CAJ | 학술논문

目的：探讨前列地尔对早期百草枯中毒大鼠急性肝损伤的保护作用及其机制。方法72只大鼠随机分为对照组、染毒组和治疗组，每组同时分为染毒后1、3和7d共3个时间点，每组每个时间点8只大鼠。染毒组和治疗组大鼠均腹腔内注射20％百草枯浓缩液30 mg/kg建立百草枯中毒模型，对照组大鼠仅腹腔内注射30 mg/kg 0.9％氯化钠溶液。1 d后治疗组大鼠腹腔内注射前列地尔注射液1μg/kg，1次/d，直至处死前；其余两组大鼠腹腔内注射等体积无菌生理盐水。在相应时间点处死大鼠后收集肝组织及血清，肝组织HE染色后光学显微镜下观察；ELISA法检测血清细胞因子肿瘤坏死因子-α（ TNF-α）和白细胞介素-1β（ IL-1β）表达；RT-PCR法检测诱导型一氧化氮合酶（ iNOS）和p53的表达；收集染毒后3 d肝组织用底物显色法检测Caspase-3、8、9、12活性。结果经药物处理后，染毒组出现弥漫性的大片坏死，炎性细胞浸润，肝细胞再生不明显；而治疗组肝小叶结构尚存，偶可见肝细胞点状坏死，有少量充血及炎性细胞浸润。各时间点中，染毒组的血清TNF-α和IL-1β水平较对照组显著升高（P＜0.01），而治疗组则较染毒组显著降低（P＜0.01）。染毒后3 d染毒组的Caspase-3、8、9、12活性显著高于对照组（P＜0.01），而治疗组则较染毒组显著降低（P＜0.01）。染毒组iNOS和p53基因表达较对照组显著升高（P＜0.01），而治疗组明显较染毒组降低（P＜0.01）。结论前列地尔能明显降低早期百草枯中毒大鼠炎性因子水平，抑制Caspase-3、8、9、12活性及iNOS、p53基因表达，对早期百草枯中毒大鼠具有肝保护作用。
목적：탐토전렬지이대조기백초고중독대서급성간손상적보호작용급기궤제。방법72지대서수궤분위대조조、염독조화치료조，매조동시분위염독후1、3화7d공3개시간점，매조매개시간점8지대서。염독조화치료조대서균복강내주사20％백초고농축액30 mg/kg건립백초고중독모형，대조조대서부복강내주사30 mg/kg 0.9％록화납용액。1 d후치료조대서복강내주사전렬지이주사액1μg/kg，1차/d，직지처사전；기여량조대서복강내주사등체적무균생리염수。재상응시간점처사대서후수집간조직급혈청，간조직HE염색후광학현미경하관찰；ELISA법검측혈청세포인자종류배사인자-α（ TNF-α）화백세포개소-1β（ IL-1β）표체；RT-PCR법검측유도형일양화담합매（ iNOS）화p53적표체；수집염독후3 d간조직용저물현색법검측Caspase-3、8、9、12활성。결과경약물처리후，염독조출현미만성적대편배사，염성세포침윤，간세포재생불명현；이치료조간소협결구상존，우가견간세포점상배사，유소량충혈급염성세포침윤。각시간점중，염독조적혈청TNF-α화IL-1β수평교대조조현저승고（P＜0.01），이치료조칙교염독조현저강저（P＜0.01）。염독후3 d염독조적Caspase-3、8、9、12활성현저고우대조조（P＜0.01），이치료조칙교염독조현저강저（P＜0.01）。염독조iNOS화p53기인표체교대조조현저승고（P＜0.01），이치료조명현교염독조강저（P＜0.01）。결론전렬지이능명현강저조기백초고중독대서염성인자수평，억제Caspase-3、8、9、12활성급iNOS、p53기인표체，대조기백초고중독대서구유간보호작용。
Objective To observe the liver protection of alprostadil against paraquat -induced early acute liver injury in rats and its mechanism .Methods 72 rats were divided into control group , exposure group and treatment group , among which each 8 rats were subasigned to different durations of exposure by 1, 3 and 7 days.The rats in the exposure and treatment groups received intraperitoneal injections of 20%paraquat (30 mg/kg) to establish the paraquat -poisoning model, while the rats in control group were treated with sterile 0.9%sodium chloride solution (30 mg/kg) only.The rats in treatment group received intraperitoneal injection of alprostadil (1μg/kg) evreyday until excution , while replaced ster-ile saline were given to the other 2 groups .The rats were sacrificed in the corresponding time points and their sera and liv-er tissues were collected .Liver tissues were fixed and stained with hematoxylin and eosin ( HE) for optical microscopic ex-amination.The serum cytokine expression was assessed by ELISA .The inducible nitric oxide synthase ( iNOS) and p53 genes were assessed by RT-PCR, while the liver Caspase-3, -8, -9, and -12 activities were measured by chromo-genic substrate method 3 days after exposure .Results Diffuse large areas of necrosis , inflammatory cell infiltration and impaired hepatocell regeneration were observed in exposure group ;while spared lobular architecture , spotted necrosis , low degreed of congestion and inflammatory cell infiltration were observed in treatment group .The serum TNF-αand IL-1βlevels were significantly higher in exposure group than control group (P<0.01), which was significantly reversed in treat-ment group (P<0.01).Liver Caspase-3, -8, -9 and -12 activities were significantly higher in exposure group than control group (P<0.01), which were also singificantly reversed in treatment group (P<0.01);so was the expression of iNOS and p53 ( P<0.01) .Conclusion Alprostadil can significantly reduce the inflammatory cytokine levels , inhibit the Caspase-3, -8, -9 and -12 activities, and down-regulate the expression of iNOS and p 53 in early paraquat -poi-soning rats, thus provides protective effects against liver injury .