CAJ | 학술논문

目的：我们在研究杞金方药毒理学及安全性的基础上，观察以杞金方药（由枸杞子、鸡内金、焦山楂、木瓜等药物组成）对地鼠肾细胞生长及早期铅干预对新生儿神经行为的影响，研究其对细胞生长及早期铅干预对新生儿神经行为的发育。方法将生长状态良好的地鼠肾细胞用Versence-胰酶混合液消化后制成4×105浓度的细胞悬液接种24孔细胞培养板，每孔1mL，37℃二氧化碳培养。称取杞金方药颗粒剂10g放入100mL配液瓶中，加入50mL细胞培养液，121℃高压20min，4℃冰箱放置24h。不断搅拌，并使未溶解部分沉淀。同时将苯酚按同样方法处理做为毒性对照组。长成单层的地鼠肾细胞抽出生长液，将制备好的浸提液用细胞培养液1∶2~1∶32稀释后加入各孔，每稀释度接种2孔，同时设细胞对照和毒性对照。次日开始观察细胞生长状态。连续观察5d。受试产妇40例，受试组每d1剂，煎汤分3~5次服，10d为1疗程。该方药是由鸡内金10~15份、构杞子10~15份、焦山楂5~10份、木瓜5~10份组成。将乳汁铅水平≥0.24μmol/L和＜0.24μmol/L设为高铅组和低铅组。受试前后两组间乳汁铅水平进行比较，在3个月龄时对研究对象用蓓蕾（Bayley）婴儿发育量表进行发育评价3个月±1周婴儿精神发育指数（MDI）和心理运动发育指数（PDI）。石墨炉原子吸收光谱法测定初乳铅含量。结果杞金方药制备的颗粒按不同稀释倍数接种于生长良好的地鼠肾细胞观察其对细胞生长的影响，经36℃连续观察5d未见细胞固缩、肿胀、脱落等细胞毒性反应，证实杞金方药不会引起对细胞的毒性反应。受试组和对照组服药前后进行乳铅水平比较，受试组乳铅水平明显低于对照组，两者比较差异有显著性。40例婴儿3个月龄时MDI、PDI与杞金方药合剂干预的相关回归分析结果表明，两发育指标和杞金方药合剂早期铅干预间存在有统计学意义的负相关。结论供试液按不同稀释倍数接种于生长良好的单层地鼠肾细胞，观察其对细胞生长的影响，单层地鼠肾细胞生长良好，与对照组比较细胞生长无差异，对地鼠肾细胞生长无不良影响。用该方药合剂早期干预初乳铅水平，对新生儿神经发育有一定影响。目的：我們在研究杞金方藥毒理學及安全性的基礎上，觀察以杞金方藥（由枸杞子、鷄內金、焦山楂、木瓜等藥物組成）對地鼠腎細胞生長及早期鉛榦預對新生兒神經行為的影響，研究其對細胞生長及早期鉛榦預對新生兒神經行為的髮育。方法將生長狀態良好的地鼠腎細胞用Versence-胰酶混閤液消化後製成4×105濃度的細胞懸液接種24孔細胞培養闆，每孔1mL，37℃二氧化碳培養。稱取杞金方藥顆粒劑10g放入100mL配液瓶中，加入50mL細胞培養液，121℃高壓20min，4℃冰箱放置24h。不斷攪拌，併使未溶解部分沉澱。同時將苯酚按同樣方法處理做為毒性對照組。長成單層的地鼠腎細胞抽齣生長液，將製備好的浸提液用細胞培養液1∶2~1∶32稀釋後加入各孔，每稀釋度接種2孔，同時設細胞對照和毒性對照。次日開始觀察細胞生長狀態。連續觀察5d。受試產婦40例，受試組每d1劑，煎湯分3~5次服，10d為1療程。該方藥是由鷄內金10~15份、構杞子10~15份、焦山楂5~10份、木瓜5~10份組成。將乳汁鉛水平≥0.24μmol/L和＜0.24μmol/L設為高鉛組和低鉛組。受試前後兩組間乳汁鉛水平進行比較，在3箇月齡時對研究對象用蓓蕾（Bayley）嬰兒髮育量錶進行髮育評價3箇月±1週嬰兒精神髮育指數（MDI）和心理運動髮育指數（PDI）。石墨爐原子吸收光譜法測定初乳鉛含量。結果杞金方藥製備的顆粒按不同稀釋倍數接種于生長良好的地鼠腎細胞觀察其對細胞生長的影響，經36℃連續觀察5d未見細胞固縮、腫脹、脫落等細胞毒性反應，證實杞金方藥不會引起對細胞的毒性反應。受試組和對照組服藥前後進行乳鉛水平比較，受試組乳鉛水平明顯低于對照組，兩者比較差異有顯著性。40例嬰兒3箇月齡時MDI、PDI與杞金方藥閤劑榦預的相關迴歸分析結果錶明，兩髮育指標和杞金方藥閤劑早期鉛榦預間存在有統計學意義的負相關。結論供試液按不同稀釋倍數接種于生長良好的單層地鼠腎細胞，觀察其對細胞生長的影響，單層地鼠腎細胞生長良好，與對照組比較細胞生長無差異，對地鼠腎細胞生長無不良影響。用該方藥閤劑早期榦預初乳鉛水平，對新生兒神經髮育有一定影響。
목적：아문재연구기금방약독이학급안전성적기출상，관찰이기금방약（유구기자、계내금、초산사、목과등약물조성）대지서신세포생장급조기연간예대신생인신경행위적영향，연구기대세포생장급조기연간예대신생인신경행위적발육。방법장생장상태량호적지서신세포용Versence-이매혼합액소화후제성4×105농도적세포현액접충24공세포배양판，매공1mL，37℃이양화탄배양。칭취기금방약과립제10g방입100mL배액병중，가입50mL세포배양액，121℃고압20min，4℃빙상방치24h。불단교반，병사미용해부분침정。동시장분분안동양방법처리주위독성대조조。장성단층적지서신세포추출생장액，장제비호적침제액용세포배양액1∶2~1∶32희석후가입각공，매희석도접충2공，동시설세포대조화독성대조。차일개시관찰세포생장상태。련속관찰5d。수시산부40례，수시조매d1제，전탕분3~5차복，10d위1료정。해방약시유계내금10~15빈、구기자10~15빈、초산사5~10빈、목과5~10빈조성。장유즙연수평≥0.24μmol/L화＜0.24μmol/L설위고연조화저연조。수시전후량조간유즙연수평진행비교，재3개월령시대연구대상용배뢰（Bayley）영인발육량표진행발육평개3개월±1주영인정신발육지수（MDI）화심리운동발육지수（PDI）。석묵로원자흡수광보법측정초유연함량。결과기금방약제비적과립안불동희석배수접충우생장량호적지서신세포관찰기대세포생장적영향，경36℃련속관찰5d미견세포고축、종창、탈락등세포독성반응，증실기금방약불회인기대세포적독성반응。수시조화대조조복약전후진행유연수평비교，수시조유연수평명현저우대조조，량자비교차이유현저성。40례영인3개월령시MDI、PDI여기금방약합제간예적상관회귀분석결과표명，량발육지표화기금방약합제조기연간예간존재유통계학의의적부상관。결론공시액안불동희석배수접충우생장량호적단층지서신세포，관찰기대세포생장적영향，단층지서신세포생장량호，여대조조비교세포생장무차이，대지서신세포생장무불량영향。용해방약합제조기간예초유연수평，대신생인신경발육유일정영향。
Objective The aim of study is evaluating the effect about toxicology and safety of the Qijin keli to hamster kidney cell, hamster kidney cell were cultured to determinate the significance about the Qijin keli to hamster kidney cell, evaluating the significance of foremilk lead level as an indicator for environmental lead, to investigate the relationship of early neurobehavioral development of neonates and taking Qijin herbal medication prescription about early lead intervention of foremilk exposure. Methods Determination of lead level in foremilk:altogether 40 foremilk samples, 1mL each,were collected .the same amount of violet acid was added to each sample, after foremilk was fully dissolved, 0.2mL solution was taken for determining lead level with graphite stove atom absorption spectrophotometer. The determined process strictly followed the internal quantity control of laboratory and was involved in the blind quality control of institute of Environmental Health of Chinese Academy. Results All of hamster kidney cell had no death and were analyzed. It showed that all cell behavior were normal. It had no significant difference between the test group and control group. Totally 40 neonates were involved, and the normal reference value of lead level of foremilk was 0.06 to 0.08μmol/L. The involved neonates were assigned into high-level lead group and low-level lead group. There is a relationship between the of early neurobehavioral development of neonates and taking Qijin herbal medication prescription about early lead intervention of foremilk exposure. Conclusion Qijin keli has no toxicology and side effect to hamster kidney cell various index. It has safety. Having an main effect upon neurobehavioral development and taking Qijin herbal medication prescription about early lead intervention of foremilk exposure.