CAJ | 학술논문

目的了解黄蜂毒素1(MP-1)对LPS所致小鼠急性肝损伤的影响,探讨其作用机制.方法将104只BALB/c小鼠按随机数字表法分为健康对照组(8只,不作任何处理)、LPS组(48只,尾静脉注射LPS 5 mg/kg)和MP-1组(48只,尾静脉注射LPS 5 mg/kg和MP-1 3 mg/kg).后2组小鼠于注射后2、6、12、24、48、72 h处死,每组每时相点8只,采集血和肝组织标本.动态比浊法鲎试验检测2组小鼠各时相点血浆LPS水平,酶联免疫吸附测定法检测血浆TNF-α和IL-6水平,全自动生化分析仪检测血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平,实时荧光定量RT-PCR检测肝组织Toll样受体4(TLR4)、TNF-α和IL-6 mRNA表达,观察注射药物后各时相点肝组织病理变化.另取健康对照组小鼠相同样本进行以上检测.结果与健康对照组比较,LPS组小鼠血浆LPS和TNF-α水平在注射药物后2 h迅速升高,各为(18 320.50±2782.50)EU/mL和(988±130)ng/L,此后逐渐下降,72 h降至(1.80±0.80)EU/mL和(150±44)ng/L,接近健康对照组水平;IL-6、ALT和AST逐渐升高,12 h达峰值,随后下降,72 h接近健康对照组水平;LPS组小鼠肝组织TLR4、TNF-α和IL-6mRNA表达也在注射后明显增强,12～48 h时肝组织炎性反应性病理改变最为显著.与LPS组比较,MP-1组小鼠LPS、细胞因子和转氨酶水平在注射后不同程度地降低(P<0.05或P<0.01),肝组织相关基因表达受到抑制(P<0.05或P<0.01),病理改变较轻.结论 MP-1可能通过中和LPS,减少其诱导的炎性介质合成与释放,进而减轻小鼠肝组织的损伤.
목적 료해황봉독소1(MP-1)대LPS소치소서급성간손상적영향,탐토기작용궤제.방법 장104지BALB/c소서안수궤수자표법분위건강대조조(8지,불작임하처리)、LPS조(48지,미정맥주사LPS 5 mg/kg)화MP-1조(48지,미정맥주사LPS 5 mg/kg화MP-1 3 mg/kg).후2조소서우주사후2、6、12、24、48、72 h처사,매조매시상점8지,채집혈화간조직표본.동태비탁법후시험검측2조소서각시상점혈장LPS수평,매련면역흡부측정법검측혈장TNF-α화IL-6수평,전자동생화분석의검측혈청병안산전안매(ALT)화천동안산전안매(AST)수평,실시형광정량RT-PCR검측간조직Toll양수체4(TLR4)、TNF-α화IL-6 mRNA표체,관찰주사약물후각시상점간조직병리변화.령취건강대조조소서상동양본진행이상검측.결과 여건강대조조비교,LPS조소서혈장LPS화TNF-α수평재주사약물후2 h신속승고,각위(18 320.50±2782.50)EU/mL화(988±130)ng/L,차후축점하강,72 h강지(1.80±0.80)EU/mL화(150±44)ng/L,접근건강대조조수평;IL-6、ALT화AST축점승고,12 h체봉치,수후하강,72 h접근건강대조조수평;LPS조소서간조직TLR4、TNF-α화IL-6mRNA표체야재주사후명현증강,12～48 h시간조직염성반응성병리개변최위현저.여LPS조비교,MP-1조소서LPS、세포인자화전안매수평재주사후불동정도지강저(P<0.05혹P<0.01),간조직상관기인표체수도억제(P<0.05혹P<0.01),병리개변교경.결론 MP-1가능통과중화LPS,감소기유도적염성개질합성여석방,진이감경소서간조직적손상.
Objective To observe the effect of mastoparan-1 (MP-1) on lipopolysaccharide (LPS)-induced acute hepatic injury in mice and probe into its possible mechanism. Methods One hun-dred and four BALB/c mice were randomly divided into healthy control group(n=8, without treatment, HC), LPS group (n=48, with injection of LPS 5 mg/kg via tail vein), and MP-1 group (n=48, with in-jection of LPS 5 mg/kg and MP-1 3 mg/kg via tail vein). Mice in LPS group and MP-1 group were sacri-ficed at 2nd, 6th, 12th, 24th, 48th and 72nd post injection hour (PIH), 8 mice at each time point in each group. Blood samples were collected for determination of plasma levels of LPS by kinetic turbidimetric limu-lus test, TNF-α and IL-6 by ELISA, serum levels of ALT and AST by automatic biochemistry analyzer re-spectively. Hepatic tissue samples were collected for determination of TLR4, TNF-α and IL-6 mRNA by re-al-time fluorescent quantitation reverse transcription polymerase chain reaction, along with the observation of pathological changes in hepatic tissue at each time point. Above-mentioned examinations were also performed in HC group. Results Compared with those of HC group, plasma levels of LPS and TNF-α in LPS group significantly increased at 2nd PIH(18 320.50±2782.50 EU/mL and 988±130 ng/L, respectively), then decreased gradually to 1.80±0.80 EU/mL and 150±44 ng/L at 72nd PIH, which was close to those of HC group. The values of IL-6, ALT and AST peaked at 12th PIH, which declined to the levels close to those of HC group at 72nd PIH. Meanwhile, the expressions of TLR4, TNF-α and IL-6 mRNA in liver were remark-ably up-regulated after injection, and the pathological changes in hepatic tissue pronounced significantly at 12nd, 24th and 48th PIH. Compared with those of LPS group, the levels of LPS, cytokines, ALT and AST de-creased in MP-1 group in different degrees after injection (P <0.05 or P <0. 01), genes expression(P < 0.05 or P <0.01) and pathological changes was respectively suppressed and alleviated in hepatic tissue. Conclusions MP-1 can alleviate LPS-induced acute hepatic injury in mice, which may be associated with its neutralization of LPS and attenuation of synthesis and release of inflammatory mediators.