中国医药科学
中國醫藥科學
중국의약과학
CHINA MEDICINE AND PHARMACY
2014年
20期
7-12,26
,共7页
蔡宏瑜%杨光辉%万强%张娟%傅志慧%高艳霞
蔡宏瑜%楊光輝%萬彊%張娟%傅誌慧%高豔霞
채굉유%양광휘%만강%장연%부지혜%고염하
黄芪山甲方%肾间质纤维化%TGF-β1%FN
黃芪山甲方%腎間質纖維化%TGF-β1%FN
황기산갑방%신간질섬유화%TGF-β1%FN
Huangqishanjia%Renal interstitial fibrosis%TGF-β1%FN
目的：动态观察黄芪山甲方（HQSJ）对大鼠肾间质纤维化（RIF）多靶点的治疗作用，探讨HQSJ方在不同阶段的可能作用机制。方法实验采用UUO大鼠模型，将雄性SD大鼠54只随机分成3组：假手术组（Sham）、模型组（UUO）、治疗组（HQSJ）。术后14、28、56d分别处死6只大鼠，观察肾功能变化；对肾小管基底膜（TBM）、肾小管萎缩、炎细胞浸润及间质纤维化情况进行半定量评分；采用免疫组织化学法及RT-PCR法检测肾组织TGF-β1、FN蛋白及mRNA的表达。结果（1）肾功能方面：与Sham组比较，UUO及HQSJ组大鼠BUN、SCr值在14～28d进行性升高，在28～56d相对稳定。HQSJ组大鼠BUN、SCr值在同一时相均较UUO组低（P＜0.05）。（2）病理改变方面：与Sham组比较，UUO及HQSJ组大鼠炎细胞浸润与肾间质慢性病变程度均有增高趋势，同一时相比较差异均有统计学意义（P＜0.05）。UUO与HQSJ组比较，炎细胞浸润程度在14、28d有统计学意义（P＜0.05），56d无统计学意义（P＞0.05）；肾间质慢性病变程度14、28、56d均有统计学意义（P＜0.05）。（3）TGF-β1、FN表达：与Sham组比较，UUO及HQSJ组TGF-β1、FN表达均增高，在同一时相比较均有统计学意义（P＜0.05）；与UUO组比较，HQSJ组TGF-β1、FN在14、28、56d的表达均不同程度下降（P＜0.05）；同一时相下降幅度比较：免疫组化示TGF-β1、FN蛋白阳性率分别下降64％、47％、7％及82％、72％、70％；RT-PCR法检测TGF-β1、FN的mRNA分别下降72％、57％、27％及83％、72％、68％。结论 HQSJ方抗肾间质纤维化的多靶点作用，可能与减少炎性细胞、抑制TGF-β1分泌，减轻肾脏固有细胞的活化及转分化，从而减少细胞外基质的沉积有关。其作用从病变早期开始，在14～28d主要是减少炎性细胞浸润和降低TGF-β1表达，对FN的抑制作用贯穿始终。
目的：動態觀察黃芪山甲方（HQSJ）對大鼠腎間質纖維化（RIF）多靶點的治療作用，探討HQSJ方在不同階段的可能作用機製。方法實驗採用UUO大鼠模型，將雄性SD大鼠54隻隨機分成3組：假手術組（Sham）、模型組（UUO）、治療組（HQSJ）。術後14、28、56d分彆處死6隻大鼠，觀察腎功能變化；對腎小管基底膜（TBM）、腎小管萎縮、炎細胞浸潤及間質纖維化情況進行半定量評分；採用免疫組織化學法及RT-PCR法檢測腎組織TGF-β1、FN蛋白及mRNA的錶達。結果（1）腎功能方麵：與Sham組比較，UUO及HQSJ組大鼠BUN、SCr值在14～28d進行性升高，在28～56d相對穩定。HQSJ組大鼠BUN、SCr值在同一時相均較UUO組低（P＜0.05）。（2）病理改變方麵：與Sham組比較，UUO及HQSJ組大鼠炎細胞浸潤與腎間質慢性病變程度均有增高趨勢，同一時相比較差異均有統計學意義（P＜0.05）。UUO與HQSJ組比較，炎細胞浸潤程度在14、28d有統計學意義（P＜0.05），56d無統計學意義（P＞0.05）；腎間質慢性病變程度14、28、56d均有統計學意義（P＜0.05）。（3）TGF-β1、FN錶達：與Sham組比較，UUO及HQSJ組TGF-β1、FN錶達均增高，在同一時相比較均有統計學意義（P＜0.05）；與UUO組比較，HQSJ組TGF-β1、FN在14、28、56d的錶達均不同程度下降（P＜0.05）；同一時相下降幅度比較：免疫組化示TGF-β1、FN蛋白暘性率分彆下降64％、47％、7％及82％、72％、70％；RT-PCR法檢測TGF-β1、FN的mRNA分彆下降72％、57％、27％及83％、72％、68％。結論 HQSJ方抗腎間質纖維化的多靶點作用，可能與減少炎性細胞、抑製TGF-β1分泌，減輕腎髒固有細胞的活化及轉分化，從而減少細胞外基質的沉積有關。其作用從病變早期開始，在14～28d主要是減少炎性細胞浸潤和降低TGF-β1錶達，對FN的抑製作用貫穿始終。
목적：동태관찰황기산갑방（HQSJ）대대서신간질섬유화（RIF）다파점적치료작용，탐토HQSJ방재불동계단적가능작용궤제。방법실험채용UUO대서모형，장웅성SD대서54지수궤분성3조：가수술조（Sham）、모형조（UUO）、치료조（HQSJ）。술후14、28、56d분별처사6지대서，관찰신공능변화；대신소관기저막（TBM）、신소관위축、염세포침윤급간질섬유화정황진행반정량평분；채용면역조직화학법급RT-PCR법검측신조직TGF-β1、FN단백급mRNA적표체。결과（1）신공능방면：여Sham조비교，UUO급HQSJ조대서BUN、SCr치재14～28d진행성승고，재28～56d상대은정。HQSJ조대서BUN、SCr치재동일시상균교UUO조저（P＜0.05）。（2）병리개변방면：여Sham조비교，UUO급HQSJ조대서염세포침윤여신간질만성병변정도균유증고추세，동일시상비교차이균유통계학의의（P＜0.05）。UUO여HQSJ조비교，염세포침윤정도재14、28d유통계학의의（P＜0.05），56d무통계학의의（P＞0.05）；신간질만성병변정도14、28、56d균유통계학의의（P＜0.05）。（3）TGF-β1、FN표체：여Sham조비교，UUO급HQSJ조TGF-β1、FN표체균증고，재동일시상비교균유통계학의의（P＜0.05）；여UUO조비교，HQSJ조TGF-β1、FN재14、28、56d적표체균불동정도하강（P＜0.05）；동일시상하강폭도비교：면역조화시TGF-β1、FN단백양성솔분별하강64％、47％、7％급82％、72％、70％；RT-PCR법검측TGF-β1、FN적mRNA분별하강72％、57％、27％급83％、72％、68％。결론 HQSJ방항신간질섬유화적다파점작용，가능여감소염성세포、억제TGF-β1분비，감경신장고유세포적활화급전분화，종이감소세포외기질적침적유관。기작용종병변조기개시，재14～28d주요시감소염성세포침윤화강저TGF-β1표체，대FN적억제작용관천시종。
Objective To observe the multi-target therapeutic effect of HQSJ for RIF of mice and discuss possible action mechanism of HQSJ in different stages. Methods UUO mice mode was adopted and 54 male SD mice were randomly divided into 3 groups as fake operation group (Sham), model group (UUO) and treatment group (HQSJ). 6 mice were killed separately on 14d, 28d and 56d after operation and their kidney function changes were observed;TBM, renal tubular atrophy, inflammatory cell infiltration and RIF were given semi-quantitative scores;TGF-β1, FN protein and mRNA expressions of renal tissues were detected by immunohistochemical method and RT-PCR method. Results (1) On kidney function, compared with Sham group, BUN and SCr values of UUO and HQSJ group were developed during 14-28d and stayed relatively steady during 28-56d. BUN and SCr values of HQSJ group were both lower than UUO group in the same time phase (P < 0.05). (2) On pathological chages, compared with Sham group, inflammatory cell infiltration and renal interstitial chronic lesion degrees of mice of UUO and HQSJ group both had a increasing trend in the same time phase (P<0.05). There were significant differences between inflammatory cell infiltration of UUO group and HQSJ group on 14 and 28d (P < 0.05) but no significant difference on 56d (P > 0.05); there were significant differences on renal interstitial chronic lesion degrees on 14, 28 and 56d (P<0.05). (3) On TGF-β1, FN expressions:compared with Sham group, TGF-β1 and FN expressions of UUO and HQSJ group both rose in the same time phase (P<0.05); compared with UUO group, TGF-β1 and FN expressions on 14, 28 and 56d of HQSJ group all declined to different extents (P<0.05);decline degree of the same time phase comparison;immunohistochemical method showed that positive rates of TGF-β1 and FN protein declined separately by 64%, 47%, 7% and 82%, 72%, 70%; RT-PCR method showed that mRNA of TGF-β1 and FN declined separately by 72%, 57%, 27%%and 83%, 72%, 68%. Conclusion Multi-target therapeutic effect of HQSJ for RIF may related with decrease of ECM deposition caused by decrease of inflammatory cells, inhibition of TGF-β1 secretion, alleviation of excitation and transdifferentiation of renal inherent cells. Its action started from early lesion, mainly reducing inflammatory cell infiltration and decreasing TGF-β1 expression, and inhibition to FN throughout the whole process.
