目的:研究长链非编码RNA HOTAIR调控糖皮质激素受体的表达对急性淋巴细胞白血病细胞增殖及凋亡的作用.方法:采用RT-qPCR法检测人正常骨髓基质细胞系HS-5及急性淋巴细胞白血病细胞系MOLT-4、CCRF-CEM和CEM-C1中HOTAIR和糖皮质激素受体的表达.用siRNA沉默CEM-C1细胞中HOTAIR的表达,CCK-8法检测si-HOTAIR对CEM-C1细胞活力的影响;BrdU法检测si-HOTAIR对CEM-C1细胞增殖的影响以及对地塞米松抑制CEM-C1细胞增殖的增效作用;Hoechst 33342染色法和caspase 3/7活性检测法研究si-HOTAIR对CEM-C1细胞凋亡的影响;并采用Western blot法检测糖皮质激素受体的蛋白表达水平.结果:人急性淋巴细胞白血病细胞系MOLT-4、CCRF-CEM和CEM-C1中HOTAIR的表达显著高于人正常骨髓基质HS-5细胞(P<0.01).在CEM-C1细胞中干扰HOTAIR表达后,细胞活力降低,细胞增殖被抑制并发生凋亡,地塞米松抑制CEM-C1增殖的作用被增强,糖皮质激素受体表达上调(P<0.01).结论:长链非编码RNA HOTAIR增强急性淋巴细胞白血病的活力,促进其增殖并抑制其凋亡,该作用可能与其抑制糖皮质激素受体的表达有关.

目的:优化瑶药过墙风总黄酮提取工艺,为其开发和利用提供参考.方法:在单因素试验筛选基础上,以乙醇浓度、提取温度和提取时间为自变量,总黄酮得率为因变量,然后根据星点设计原理进行试验,对结果进行多元线性回归和二项式方程拟合,采用效应面法优化过墙风总黄酮的提取工艺.结果:过墙风总黄酮最优提取工艺为乙醇浓度54.76％,提取温度83.92℃,提取时间102.64 min,固液比为1:20,提取2次.结论:本实验星点设计-效应面法优化过墙风总黄酮提取工艺简便可行,预测结果可靠,适于工业化生产.

To explore the protective effects and its mechanism of Clerodendrum philippinum Schauer var.simplex Mlodenke total flavonoids (CPTF) on acute liver injury induced by carbon tetrachloride (CCl4) in mice.60 mice were randomly divided into normal group,model group,silymarin group (150 mg/kg),CPTF-high,medium and low dose group (200,100,50 mg/kg).The normal group and model group were given equivoluminal distilled water,and other groups were given the corresponding dose of drugs (25 mL/kg) for 10 days.After 2 h of the last administration,the mice of all groups except the normal group were injected intraperitoneally with 0.1％ CCl4 peanut oil solution (10 mL/kg).16 h later,the blood was taken by removing eye balls,and the liver tissue was collected after killing the mice.The serum contents of alanine aminotransferase (ALT),aspartate aminotransferase (AST),total superoxide dismutase (T-SOD),malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) were measured by biochemical method;the levels of tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β) and interleukin-6 (IL-6) in liver tissue were detected by enzymelinked immunosorbent assay (ELISA);janus kinase 2 (JAK2),phosphorylated JAK2 (p-JAK2),signal transducer and activator of transcription 3 (STAT3) and phosphorylated STAT3 (p-STAT3) in liver tissue were detected by Western blot;the histopathological changes of liver were observed by HE staining.The results showed that CPTF could significantly improve the liver tissue lesion,decrease the levels of AST,ALT and MDA in serum (P ＜ 0.05),enhance the activity of T-SOD and GSH-Px (P ＜ 0.05),inhibit the hepatic levels of TNF-α,IL-1β and IL-6 (P ＜ 0.05),and inhibit the expression of p-JAK2 and p-STAT3 (P ＜ 0.05).In summary,CPTF had a protective effect on CCl4-induced acute liver injury in mice,which could reduce the degree of liver injury,and its mechanism may be related to anti-oxidation,inhibition of inflammation and regulation of JAK2/STAT3 signaling pathway.

目的:研究七味净肝灵对二甲基亚硝胺(DMN)诱导肝纤维化大鼠的保护作用及其作用机制.方法:90只大鼠随机分为正常对照组、模型组、秋水仙碱(0.12 mg/kg)阳性对照组和七味净肝灵高(8 000 mg/kg)、中(4000mg/kg)、低(2 000 mg/kg)剂量组,每组15只.除正常对照组腹腔注射相应体积的生理盐水外,其余组均按1.5mL/kg腹腔注射0.5％ DMN溶液,1次/2 d,同时每天灌胃相应药物进行干预,正常对照组与模型组灌胃相应体积生理盐水,共40 d.实验期间观察并记录大鼠的活动、体质量、外观及毛发变化情况.末次给药后,禁食不禁水16h,随后眼球取血,收集血清及肝脏.采用生化法检测各组大鼠血清中AST、ALT、ALB、TBIL水平;ELISA检测各组大鼠肝组织中PCⅢ、LN、Ⅳ-C、HA含量;Western blot检测各组大鼠肝组织中α-SMA、TGF-β1蛋白表达水平;HE染色进行肝组织病理学检查.结果:与模型组比较,七味净肝灵高、中剂量组大鼠血清AST、ALT、TBIL水平显著降低,ALB水平显著升高(P＜0.05或P＜0.01);肝组织中PCⅢ、LN、Ⅳ-C、HA含量显著降低(P＜0.05或P＜0.01),α-SMA、TGF-β1蛋白表达水平显著降低(P＜0.05或P＜0.01);病理切片显示,七味净肝灵高、中剂量组大鼠肝纤维化病变程度较模型组明显减轻.结论:七味净肝灵对DMN诱导的大鼠肝纤维化具有显著的改善作用,其作用机制可能与抑制TGF-β1有关.

Objective: To study the protective effect and mechanism of total flavonoids of Polygonum Chinense L. (TFP) on the acute hepatic injury in mice induced by D-galactosamine (D-Gal N). Methods: 60 mice were randomly divided into the control group, the model group, silymarin group (200 mg/kg) and TFP groups (400 mg/kg, 200 mg/kg, 100 mg/kg), mice were orally administered once a day for 7 consecutive days. One hour after the last administration, except the control group, mice in other groups were given intraperitoneal injection of D-Gal N (500 mg/kg) to establish acute liver injury model in mice. Then mouse serum and liver tissues were collected on 16 th hour after models were established. Contents of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TBIL), total superoxide dismutase (T-SOD), malondialdehyde (MDA) and glutathione Oxidase (GSH-Px) of mice serum were detected. Expression levels of Th1 cytokines TNF-α, IFN-γ and Th2 cytokines IL-10, IL-4 were detected by ELISA. Pathological changes of mice liver tissues were observed by HE staining. Results: Compared with the control group, levels of ALT, AST, TBIL, MDA, TNF-α and IFN-γ significantly increased, whereas activities of GSH-Px, T-SOD, IL-10 and IL-4 decreased (P ＜0. 01). After interventions of TFP (400 mg/kg, 200 mg/kg) on the acute liver injury, expression levels of ALT, AST, TBIL, MDA, TNF-α and IFN-γ in serum or liver tissues were decreased (P ＜0. 05, P＜0. 01), activities of GSH-Px, T-SOD, IL-10 and IL-4 were increased (P ＜0. 05, P ＜0. 01). And pathological changes showed that TFP (400 mg/kg, 200 mg/kg significantly improved the pathological characteristics of liver tissues. Conclusion: TFP plays the significant liver protection role, and its mechanism may be related to anti-oxidation and regulation of liver tissue Th1/Th2 balance.

细胞自噬是真核生物内对蛋白质或受损细胞器等细胞组分进行周转的过程,其对维持细胞稳态具有重要作用.自发现以来,研究者对这一重要的细胞过程及其基础机制展开了大量研究并取得了重大进展.其中,细胞自噬与癌症的发生发展有着密切联系.本文主要就自噬对癌症发生的影响展开概述,为全面理解自噬及其与癌症关系提供新思路.

Aim Based on the effect of total flavonoids extracted from Polygonum perfoliatum L.(TFP) against dimethylnitrosamine(DMN)-induced hepatic fibrosis(HF), to investigate the anti-fibrotic mechanism of TFP.Methods Ninety SD rats were divided into normal group, model group, colchicines(0.1 mg·kg-1) group, and TFP(200, 100, 50 mg·kg-1) group.Except the rats of normal group, other rats were injected intraperitoneally with volume fraction 0.5% DMN solution(2 mL·kg-1) for eight weeks, once every two days.From the first day of modeling, each administration group was given the corresponding dose of drugs to intervene, and the normal group and model group were given an equal volume of solvent, once a day.At the end of the eighth week, the blood and liver tissues were collected.Liver tissue was taken at a fixed position, and the degree of liver tissue was observed by HE staining.The contents of serum ALT, AST, SOD and MDA were measured using colorimetric method;the levels of serum HA, LN, PCⅢand Ⅳ-C were detected using enzyme-linked immunosorbent assay(ELISA);the levels of TNF-α, IL-1β and IL-6 in liver tissues were detected by ELISA;the expression of α-SMA, TGF-β1, p-JAK2 and p-STAT3 were detected by Western blot.Results Compared with the model group, TFP(200, 100, 50 mg·kg-1) could improve the liver tissue lesions, reduce the expression of ALT, AST, HA, LN, PCⅢ, IV-C and MDA, increase SOD activity, reduce the levels of TNF-α, IL-1β and IL-6, and inhibit the expression of α-SMA, TGF-β1, JAK2, STAT3, p-JAK2 and p-STAT3.Conclusion TFP could inhibit DMN-induced HF of rats, which may be involved with antioxidant and inhibiting expression of TGF-β1, JAK2/STAT3 signaling pathway and inflammatory response.

OBJECTIVE To study the protective effects of Polygonum perfoliatum on hepatic fibrosis induced by DMN in rats based on TGF-β1/Notch signaling pathway.METHODS Rats were injected intraperitoneally with 0.5％ DMN solution at 1.6 mL·kg-1,three times a week for 42 days to establish rat liver fibrosis models.At the same time,colchicines (0.1 g· kg-1) and Polygonum perfoliatum (10,5,2.5 g · kg-1) were administered for intervention once daily.After the last administration,rats were fast for 16 h,and then the serum and liver tissues were collected.ALT and AST in serum were measured by biochemical method.ELISA was used to measure the contents of PC Ⅲ,Ⅳ-C,LN and HA in liver tissues.Western-blot was used to detect the expressions of α-SMA,TGF-β1,Notch-1 and E-cadherin in liver tissues.Histopathological changes of liver were observed by HE staining.RFSULTS Compared with the model group,the levels of ALT and AST in the rat serum of Polygonum perfoliatum groups were significantly decreased (P＜0.05 or P＜0.01).The levels of PC Ⅲ,Ⅳ-C,LN and HA in liver tissues of Polygonum perfoliatum groups (10,5 g· kg-1) were decreased (P＜0.05 or P＜0.01).The expressions of TGF-β1,Notch-1 and α-SMA protein in liver tissues were decreased,while the expression of E-cadherin protein was increased (P＜0.05 or P＜0.01).The results of HE staining showed that the hepatic fibrosis of the rats of Polygonum perfoliatum groups was improved to different degrees.CONCLUSION Polygonum perfoliatum has significant protective effects on hepatic fibrosis rats,and its mechanism may be related with the regulation of TGF-β1/Notch signaling pathway.

The polysaccharides of Dicliptera chinensis (L.) Juss. (DCP-1 and DCP-2) were extracted and isolated using the methods of water extract–ethanol precipitate and sephadex column chromatography and characterized by gel permeation chromatography (GPC), Fourier transform infrared spectrometry (FT-IR) and gas chromatography (GC), respectively. The antioxidant activity of DCPs was evaluated by scavenging activity of DPPH, hydroxyl, superoxide anion and ABTS radical. Moreover, the anti-aging activity of DCP-2 was investigated using an aging model-induced by D-galactose (D-gal) in mice. The results show that the weight average molecular weight (Mw) of DCP-2 was 2 273Da with a narrow polydispersity index of 1.01, and it was a heteropolysaccharide and consisted of glucose, galactose, arabinose, rhamnose and mannose with a molar ratio of 3.20:2.54:1.69:1.58:1.00. DCP-2 had stronger antioxidant activity against DPPH, hydroxyl, superoxide anion and ABTS radical, while DCP-1 had hardly any antioxidant activity and DCP had weaker antioxidant activity. Furthermore, DCP-2 can enhance antioxidant capacity and had anti-aging activity against D-gal induced aging mice. These results proposed that DCP-2 might be developed as a potential functional food with the activity of anti-aging.

Objective:To study the liver-protecting function and mechanism of Qiwei Jingganling based on JAK2/STAT3 signal pathway.Methods:60 mice were randomly divided into normal control group,model group,Silybin group(0.5 g/kg)and Qiwei Jingganling high-,medium-and low-dose(20,10,5 g/kg)groups,with ten rats in each groups.The normal control group and the model group were injected with water daily,and the other groups were given corresponding drugs with intragastric administration once a day for 7 d.After last administration,the normal control group was injected with peanut oil,the other groups were injected with 0.12％ CC14 mix peanut oil (10 mL/kg)to induce acute liver injury model.Lasting for 24 h,the content of ALT,AST,MDA,SOD,GSH-Px in serum and the expression of TNF-α,IL-1β,IL-6 in liver tissue were measured,and the protein expression of SOCS3,p-JAK2 and p-STAT3 in liver tissue were detected by Western blot;the liver tissue pathological change was deteced by HE staining method by using microscope observation.Results:Compared with model group,the activities of MDA,AST and ALT and in serum of Qiwei Jingganling groups were decreased significantly,the activities of SOD and GSH-PX in serum of Qiwei Jingganling groups were increased significantly;the protein expression of TNF-α,IL-1β,IL-6,p-JAK2 and p-STAT3 in liver tissue were decreased,the expression of SOCS3 in liver tissue was increased significantly;and the pathological sections showed that the hepatic injury were relieved clearly.Conclusion:Qiwei Jingganling has a significant hepatoprotective effect,of which mechanism may be related to anti-oxidation and inhibition of inflammatory response,and regulate the JAK2/STAT3 signaling pathway by mediating SOCS-3.

目的:以二甲基亚硝胺(DMN)诱导的大鼠肝纤维化为模型,研究狗肝菜多糖对DMN诱导的肝纤维化大鼠TGF-β1/Smads信号通路的影响.方法:将96只大鼠随机分为对照组、模型组、狗肝菜多糖(300 mg/kg,200 mg/kg,100 mg/kg)组和秋水仙碱组(0.12mg/kg).除对照组以外,其余各组大鼠均腹腔注射0.5％ DMN溶液(1.5 ml/kg),隔天1次,在首次注射DMN同时,各组大鼠灌胃给予相应的药物,而正常组和模型组灌胃给予相应体积的生理盐水溶液,每天给药1次,连续5周,5周末眼球取血,脱臼致死,收取肝脏.生化法检测血清中碱性磷酸酶(ALP)、谷氨酰氨基转移酶(GGT)水平;酶联免疫法检测(ELISA)血清中透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PCⅢ)和Ⅳ型胶原(Ⅳ-C)含量.苏木精-伊红染色(HE染色)进行肝组织病理学检查;蛋白质印迹法(Westem blot)检测肝组织a-平滑肌肌动蛋白(a-SMA)、转化生长因子β1(TGF-β1)和Smad2/3/7的蛋白表达情况.结果:与模型组比较,经狗肝菜多糖(300 mg/kg,200 mg/kg,100 mg/kg)组干预后,血清中ALP、GGT、HA、LN、PCⅢ和Ⅳ-C水平显著降低;肝组织中a-SMA、TGF-β1、Smad2/3的蛋白表达被抑制,而Smad7的表达得到促进;病理切片显示肝纤维化大鼠肝组织炎症坏死程度和肝纤维化病变程度较模型组明显减轻.结论:狗肝菜多糖具有抗DMN诱导的大鼠肝纤维化的作用,其机制与TGF-β1/Smads信号通路相关因子的表达有关.

目的:研究金花茶多糖对四氯化碳(CCl4)诱发的小鼠急性肝损伤的保护作用及其作用机制.方法:60只小鼠随机分为正常对照组、模型组、联苯双酯组(120 mg/kg)和金花茶多糖组(200 mg/kg,100mg/kg,50 mg/kg).除正常对照组及模型组外,各治疗组按20 ml/kg剂量灌胃给药,每天按时灌胃1次,共7天.末次给药后,除正常对照组外,其余组腹腔注射0.12％ CCl4花生油(10 ml/kg)建立急性肝损伤模型.禁食不禁水24h后,眼球取血,收集肝脏.生化法测定血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的活性或含量;酶联免疫吸附法(ELISA)检测肝组织中肿瘤坏死因子-α (TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)表达水平和一氧化氮(NO)含量;蛋白质印迹法(Western blot)检测肝组织NF-κB蛋白表达情况;HE染色肝切片,显微镜观察肝组织病理改变.结果:与CCl4模型组相比,金花茶多糖剂量组(200mg/kg,100 mg/kg)小鼠血清中ALT、AST及MDA含量降低,SOD及GSH-Px的活性升高;肝组织中NO含量显著降低;肝组织中TNF-α、IL-1β、IL-6及NF-κB表达水平呈下降趋势;病理切片表明金花茶多糖剂量组(200 mg/kg,100 mg/kg)小鼠肝损伤显著减轻.而金花茶多糖50 mg/kg剂量组对CCl4诱发的小鼠急性肝损伤无明显改善作用.结论:金花茶多糖对CCl4诱导的小鼠急性肝损伤有明显的保护作用,其机制可能与抗氧化应激,清除自由基代谢产物,抑制脂质过氧化反应及抗炎有关.

Aim To study the effects of Dicliptera chinensis polysaccharide(DCP)on alcoholic fatty liver disease(AFLD)in rats based on anti-inflammation and antioxidation.Methods 60 rats were randomly divid-ed into six groups:control group,model group,silybin group and DCP of high,medium and low dose groups. The control group was fed with normal diet, other groups were fed with high sugar and high fat diet,and given 5% alcohol 5 mL·kg-1 by gavage.The alcohol consistency increased 5%every week until AFLD mod-els in rats were made after 7 weeks.Except control group,other groups were fed with high sugar and high fat diet,and given 35% alcohol 5 mL · kg-1 and DCP.All rats were killed after five weeks,and blood and liver tissues were collected.The activity of alanine aminotransaminase (ALT),aspartate aminotransferase (AST ), alkaline phosphatase (AKP ), triglyceride (TG),total cholesterol (TC ),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cho-lesterol(HDL-C)in serum were detected by using bio-chemical method. The contents of malondialdehyde (MDA),superoxide dismutase (SOD),reduced gluta-thione(GSH)in liver tissues were detected.The con-tents of tumor necrosis factor-α(TNF-α),interleukin-6 (IL-6 )and transforming growth factor-β1 (TGF-β1 ) were determined by enzyme-linked immunosorbent as-say(ELISA)in liver tissues.The liver tissues were ob-tained and histologic analysis was done through HE. Results DCP reduced the activity or content of ALT, AST,AKP,TG,TC,LDL-C,HDL-C,TNF-α,IL-6, TGF-β1 in serum and liver tissues of rats(P<0.05 ), and increased the activity or content of HDL-C,SOD and GSH (P<0.05 ).DCP could remarkably inhibit the NF-κB expression in liver tissues(P<0.01 ).The pathological examination indicated that DCP could ob-viously alleviate the inflammation and fat denaturation of the liver cells.Conclusion DCP can inhibit the de-velopment of AFLD.The mechanism may be related to antioxidation,free radical scavenging, inhibition of lipidperoxidation,anti-inflammation,and inhibition of the TGF-β1 and NF-κB expression.