中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2009年
3期
175-178
,共4页
郑贵军%武子霞%李银平%姚咏明
鄭貴軍%武子霞%李銀平%姚詠明
정귀군%무자하%리은평%요영명
脂多糖%主动脉内皮细胞%内皮细胞蛋白C受体%蛋白酶活化受体1%血必净注射液%大鼠
脂多糖%主動脈內皮細胞%內皮細胞蛋白C受體%蛋白酶活化受體1%血必淨註射液%大鼠
지다당%주동맥내피세포%내피세포단백C수체%단백매활화수체1%혈필정주사액%대서
lipopolysaccharide%aortic endothelial cell%endothelial protein C receptor%protease activated receptor 1%Xuebijing injection%rat
目的 观察脂多糖(LPS)诱导的大鼠主动脉内皮细胞蛋白C受体(EPCR)和蛋白酶活化受体1(PAR1)的表达,并探讨血必净注射液对其的干预作用.方法 采用组织贴块法培养Wistar大鼠主动脉内皮细胞,培养1周后用流式细胞仪鉴定内皮细胞纯度.按1∶3传代至3～4代时,随机将细胞分为空白对照组、LPS刺激组(1 mg/L)、血必净干预组(LPS 1 mg/L,血必净注射液10 g/L),活化蛋白C(APC)干预组(LPS 1 mg/L,APC 0.1 mg/L).分别在12、24、48、72 h采用逆转录-聚合酶链反应(RT-PCR)测定内皮细胞EPCR和PAR1的mRNA表达;用流式细胞仪检测EPCR和PAR1的蛋白表达.结果 12 h时各组间EPCR和PAR1蛋白表达水平比较差异均无统计学意义(P均>0.05);LPS刺激组EPCR和PAR1 mRNA表达则均降低,APC和血必净干预后两值均升高(P<0.05或P<0.01).24～72 h LPS刺激组EPCR的mRNA和蛋白表达水平均明显低于空白对照组(P<0.05或P<0.01),EPCR的表达随LPS作用时间延长而减少,呈明显的负相关;而血必净干预组EPCR表达明显高于LPS刺激组(P均<0.01).LPS刺激组PAR1的mRNA和蛋白表达水平较空白对照组有所降低(P<0.05或P<0.01),用血必净干预后,PAR1的mRNA和蛋白表达水平有所增加(P均<0.01).与APC干预组比较,血必净干预组EPCR和PAR1的蛋白表达增加更为明显.结论 血必净注射液能从基因和蛋白水平增加由LPS诱导大鼠主动脉EPCR和PAR1的表达作用,可能与其保护内皮细胞的功能抑制其凋亡有关.
目的 觀察脂多糖(LPS)誘導的大鼠主動脈內皮細胞蛋白C受體(EPCR)和蛋白酶活化受體1(PAR1)的錶達,併探討血必淨註射液對其的榦預作用.方法 採用組織貼塊法培養Wistar大鼠主動脈內皮細胞,培養1週後用流式細胞儀鑒定內皮細胞純度.按1∶3傳代至3～4代時,隨機將細胞分為空白對照組、LPS刺激組(1 mg/L)、血必淨榦預組(LPS 1 mg/L,血必淨註射液10 g/L),活化蛋白C(APC)榦預組(LPS 1 mg/L,APC 0.1 mg/L).分彆在12、24、48、72 h採用逆轉錄-聚閤酶鏈反應(RT-PCR)測定內皮細胞EPCR和PAR1的mRNA錶達;用流式細胞儀檢測EPCR和PAR1的蛋白錶達.結果 12 h時各組間EPCR和PAR1蛋白錶達水平比較差異均無統計學意義(P均>0.05);LPS刺激組EPCR和PAR1 mRNA錶達則均降低,APC和血必淨榦預後兩值均升高(P<0.05或P<0.01).24～72 h LPS刺激組EPCR的mRNA和蛋白錶達水平均明顯低于空白對照組(P<0.05或P<0.01),EPCR的錶達隨LPS作用時間延長而減少,呈明顯的負相關;而血必淨榦預組EPCR錶達明顯高于LPS刺激組(P均<0.01).LPS刺激組PAR1的mRNA和蛋白錶達水平較空白對照組有所降低(P<0.05或P<0.01),用血必淨榦預後,PAR1的mRNA和蛋白錶達水平有所增加(P均<0.01).與APC榦預組比較,血必淨榦預組EPCR和PAR1的蛋白錶達增加更為明顯.結論 血必淨註射液能從基因和蛋白水平增加由LPS誘導大鼠主動脈EPCR和PAR1的錶達作用,可能與其保護內皮細胞的功能抑製其凋亡有關.
목적 관찰지다당(LPS)유도적대서주동맥내피세포단백C수체(EPCR)화단백매활화수체1(PAR1)적표체,병탐토혈필정주사액대기적간예작용.방법 채용조직첩괴법배양Wistar대서주동맥내피세포,배양1주후용류식세포의감정내피세포순도.안1∶3전대지3～4대시,수궤장세포분위공백대조조、LPS자격조(1 mg/L)、혈필정간예조(LPS 1 mg/L,혈필정주사액10 g/L),활화단백C(APC)간예조(LPS 1 mg/L,APC 0.1 mg/L).분별재12、24、48、72 h채용역전록-취합매련반응(RT-PCR)측정내피세포EPCR화PAR1적mRNA표체;용류식세포의검측EPCR화PAR1적단백표체.결과 12 h시각조간EPCR화PAR1단백표체수평비교차이균무통계학의의(P균>0.05);LPS자격조EPCR화PAR1 mRNA표체칙균강저,APC화혈필정간예후량치균승고(P<0.05혹P<0.01).24～72 h LPS자격조EPCR적mRNA화단백표체수평균명현저우공백대조조(P<0.05혹P<0.01),EPCR적표체수LPS작용시간연장이감소,정명현적부상관;이혈필정간예조EPCR표체명현고우LPS자격조(P균<0.01).LPS자격조PAR1적mRNA화단백표체수평교공백대조조유소강저(P<0.05혹P<0.01),용혈필정간예후,PAR1적mRNA화단백표체수평유소증가(P균<0.01).여APC간예조비교,혈필정간예조EPCR화PAR1적단백표체증가경위명현.결론 혈필정주사액능종기인화단백수평증가유LPS유도대서주동맥EPCR화PAR1적표체작용,가능여기보호내피세포적공능억제기조망유관.
Objective To investigate the effect of Xuebijing injection (血必净注射液) on expression of endothelial protein C receptor (EPCR) and protease activated receptor 1 (PAR1) mRNA and protein in rat aortic endothelial cells (RAECs) after lipopolysaccharide (LPS) challenge. Methods RAECs were cultured for one week, and the purity was determined with flow cytometry. ARECs were randomly divided into four groups: control group, LPS stimulation group (1 mg/L LPS), Xuebijing injection treatment group (LPS: 1 mg/L, Xuebijing injection: 10 g/L) and activated protein C (APC) treatment group (LPS: 1 mg/L, APC: 0.1 mg/L). The RAECs were collected at 12, 24, 48, and 72 hours after being stimulated by LPS. EPCR and PAR1 mRNA of RAECs were assessed by reverse transcription-polymerase chain reaction (RT-PCR). EPCR and PAR1 protein were assessed by flow cytometry. Results At 12 hours, EPCR and PAR1 protein expressions were not significantly different among groups (all P>0.05). The level of EPCR and PAR1 mRNA were decreased in LPS stimulation group, but they were elevated in both APC and Xuebijing injection treatment groups (P<0.05 or P<0.01). From 24 to 72 hours, compared to control group, the levels of EPCR mRNA and protein expression were significantly decreased after LPS stimulation (P<0.05 or P<0.01). The levels of EPCR expression were decreased and negatively correlated with the time of LPS treatment. Also, compared to LPS stimulation group, treatment with Xuebijing markedly elevated the levels of EPCR (P<0.01). The levels of PAR1 expression were significantly decreased by LPS stimulation compared with those of control group (P<0.05 or P<0.01). After the treatment with Xuebijng, the expression of PAR1 was gradually increased (all P<0.01). Compared with APC treatment group, Xuebijing could increase the PAR1 expression better. Conclusion Xuebijing could raise EPCR and PAR1 mRNA and protein expression of RAECs after LPS challenge, and it may be related to its protection of endothelial cell from undergoing apoptosis.
