus the high sugar stimulation group. The growth condition of cells was examined by methyl thiazolyl tetrazolium ( MTT) , and RT-PCR was used to detect the expres-sion of Bcl-2 Assaciated X protein ( Bax ) and B-cell lymphoma-2 ( Bcl-2 ) in vascular endothelial cells after 24 and 48 hours. Results The optimal extraction time was 2 h and the optimal liquid-solid ratio was 15. 75∶1 ( mL/g ) , and the optimal extraction frequency was 2. 8. Under these optimized conditions, the extraction rate of polysaccharides was 25. 1%, while the experi-mental extraction rate was 24. 9%, which was highly fit with the regression model. Compared with normal sugar group, high sugar group reduced the growth of cells and enhanced the expression of Bax, and inhibi-ted the expression of Bcl-2 . DOP promoted the growth of cells in high sugar induction, inhibited the expres-sion of Bax, and enhanced the expression of Bcl-2. Conclusions The process optimized through RSM has the extraction effect, and has certain practical signifi-cance. Through the down-regulation of the expression of Bax in vascular endothelial cells and the up-regula-tion of the expression of Bcl-2 by the induction of high sugar, DOP can restrain HUVEC, which could prevent and cure diabetic angiopathy to some extent.