中国现代医学杂志
中國現代醫學雜誌
중국현대의학잡지
CHINA JOURNAL OF MODERN MEDICINE
2008年
5期
513-517
,共5页
热休克蛋白%NK细胞%小鼠%脾脏%单个核细胞
熱休剋蛋白%NK細胞%小鼠%脾髒%單箇覈細胞
열휴극단백%NK세포%소서%비장%단개핵세포
HSP%NK cell%mouse%spleen%MNBCCLC nulnber:R-332 Document code:A
目的 观察皮下注射重组可溶性融合小鼠热休克蛋白70(rHSP70)后对小鼠脾脏中NK细胞在单核细胞中所占比例的影响.方法 取C57/BL6小鼠18只,随机等分3组.第1组每只在颈部皮下注射20ìgrHSP70(高剂量组),第2组每只注射10ìg(低剂量组),第3组注射磷酸钠缓冲液(对照组),注射体积均为每只200ìL.1周1次,共2次.第3周杀鼠,从脾分离其单个核细胞(MNBC),以藻红蛋白NK1.1单抗荧光标记后,在流式细胞仪上测定NK细胞及单个核细胞总数,计算NK细胞的比例.结果 NK细胞数在高剂量组、低剂量组和对照组中的比例分别占(12.7±1.2)%、(34.3±11.3)%和(10.0±0.8)%,经方差分析,高剂量组与低剂量组间,以及低剂量组与对照组组间在统计学上均差异非常显著(p<0.01),其小鼠脾脏中NK细胞HUANG Wei-da:Author to whom conespondence should be addressed.Telephone and Fax:+86-021-55522773.E-mail:whuang@fudan.edu.cn占MNBC之比例增加了2倍多.而3组的脾脏重量分别为(0.086±0.004)、(0.090±0.009)和(0.087±0.006)g,各组问差异均无统计学意义(P>0.05).结论 低剂量小鼠rHSP70可大幅提高小鼠脾脏NK细胞在MNBC中的比例,它有可能成为一种免疫治疗的潜在药物.每只20ìg的高剂量反而不如10ìg低剂量的促进效果明显.提示rHSP70对小鼠牌脏NK细胞的调剂存在双向模式.
目的 觀察皮下註射重組可溶性融閤小鼠熱休剋蛋白70(rHSP70)後對小鼠脾髒中NK細胞在單覈細胞中所佔比例的影響.方法 取C57/BL6小鼠18隻,隨機等分3組.第1組每隻在頸部皮下註射20ìgrHSP70(高劑量組),第2組每隻註射10ìg(低劑量組),第3組註射燐痠鈉緩遲液(對照組),註射體積均為每隻200ìL.1週1次,共2次.第3週殺鼠,從脾分離其單箇覈細胞(MNBC),以藻紅蛋白NK1.1單抗熒光標記後,在流式細胞儀上測定NK細胞及單箇覈細胞總數,計算NK細胞的比例.結果 NK細胞數在高劑量組、低劑量組和對照組中的比例分彆佔(12.7±1.2)%、(34.3±11.3)%和(10.0±0.8)%,經方差分析,高劑量組與低劑量組間,以及低劑量組與對照組組間在統計學上均差異非常顯著(p<0.01),其小鼠脾髒中NK細胞HUANG Wei-da:Author to whom conespondence should be addressed.Telephone and Fax:+86-021-55522773.E-mail:whuang@fudan.edu.cn佔MNBC之比例增加瞭2倍多.而3組的脾髒重量分彆為(0.086±0.004)、(0.090±0.009)和(0.087±0.006)g,各組問差異均無統計學意義(P>0.05).結論 低劑量小鼠rHSP70可大幅提高小鼠脾髒NK細胞在MNBC中的比例,它有可能成為一種免疫治療的潛在藥物.每隻20ìg的高劑量反而不如10ìg低劑量的促進效果明顯.提示rHSP70對小鼠牌髒NK細胞的調劑存在雙嚮模式.
목적 관찰피하주사중조가용성융합소서열휴극단백70(rHSP70)후대소서비장중NK세포재단핵세포중소점비례적영향.방법 취C57/BL6소서18지,수궤등분3조.제1조매지재경부피하주사20ìgrHSP70(고제량조),제2조매지주사10ìg(저제량조),제3조주사린산납완충액(대조조),주사체적균위매지200ìL.1주1차,공2차.제3주살서,종비분리기단개핵세포(MNBC),이조홍단백NK1.1단항형광표기후,재류식세포의상측정NK세포급단개핵세포총수,계산NK세포적비례.결과 NK세포수재고제량조、저제량조화대조조중적비례분별점(12.7±1.2)%、(34.3±11.3)%화(10.0±0.8)%,경방차분석,고제량조여저제량조간,이급저제량조여대조조조간재통계학상균차이비상현저(p<0.01),기소서비장중NK세포HUANG Wei-da:Author to whom conespondence should be addressed.Telephone and Fax:+86-021-55522773.E-mail:whuang@fudan.edu.cn점MNBC지비례증가료2배다.이3조적비장중량분별위(0.086±0.004)、(0.090±0.009)화(0.087±0.006)g,각조문차이균무통계학의의(P>0.05).결론 저제량소서rHSP70가대폭제고소서비장NK세포재MNBC중적비례,타유가능성위일충면역치료적잠재약물.매지20ìg적고제량반이불여10ìg저제량적촉진효과명현.제시rHSP70대소서패장NK세포적조제존재쌍향모식.
[Objective]To construct the recombinant heat shock protein 70(rHSP70)expression plasmid(pET-28b-HSP70),express and purify the rHSP70,and finally investigate the effect of soluble routine rHSP70 on splenic natural killer(NK)cells in mice.[Methods]Eighteen mice(C57BL/6)were randomly divided into three groups with six mice in each:High dose group(HDG),low dose group(LDC)and control group(CG).The mice in HDO,LDG spleens were all weighed,mononuclear blood cells(MNBCs)were also isolated from spleen and directly stained with phycoerythrin(PE)-conjugated monoclonal anti-NK1.1 antibody.The NK cells and MNBCs were both assayed with flow cytometry.[Results]The average proportions of splenic NK cells of HDG,LDG and CG were(12.7+1.2)%,(34.3+11.3)% and(10.0+0.8)%,respectively.There were significant differences between the low dose group LDG and each ofthe remaining two groups(both P＜0.01).The weights of spleens of three groups were(0.086+0.004),(0.090+0.009)and(0.087+0.006)g,respectively,There were no differences between them(P＞0.05).[Conclusion]The results suggest that low dose of murine rHSP70 may increase the subpopalation of NK cells in murine spleen,which might be a potential biological regulation drug.The observation that the high dose of rHSPT0 does not have so remarkable promoting effect on proportions of splenic NK cells in mice as the low dose suggests that there are bidirectional regulation effects of rHSP70 on splenic NK cells.
