中国医药导报
中國醫藥導報
중국의약도보
CHINA MEDICAL HERALD
2014年
18期
17-21,26
,共6页
崔雅璠%丁樱%杨晓青%段凤阳%翟宗岗%刘丽雅
崔雅璠%丁櫻%楊曉青%段鳳暘%翟宗崗%劉麗雅
최아번%정앵%양효청%단봉양%적종강%류려아
紫癜性肾炎%IgA1%系膜细胞%肿瘤坏死因子-α
紫癜性腎炎%IgA1%繫膜細胞%腫瘤壞死因子-α
자전성신염%IgA1%계막세포%종류배사인자-α
Henoch-schonlein purpura nephritis (HSPN)%IgA1%Mesangial cell%Tumor necrosis factor-α
目的：观察过敏性紫癜性肾炎(HSPN)患儿及健康儿童血清IgA1对小鼠肾小球系膜细胞(MES)增殖及TNF-α分泌的影响.方法采集2013年5~12月于河南中医学院第一附属医院儿科住院的40例HSPN患儿及20名健康儿童血清；通过Jacalin亲和层析联合Superdex-200分子筛层析的方法将血清中单聚体IgA1(mIgA1)纯化、分离,将mIgA1热聚合为聚合IgA1(aIgA1)；采用不同浓度的患儿及健康儿童的aIgA1分别刺激MES细胞株,用MTT法检测24 h及48 h的细胞增殖情况；收集培养48 h的细胞培养基上清,用ELISA法测定TNF-α的水平.结果在HSPN患儿aIgA1刺激结果中,不同浓度aIgA1刺激组间MTT实验吸光度的比较,差异有统计学意义(P<0.05),24 h MTT实验结果显示院与阴性对照比较,aIgA1刺激浓度在250、1000μg/mL时,细胞增殖差异均有统计学意义(P<0.05),在aIgA1浓度为500和750μg/mL时,细胞增殖差异有高度统计学意义(P<0.01)；48 h MTT实验结果显示院与阴性对照比较,在aIgA1刺激浓度50μg/mL时,细胞增殖差异有统计学意义(P<0.05),在100~1000μg/mL时,细胞增殖差异均有高度统计学意义(P<0.01).健康儿童在aIgA1刺激浓度在750和1000μg/mL可刺激MES增殖,与阴性对照比较,差异有统计学意义(P<0.05). HSPN患儿aIgA1可刺激MES分泌TNF-α水平升高,与阴性对照比较,差异有高度统计学意义(P<0.01),且随aIgA1浓度增加呈上升趋势.结论 HSPN患儿aIgA1可刺激MES增殖及分泌TNF-α水平增加,且这一结果具有浓度±赖性及饱和性。
目的：觀察過敏性紫癜性腎炎(HSPN)患兒及健康兒童血清IgA1對小鼠腎小毬繫膜細胞(MES)增殖及TNF-α分泌的影響.方法採集2013年5~12月于河南中醫學院第一附屬醫院兒科住院的40例HSPN患兒及20名健康兒童血清；通過Jacalin親和層析聯閤Superdex-200分子篩層析的方法將血清中單聚體IgA1(mIgA1)純化、分離,將mIgA1熱聚閤為聚閤IgA1(aIgA1)；採用不同濃度的患兒及健康兒童的aIgA1分彆刺激MES細胞株,用MTT法檢測24 h及48 h的細胞增殖情況；收集培養48 h的細胞培養基上清,用ELISA法測定TNF-α的水平.結果在HSPN患兒aIgA1刺激結果中,不同濃度aIgA1刺激組間MTT實驗吸光度的比較,差異有統計學意義(P<0.05),24 h MTT實驗結果顯示院與陰性對照比較,aIgA1刺激濃度在250、1000μg/mL時,細胞增殖差異均有統計學意義(P<0.05),在aIgA1濃度為500和750μg/mL時,細胞增殖差異有高度統計學意義(P<0.01)；48 h MTT實驗結果顯示院與陰性對照比較,在aIgA1刺激濃度50μg/mL時,細胞增殖差異有統計學意義(P<0.05),在100~1000μg/mL時,細胞增殖差異均有高度統計學意義(P<0.01).健康兒童在aIgA1刺激濃度在750和1000μg/mL可刺激MES增殖,與陰性對照比較,差異有統計學意義(P<0.05). HSPN患兒aIgA1可刺激MES分泌TNF-α水平升高,與陰性對照比較,差異有高度統計學意義(P<0.01),且隨aIgA1濃度增加呈上升趨勢.結論 HSPN患兒aIgA1可刺激MES增殖及分泌TNF-α水平增加,且這一結果具有濃度±賴性及飽和性。
목적：관찰과민성자전성신염(HSPN)환인급건강인동혈청IgA1대소서신소구계막세포(MES)증식급TNF-α분비적영향.방법채집2013년5~12월우하남중의학원제일부속의원인과주원적40례HSPN환인급20명건강인동혈청；통과Jacalin친화층석연합Superdex-200분자사층석적방법장혈청중단취체IgA1(mIgA1)순화、분리,장mIgA1열취합위취합IgA1(aIgA1)；채용불동농도적환인급건강인동적aIgA1분별자격MES세포주,용MTT법검측24 h급48 h적세포증식정황；수집배양48 h적세포배양기상청,용ELISA법측정TNF-α적수평.결과재HSPN환인aIgA1자격결과중,불동농도aIgA1자격조간MTT실험흡광도적비교,차이유통계학의의(P<0.05),24 h MTT실험결과현시원여음성대조비교,aIgA1자격농도재250、1000μg/mL시,세포증식차이균유통계학의의(P<0.05),재aIgA1농도위500화750μg/mL시,세포증식차이유고도통계학의의(P<0.01)；48 h MTT실험결과현시원여음성대조비교,재aIgA1자격농도50μg/mL시,세포증식차이유통계학의의(P<0.05),재100~1000μg/mL시,세포증식차이균유고도통계학의의(P<0.01).건강인동재aIgA1자격농도재750화1000μg/mL가자격MES증식,여음성대조비교,차이유통계학의의(P<0.05). HSPN환인aIgA1가자격MES분비TNF-α수평승고,여음성대조비교,차이유고도통계학의의(P<0.01),차수aIgA1농도증가정상승추세.결론 HSPN환인aIgA1가자격MES증식급분비TNF-α수평증가,차저일결과구유농도±뢰성급포화성。
Objective To explore the effects of IgA1 isolated from both children patients with HSPN and healthy con-trols on proliferation and the level of TNF-α of MES. Methods Serum of 40 children with HSPN and 20 healthy chil-dren in the First Affiliated Hospital of He'nan University of TCM were collected from May to December 2013. IgA1 was purified from by Jacalin affinity chromatography and molecular sieve chromatography, then aggregated into aIgA1. MES was stimulated by different concentrations of aIgA1 with both children patients of HSPN and healthy controls, and the effects of different concentration aIgA1 on cells proliferation and the changes of secretion level for TNF-αwere ob-served and contrasted. Results By the IgA1 stimulating of children patients with HSPN, there were statistical differ-ences among the MTT absorbency of mesangial cell in different concentration stimulation of aIgA1 (P< 0.05). Com-pared with negative control, the absorbency had statistically significant differences at the concentration of aIgA1 from 250 μg/mL to 1000 μg/mL of HSPN childern (P< 0.05), and the absorbency had high statistical significances at the concentration of aIgA1 from 500 μg/mL to 750μg/mL of HSPN children at 24 h (P< 0.01). Compared with nega-tive control, the absorbency had statistically significant difference at the concentration of aIgA1 in 50 μg/mL of HSPN children (P<0.05), and the absorbency had high statistical significances at the concentration of aIgA1 from 100 to 1000μg/mL of HSPN children at 48 h (P<0.01). Compared with negative control, the absorbency had sta-tistically significant differences at the concentration of aIgA1 from 750μg/mL to 1000μg/mL of healthy children (P<0.05). Compared with negative control, aIgA1 of children patients with HSPN could stimulate the secretion of TNF-α, and the level of TNF-α of HSPN chiuldren had high statistical significances (P < 0.01). Conclusion The aIgA1 of children patients with HSPN can stimulate the cells proliferation and increase the secretion of TNF-α, and this result has the concentration-dependent and saturation.
