中国中西医结合肾病杂志
中國中西醫結閤腎病雜誌
중국중서의결합신병잡지
CHINESE JOURNAL OF INTEGRATED TRADITIONAL AND WESTERN NEPHROLOGY
2014年
4期
306-309
,共4页
氧化低密度脂蛋白%肾小球系膜细胞%LOX-1%炎症因子
氧化低密度脂蛋白%腎小毬繫膜細胞%LOX-1%炎癥因子
양화저밀도지단백%신소구계막세포%LOX-1%염증인자
Oxidized low-density lipoprotein%Mesangial cells%LOX-1%Inflammatory cytokines
目的：应用MTT酶标分析法,检测在不同浓度、不同时长氧化低密度脂蛋白( ox-LDL)干预下的大鼠肾小球系膜细胞( RMC)的增殖情况以及不同浓度ox-LDL干预下LOX-1和TNF-α的表达,探讨其相互关系。方法：体外培养RMC,使用不同浓度ox-LDL在不同时间对RMC干预,经MTT法检测,并进行分组分析。用浓度为5μg/ml和50μg/ml的ox-LDL干预RMC 24 h,使用RT-PCR法和Western blot法检测LOX-1和TNF-α的mRNA和蛋白表达量,并进行分析比较。结果：Ox-LDL干预RMC 在时间点为24 h的结果显示,在1~30μg/ml范围内呈生长促进效应,浓度大于30μg/ml后呈生长抑制效应；干预48 h组对细胞效应趋势基本与作用24 h后一致,但生长抑制效应从30μg/ml开始；而72 h组,ox-LDL在各个浓度均显示出生长抑制效应。5μg/ml及50μg/ml浓度ox-LDL干预RMC后,均可使LOX-1和TNF-α的mRNA和蛋白表达升高,且50μg/ml较5μg/ml效果明显,结果差异具有统计学意义(P<0.05)。结论：Ox-LDL对RMC增殖影响具有浓度及时间依赖性。较低及较高浓度ox-LDL均能导致RMC中LOX-1和TNF-α的表达增高,且浓度较高时效果更明显。其机制可能为ox-LDL导致LOX-1表达增多,引起炎症因子的产生和趋化,发生细胞损伤,提示脂质代谢紊乱在肾脏疾病中的重要作用。
目的：應用MTT酶標分析法,檢測在不同濃度、不同時長氧化低密度脂蛋白( ox-LDL)榦預下的大鼠腎小毬繫膜細胞( RMC)的增殖情況以及不同濃度ox-LDL榦預下LOX-1和TNF-α的錶達,探討其相互關繫。方法：體外培養RMC,使用不同濃度ox-LDL在不同時間對RMC榦預,經MTT法檢測,併進行分組分析。用濃度為5μg/ml和50μg/ml的ox-LDL榦預RMC 24 h,使用RT-PCR法和Western blot法檢測LOX-1和TNF-α的mRNA和蛋白錶達量,併進行分析比較。結果：Ox-LDL榦預RMC 在時間點為24 h的結果顯示,在1~30μg/ml範圍內呈生長促進效應,濃度大于30μg/ml後呈生長抑製效應；榦預48 h組對細胞效應趨勢基本與作用24 h後一緻,但生長抑製效應從30μg/ml開始；而72 h組,ox-LDL在各箇濃度均顯示齣生長抑製效應。5μg/ml及50μg/ml濃度ox-LDL榦預RMC後,均可使LOX-1和TNF-α的mRNA和蛋白錶達升高,且50μg/ml較5μg/ml效果明顯,結果差異具有統計學意義(P<0.05)。結論：Ox-LDL對RMC增殖影響具有濃度及時間依賴性。較低及較高濃度ox-LDL均能導緻RMC中LOX-1和TNF-α的錶達增高,且濃度較高時效果更明顯。其機製可能為ox-LDL導緻LOX-1錶達增多,引起炎癥因子的產生和趨化,髮生細胞損傷,提示脂質代謝紊亂在腎髒疾病中的重要作用。
목적：응용MTT매표분석법,검측재불동농도、불동시장양화저밀도지단백( ox-LDL)간예하적대서신소구계막세포( RMC)적증식정황이급불동농도ox-LDL간예하LOX-1화TNF-α적표체,탐토기상호관계。방법：체외배양RMC,사용불동농도ox-LDL재불동시간대RMC간예,경MTT법검측,병진행분조분석。용농도위5μg/ml화50μg/ml적ox-LDL간예RMC 24 h,사용RT-PCR법화Western blot법검측LOX-1화TNF-α적mRNA화단백표체량,병진행분석비교。결과：Ox-LDL간예RMC 재시간점위24 h적결과현시,재1~30μg/ml범위내정생장촉진효응,농도대우30μg/ml후정생장억제효응；간예48 h조대세포효응추세기본여작용24 h후일치,단생장억제효응종30μg/ml개시；이72 h조,ox-LDL재각개농도균현시출생장억제효응。5μg/ml급50μg/ml농도ox-LDL간예RMC후,균가사LOX-1화TNF-α적mRNA화단백표체승고,차50μg/ml교5μg/ml효과명현,결과차이구유통계학의의(P<0.05)。결론：Ox-LDL대RMC증식영향구유농도급시간의뢰성。교저급교고농도ox-LDL균능도치RMC중LOX-1화TNF-α적표체증고,차농도교고시효과경명현。기궤제가능위ox-LDL도치LOX-1표체증다,인기염증인자적산생화추화,발생세포손상,제시지질대사문란재신장질병중적중요작용。
Objective:Using different concentration and duration of ox-LDL to intervene RMC,measuring the proliferation via MTT and the expression of LOX-1 and TNF-αvia rt-PCR and western blot,to explore the relationships between them. Meth-ods:Culture the RMC in vitro with different concentration and duration of ox-LDL,measure the proliferation of RMC via MTT and an-alyze the results in different groups. Use 5 μg/ml and 50 μg/ml ox-LDL to intervene RMC for 24 h individually,detect the mRNA and protein expressions of LOX-1 and TNF-αvia rt-PCR and western blot. Results:After 24 h intervention,the proliferation of RMC is promoted in the concentration of 1-30μg/ml. The proliferation is inhibited in the concentration higher than 30μg/ml. At 48 h, the trend is basically similar to 24 h,the inhibition appears at concentration of 30 μg/ml. And at 72 h,inhibitions are shown in every concentrations. After 5μg/ml and 50 μg/ml ox-LDL intervention for 24 h,the mRNA and protein expressions of LOX-1 and TNF-αare higher than the blank group. And 50μg/ml group is significantly higher than 5 μg/ml group (P<0. 05). Conclusion:The cell proliferation effect of ox-LDL on RMC is concentration-time span dependent. Ox-LDL could lead to increased expression of LOX-1 and TNF-α,and higher concentration effects more. The probable mechanism is higher concentration of ox-LDL makes higher LOX-1 expression,which leads to the production of inflammatory cytokines and chemokines,and damage the cells. It indicates the im-portant role of lipid metabolism disorder in kidney diseases.
