【摘要】 目的 觀察百草枯中毒大鼠腎組織中血紅素氧合酶1(HO1)的表達,探討其病理生理機制。 方法 SD大鼠126只隨機分為空白對照組、中毒組和褪黑素組,各42只。中毒組、褪黑素組予以百草枯(25 mg/kg)腹腔注射染毒,對照組予以等量生理鹽水腹腔注射,15 min后褪黑素組予以褪黑素(10 mg/kg)腹腔注射,對照組、中毒組予以等量生理鹽水腹腔注射。于1、3、6、12 h,1、2、3、5 d時各組隨機取6只處死。蘇木精〖CD3/5〗伊紅（HE）染色觀察各組腎組織病理學變化,采用免疫組識化學和RTPCR觀察腎組織HO1蛋白和mRNA表達。 結果 ①與對照組相比,中毒組染毒后3 h即可見充血、水腫及空泡變性等病理變化,1 d達頂峰,病理損傷評分3.30±0.31(Plt;0.05),其后緩解趨勢不明顯；而褪黑素組病理變化明顯減輕且緩解趨勢明顯,1 d時病理損傷評分2.70±0.26,與中毒組相比差異有統計學意義(Plt;0.05)。②與對照組相比,中毒組染毒3 h在皮質部腎小管上皮細胞的細胞膜及細胞漿HO1呈陽性表達,免疫組識化學評分(IHS)3.33±1.75,HO1 mRNA表達增強,與對照組相比差異有統計學意義(Plt;0.05),1 d達頂峰,HIS為7.00±2.00,之后減弱,5 d仍有陽性表達,但與對照組相比差異無統計學意義(Pgt;0.05)；褪黑素組HO1表達較中毒組明顯增強,IHS評分6 h~3 d差異具有統計學意義(Plt;0.05),5 d不再有統計學意義(Pgt;005)。 結論 HO1在百草枯中毒大鼠腎組織中呈高表達,褪黑素能明顯改善百草枯中毒腎臟病理損傷,增強HO1表達可能是其作用途徑之一,而氧化損傷可能是百草枯中毒腎損傷病理生理機制之一。【Abstract】 Objective To investigate the expression of heme oxygenase1 (HO1) in paraquartinduced renal injury in rats and the protective effects of melatonin, and explore possible mechanism of paraquartinduced renal injury. Methods One hundred and twentysix adult healthy SpragneDawley rats were randomly divided into three groups and 42 in each group: control group (A), paraquart group (B), and melatonin group (C). The rats in group B and group C were treated with paraquart (25 mg/kg) intraperitoneally, the rats in group A were treated with the same dose of normal saline. In 15 minutes, the rats in group C were given melatonin intraperitoneally at a dose of 10 mg/(kg·d) and the rats in group A and B were treated intraperitoneally with the same dose of normal saline. Six rats in each group were randomly sacrificed at one, three, 12 hours and one, tou, three, five days respectively. Renal histopathological changes were observed under light microscope by HE staining. The protein and mRNA expressions of HO1 were evaluated by immunohistochemical staining and RTPCR respectively. Results ①In group B, there were obvious lesions in the renal tubule of cortical part, including edema, congestion and vacuolar degeneration. These pathologic changes gradually reached the peak on the first day and did not relieved till the end of this study, the pathologic injury score was 3.30±0.31, and there was a statistical significance between group B and group A (Plt;0.05). The aforementioned pathological lesion was more palliative in group C, the pathologic injury score was 2.70±0.26 at the first day; Compared with group B, there was a statistical significance. ②In group A, there was no or weak expression of HO1 and HO1 mRNA. At the third hour, the expression of HO1 in group B was observed in the membrane and cytoplasm of renal tubular epithelial cell of cortical part. Immunohistochemistry score (IHS) was 3.33±1.75 and the expression of HO1 mRNA increased, there was a statistical significance between group B and group A (Plt;0.05). It reached the peak on the first day, IHS was 7.00±2.00, but there was no statistical difference between group B and group A on the fifth day (Pgt;0.05); Compared with group B, the expression in group C was enhanced obviously, IHS were higher obviously on the six hour till to the third day (Plt;0.05), but there were no statistical differences on the fifth day (Pgt;0.05). Conclusion The expression of HO1 in the paraquartdamaged kidney increases and melatonin surely has an protective effect by increasing the expression of HO1, which suggests that oxidative injury might be the main mechanism of paraquartinduced renal injury.
Objective To investigate the effects and mechanisms of hydroxychloroquine sulfate (HCQ) on pulmonary fibrosis through the PI3K/AKt/mTOR signalling pathway. Methods Paraquat intraperitoneal injection was used to establish a mouse model of pulmonary fibrosis. Thirty-six SPF C57BL/6J female mice were randomly divided into a blank group, a paraquat group (20 mg/kg) and a HCQ intervention group. The HCQ intervention group was divided into two subgroups (10 mg/kg and 30 mg/kg) according to different doses. The general condition and body weight changes of mice were observed. twenty-one days later, lung tissues were stained with hematoxylin-eosin and Masson’s pathological staining, and the content of inflammatory factors (IL-1β, IL-6, TNF-α) and hydroxyproline (HYP) were detected by ELISA. Alpha-smooth muscle actin (α-SMA), E-cadherin (E-cad), the expression levels of PI3K/Akt/mTOR pathway-related proteins, phosphatidylinositol 3 kinase (PI3K) and protein kinase B (AKt), and mammalian target of rapamycin (mTOR) were detected by Western blot. The gene expression levels of α-SMA and E-cad were detected by q-PCR. Results Compared with the blank group, the mice in the paraquat group had lower body weight, worse general condition, higher serum levels of inflammatory factors, increased lung structure destruction and collagen deposition, significantly increased HYP content, and higher expression level of PI3K/AKt/mTOR signaling pathway related proteins (all P<0.05). The expression levels of E-cad protein and gene decreased, α-SMA protein and gene increased (all P<0.05). While the HCQ intervention group improved the degree of pulmonary fibrosis in different degrees, and the relevant indexes of PI3K/AKt/mTOR signaling pathway decreased compared with the paraquat group (all P<0.05). Conclusion HCQ can ameliorate paraquat-induced pulmonary fibrosis by inhibiting the PI3K/AKt/mTOR signaling pathway.
目的 探討急性百草枯(PQ)中毒鼠肺組織病理損傷和肺組織血紅素氧合酶-1（HO-1）的表達及三七總皂甙（PNS）的保護作用。 方法 150只SD雄性鼠分為正常對照組（C組）30只、PQ中毒組（PQ組）60只及PNS組60只。PQ組和PNS組一次性灌胃PQ 25 mg/kg染毒，C組給予等體積生理鹽水灌胃。其中PNS組于染毒前15 min以PNS 50 mg/kg陰莖靜脈注射保護，以后1次/d給藥直至處死前；PQ組、C組分別在同時間點給予等體積生理鹽水。觀察各組大鼠在中毒后6、12 h，1、3、5、7 d肺組織病理改變，采用蛋白質印跡法分析肺組織HO-1蛋白表達和反轉錄-聚合酶鏈反應方法測定鼠肺組織HO-1 mRNA的表達。 結果 C組HO-1蛋白和HO-1 mRNA絕大多數標本有弱表達，個別標本不表達；與C組相比PQ組及PNS組HO-1蛋白和HO-1 mRNA表達增強，差異有統計學意義（P＜0.05）；PQ組HO-1蛋白和HO-1 mRNA的表達在1 d達高峰之后下降，第3天基本恢復到C組水平；PNS組與PQ組相似，但在6 h、12 h、1 d及3 d高于PQ組，差異有統計學意義（P＜0.05），至第5天和第7天二者相比差異無統計學意義（P＜0.05）。PQ組肺組織病理損傷評分在6、12 h，1、3 、5、7 d各亞組均高于PNS相應組，差異有統計學意義（P＜0.05），C組肺組織病理大致正常，與PQ組及PNS組相比，差異有統計學意義（P＜0.001）。 結論 HO-1參與PQ中毒所致急性肺損傷，PNS對PQ中毒所致急性肺損傷有保護作用。
ObjectiveTo establish paraquat (PQ)-induced acute respiratory distress syndrome (ARDS) mice model via gavage, in order to simulate oral adminitration in clinical situations.MethodsSeventy-eight 6-8-week-old, specific pathogen free female C57 mice were chosen in this study. The mice were randomly divided into the control group (n=6) and the PQ model group(n=36); the mice in the latter group were randomly divided into 6 poisoning model subgroups further, with 6 mice in each, to find out the suitable concentration of PQ to establish stable ARDS model. The mice in the control group were given phosphatebuffer saline (PBS) by gavage, 200 μL per mouse; while the mice in the 6 poisoning model subgroups were given PQ with varies doses of 3, 10, 30, 100, 150, 300 mg/kg respectively by gavage. The clinical manifestations were observed for 7 days, and the ratio of lung wet/dry (W/D) was measured. After the suitable concentration of PQ for stable ARDS mice model was found, the other 36 mice were randomly divided into the controlgroup and the poisoning model group, both were divided into 4 subgroups, according to different observation point in time (1 day and 2, 3, 4 days after PQ gavage). The mice in the 4 control subgroups (n=3) were given PBS by gavage, 200 μL per mouse; while the mice in the 4 poisoning model subgroups (n=6) were given PQ with the suitable concentration for ARDS mice model by gavage. Pathological manifestations by Haematoxylin-Eosin staining and lung injury score were observed and analyzed.ResultsThe mice began to die at the PQ dosage of 150 mg/kg; while the death rate was stable at 300 mg/kg. On the 2nd and 4th day after PQ gavage, lung W/D was 5.335, 6.113, and 5.525, and 6.403, respectively in the mice in 150 and 300 mg/kg subgroup, which differed much from those in the control group (P<0.001). Congestion, edema, hemorrhage, alveolar structure damage, inflammation cells infiltration of lung tissue were observed, and lung injury score increased.ConclusionPQ-induced ARDS mice model by gavage is established successfully.
【摘要】 目的 探討口服百草枯中毒患者口腔護理的方法及早期護理的臨床意義。 方法 2009年1月-2010年3月，采用半隨機方法將62例白草枯中毒患者按中毒時間的長短分A組（中毒時間lt;3 d，n=32）、B組（中毒時間gt;3 d，n=3），比較兩組患者口腔潰瘍的治愈率、并發癥發生狀況，分析早期口腔護理的必要性。 結果 A組患者百草枯所致的口腔黏膜損害明顯減輕，并發癥發生率降低，為改善預后提供了條件，顯示了早期加強口腔護理的成效。 結論 重視百草枯早期口腔護理，能夠減輕口腔糜爛潰瘍痛苦，減少并發癥，提高患者生活質量。【Abstract】 Objective To investigate the clinical significance of early oral care for paraquat-poisoned patients. Methods A quasi-randomized controlled trial was used. A total of 62 paraquat-poisoned patients (from January 2009 to March 2010) were divided into experimental group and control group in order to compare the healing rate of oral ulcer, complications and the necessity of early oral care between the two groups. Results The oral mucosa lesions in experimental group obviously alleviated and the complications decreased. The effective early oral care provided the very favorable conditions for better prognosis. Conclusion The early oral care for paraquat-poisoned patients could relieve the pain of oral ulcer, reduce the complications and improve patient′s life quality.
目的:探討貫葉連翹提取物對百草枯誘導的大鼠肺纖維化模型的干預作用及機制。方法:20只Sprague-dawly大鼠隨機等分為4組:百草枯組,貫葉連翹提取物組,百草枯+貫葉連翹提取物組和對照組。百草枯液按80mg/kg一次性灌胃，貫葉連翹提取物按400mg/kg灌胃，連用3d,對照組僅用生理鹽水。于21d處死后取肺臟行HE染色鑒定,并用堿水解法測羥脯氨酸含量,同時以八木國夫熒光法測定肺組織脂質過氧化產物,鹽酸羥胺法測定組織總超氧化物歧化酶的酶活力。結果:百草枯染毒動物可致肺纖維化,羥脯氨酸和丙二醛含量明顯增高。百草枯+貫葉連翹提取物組與百草枯組比較,第21天時肺纖維化減輕,羥脯氨酸和丙二醛含量減少(Plt;0.05),與對照組相比總超氧化物歧化酶無明顯變化(Pgt;0.05)。結論:貫葉連翹提取物對百草枯誘導的大鼠肺纖維化模型有抑制作用,其機制可能與抑制脂質過氧化有關。