中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2013年
28期
2225-2229
,共5页
童瑾%周向东%尤列·皮尔曼%维克多·科罗索夫
童瑾%週嚮東%尤列·皮爾曼%維剋多·科囉索伕
동근%주향동%우렬·피이만%유극다·과라색부
甘草酸%醛还原酶%白细胞介素13%黏蛋白类
甘草痠%醛還原酶%白細胞介素13%黏蛋白類
감초산%철환원매%백세포개소13%점단백류
Glycyrrhizic acid%Aldehyde reductase%Interleukin-13%Mucins
目的 探讨甘草酸对白细胞介素13(IL-13)诱导的气道黏液高分泌的作用.方法 SD大鼠50只用随机数字表法随机分为5组:对照组、IL-13组和不同浓度(25、50、75 mg/kg)甘草酸组.通过黏液组化染色,采用形态定量法计算气道上皮组织染色阳性部分表达强度积分；体外培养人支气管上皮细胞株HBE-16细胞,分为6组:阴性对照组(生理盐水)、IL-13对照组(10 μg/L的IL-13+生理盐水)和不同浓度甘草酸干预组(10 μg/L的IL-13分别加25、50、75 μmol/L甘草酸)、阳性对照组(10 μg/L的IL-13 +25 μmol/L唑泊司他),通过反转录-PCR、Western印迹、荧光法和活性氧细胞渗透性指示剂(CM-H2DCFDA)探针的荧光强度分别检测各组HBE-16细胞黏蛋白(MUC) 5AC mRNA、MUC5AC蛋白表达及细胞内的醛糖还原酶(AR)活性和活性氧(ROS)相对表达水平.结果 体内实验:各组气道上皮组织染色阳性细胞表达强度积分分别为0.12±0.03、0.87±0.13、0.56±0.08、0.46±0.06、0.35±0.04,不同浓度甘草酸组明显低于IL-13组(均P＜O.05),并有剂量依赖性；IL-13组明显强于对照组(P＜0.05).体外实验:各组HBE16细胞的AR活性及48 h时的ROS相对表达水平分别为0.156±0.021、0.692±0.039、0.436±0.019、0.323±0.042、0.290±0.027和5.127±0.033、24.257±3.263、11.966±0.283、8.892±0.521、6.426±0.173,IL-13对照组明显高于阴性对照组(P＜0.05),不同浓度甘草酸干预组明显低于IL-13对照组(均P＜0.05)；各组HBE16细胞MUC5AC mRNA和MUC5AC蛋白表达分别为0.82±0.05、3.22±0.12、2.57±0.34、2.09±0.54、1.58±0.22和0.18±0.04、0.65±0.15、0.48±0.11、0.33±0.19、0.26±0.06,IL-13对照组明显高于阴性对照组(P＜0.05),不同浓度甘草酸干预组明显低于IL-13对照组(均P＜0.05).结论 甘草酸可抑制IL-13诱导的MUC5AC mRNA和蛋白质的表达,抑制气道黏液高分泌.
目的 探討甘草痠對白細胞介素13(IL-13)誘導的氣道黏液高分泌的作用.方法 SD大鼠50隻用隨機數字錶法隨機分為5組:對照組、IL-13組和不同濃度(25、50、75 mg/kg)甘草痠組.通過黏液組化染色,採用形態定量法計算氣道上皮組織染色暘性部分錶達彊度積分；體外培養人支氣管上皮細胞株HBE-16細胞,分為6組:陰性對照組(生理鹽水)、IL-13對照組(10 μg/L的IL-13+生理鹽水)和不同濃度甘草痠榦預組(10 μg/L的IL-13分彆加25、50、75 μmol/L甘草痠)、暘性對照組(10 μg/L的IL-13 +25 μmol/L唑泊司他),通過反轉錄-PCR、Western印跡、熒光法和活性氧細胞滲透性指示劑(CM-H2DCFDA)探針的熒光彊度分彆檢測各組HBE-16細胞黏蛋白(MUC) 5AC mRNA、MUC5AC蛋白錶達及細胞內的醛糖還原酶(AR)活性和活性氧(ROS)相對錶達水平.結果 體內實驗:各組氣道上皮組織染色暘性細胞錶達彊度積分分彆為0.12±0.03、0.87±0.13、0.56±0.08、0.46±0.06、0.35±0.04,不同濃度甘草痠組明顯低于IL-13組(均P＜O.05),併有劑量依賴性；IL-13組明顯彊于對照組(P＜0.05).體外實驗:各組HBE16細胞的AR活性及48 h時的ROS相對錶達水平分彆為0.156±0.021、0.692±0.039、0.436±0.019、0.323±0.042、0.290±0.027和5.127±0.033、24.257±3.263、11.966±0.283、8.892±0.521、6.426±0.173,IL-13對照組明顯高于陰性對照組(P＜0.05),不同濃度甘草痠榦預組明顯低于IL-13對照組(均P＜0.05)；各組HBE16細胞MUC5AC mRNA和MUC5AC蛋白錶達分彆為0.82±0.05、3.22±0.12、2.57±0.34、2.09±0.54、1.58±0.22和0.18±0.04、0.65±0.15、0.48±0.11、0.33±0.19、0.26±0.06,IL-13對照組明顯高于陰性對照組(P＜0.05),不同濃度甘草痠榦預組明顯低于IL-13對照組(均P＜0.05).結論 甘草痠可抑製IL-13誘導的MUC5AC mRNA和蛋白質的錶達,抑製氣道黏液高分泌.
목적 탐토감초산대백세포개소13(IL-13)유도적기도점액고분비적작용.방법 SD대서50지용수궤수자표법수궤분위5조:대조조、IL-13조화불동농도(25、50、75 mg/kg)감초산조.통과점액조화염색,채용형태정량법계산기도상피조직염색양성부분표체강도적분；체외배양인지기관상피세포주HBE-16세포,분위6조:음성대조조(생리염수)、IL-13대조조(10 μg/L적IL-13+생리염수)화불동농도감초산간예조(10 μg/L적IL-13분별가25、50、75 μmol/L감초산)、양성대조조(10 μg/L적IL-13 +25 μmol/L서박사타),통과반전록-PCR、Western인적、형광법화활성양세포삼투성지시제(CM-H2DCFDA)탐침적형광강도분별검측각조HBE-16세포점단백(MUC) 5AC mRNA、MUC5AC단백표체급세포내적철당환원매(AR)활성화활성양(ROS)상대표체수평.결과 체내실험:각조기도상피조직염색양성세포표체강도적분분별위0.12±0.03、0.87±0.13、0.56±0.08、0.46±0.06、0.35±0.04,불동농도감초산조명현저우IL-13조(균P＜O.05),병유제량의뢰성；IL-13조명현강우대조조(P＜0.05).체외실험:각조HBE16세포적AR활성급48 h시적ROS상대표체수평분별위0.156±0.021、0.692±0.039、0.436±0.019、0.323±0.042、0.290±0.027화5.127±0.033、24.257±3.263、11.966±0.283、8.892±0.521、6.426±0.173,IL-13대조조명현고우음성대조조(P＜0.05),불동농도감초산간예조명현저우IL-13대조조(균P＜0.05)；각조HBE16세포MUC5AC mRNA화MUC5AC단백표체분별위0.82±0.05、3.22±0.12、2.57±0.34、2.09±0.54、1.58±0.22화0.18±0.04、0.65±0.15、0.48±0.11、0.33±0.19、0.26±0.06,IL-13대조조명현고우음성대조조(P＜0.05),불동농도감초산간예조명현저우IL-13대조조(균P＜0.05).결론 감초산가억제IL-13유도적MUC5AC mRNA화단백질적표체,억제기도점액고분비.
Objective To explore the effects of glycyrrhizin on airway mucus hypersecretion induced by interleukin-13 (IL-13) in rats.Methods A total of 50 SD rats were divided randomly into 5 groups with a random digit table:control group,IL-13 group,and different dosage (25,50,75 mg/kg) glycyrrhizin groups.The integral of expression intensity in positive cells of airway epithelium under mucus histochemical stain was calculated with modality-quantitative method.HBE-16 cells were divided into 6 groups:negative control (physiological saline),IL-13 control (10 μg/L IL-13 + physiological saline),different concentration glycyrrhizin interference (10 μg/L IL-13 + 25,50 and 75 μmoL/L glycyrrhizin,respectively) and positive control (10 μg/L IL-13 + 25 μmol/L zopolrestat).The expression of mucin (MUC) 5AC mRNA,MUC5AC protein,aldose reductase (AR) activity and reactive oxygen species (ROS) content were detected by reverse transcription-polymerase chain reaction (RT-PCR),Western blot,fluorometric method and fluorescence intensity with General Oxidative Stress Indicator (CM-H2DFDA) catheter respectively.Results In vivo,the integral of expression intensity in positive stain cells of airway epithelium were 0.12 ± 0.03,0.87 ± 0.13,0.56 ± 0.08,0.46 ±0.06 and 0.35 ±0.04 respectively while the integral of different dosage glycyrrhizin groups was significantly lower than that of IL-13 group (all P ＜O.05) with dose depentency and the IL-13 group was stronger than control group (P ＜0.05).In vitro,the index of AR activity and ROS at 48 h of HBE16 cells in every group were 0.156 ±0.021,0.692 ±0.039,0.436±0.019,0.323 ±0.042 and 0.290 ±0.027; 5.127 ±0.033,24.257 ±3.263,11.966 ±0.283,8.892 ±0.521 and 6.426 ±0.173 respectively.The indices of IL-13 control group were higher than those of negative control group (P ＜ 0.05) while those of different concentration glycyrrhizin interference groups were lower than those of IL-13 control group (all P ＜0.05).The expressions of MUCSAC mRNA and protein of HBE16 cells in every group were 0.82 ±0.05,3.22 ±0.12,2.57 ±0.34,2.09 ±0.54 and 1.58 ±0.22;0.18 ±0.04,0.65 ±0.15,0.48 ±0.11,0.33 ±0.19 and 0.26 ±0.06 respectively.The indices of IL-13 control group were higher than those of negative control group (P ＜ 0.05) and those of different concentration glycyrrhizin interference groups were lower than those of IL-13 control group (P ＜ 0.05).Conclusion Glycyrrhizin may inhibit the expression of MUC5AC mRNA and MUC5AC protein induced by IL-13 and control the hypersecretion of airway mucus.
