Zuogui Jiangtang Jieyu Formula ameliorating hippocampal neuronal apoptosis in diabetic rats with depression by inhibiting JNK signaling pathway

Objective To investigate the effect of Zuogui Jiangtang Jieyu Formula(左归降糖解郁方,ZJJF)on hippocampal neuron apoptosis in diabetic rats with depression and to ascertain whether its mechanism involves the regulation of JNK signaling pathway.Methods(i)A total of 72 specific pathogen-free(SPF)grade male Sprague Dawley(SD)rats were randomly divided into six groups,with 12 rats in each group:control,model,metformin(Met,0.18 g/kg)+fluoxetine(Flu,1.8 mg/kg),and the high-,medium-,and low-ZJJF dosages(ZJJF-H,20.52 g/kg;ZJJF-M,10.26 g/kg;ZJJF-L,5.13 g/kg)groups.All groups except control group were injected once via the tail vein with streptozotocin(STZ,38 mg/kg)combined with 28 d of chronic unpredictable mild stress(CUMS)to establish diabetic rat models with depression.During the CUMS modeling period,treatments were administered via gavage,with control and model groups receiving an equivalent volume of distilled water for 28 d.The efficacy of ZJJF in reducing blood sugar and alleviating depression was evaluated by measuring fasting blood glucose,insulin,and glycated hemoglobin levels,along with behavioral assessments,including the open field test(OFT),forced swim test(FST),and sucrose preference test(SPT).Hippocampal tissue damage and neuronal apoptosis were evaluated using hematoxylin-eosin(HE)staining and terminal deoxynucleotidyl transferase-mediated dUTP nickend labeling(TUNEL)staining.Apoptosis-related proteins Bax,Bcl-2,caspase-3,and the expression levels of JNK/Elk-1/c-fos signaling pathway were detected using Western blot and real-time quantitative polymerase chain reaction(RT-qPCR).(ii)To further elucidate the role of JNK signaling pathway in hippocampal neuronal apoptosis and the pharmacological effects of ZJJF,an additional 50 SPF grade male SD rats were randomly divided into five groups,with 10 rats in each group:control,model,SP600125(SP6,a JNK antagonist,10 mg/kg),ZJJF(20.52 g/kg),and ZJJF(20.52 g/kg)+Anisomycin(Aniso,a JNK agonist,15 mg/kg)groups.Except for control group,all groups were established as diabetic rat models with depression,and treatments were administered via gavage for ZJJF and intraperitoneal injection for SP6 and Aniso for 28 d during the CUMS modeling period.Behavioral changes in rats were evaluated through the OFT,FST,and SPT,and hippocampal neuron damage and apoptosis were observed using HE staining,Nissl staining,TUNEL staining,and transmission electron microscopy(TEM).Changes in apoptosis-related proteins and JNK signaling pathway in the hippocampal tissues of rats were also analyzed.

Timosaponin AⅢ induces drug-metabolizing enzymes by activating constitutive androstane receptor (CAR) via dephosphorylation of the EGFR signaling pathway

The current study aimed to assess the effect of timosaponin AⅢ(T-AⅢ)on drug-metabolizing enzymes during anticancer therapy.The in vivo experiments were conducted on nude and ICR mice.Following a 24-day administration of T-AⅢ,the nude mice exhibited an induction of CYP2B10,MDR1,and CYP3A11 expression in the liver tissues.In the ICR mice,the expression levels of CYP2B10 and MDR1 increased after a three-day T-AⅢ administration.The in vitro assessments with HepG2 cells revealed that T-AⅢ induced the expression of CYP2B6,MDR1,and CYP3A4,along with constitutive androstane receptor(CAR)activation.Treatment with CAR siRNA reversed the T-AⅢ-induced increases in CYP2B6 and CYP3A4 expression.Furthermore,other CAR target genes also showed a significant increase in the expression.The up-regulation of murine CAR was observed in the liver tissues of both nude and ICR mice.Subsequent findings demonstrated that T-AⅢ activated CAR by inhibiting ERK1/2 phosphorylation,with this effect being partially reversed by the ERK activator t-BHQ.Inhibition of the ERK1/2 signaling pathway was also observed in vivo.Additionally,T-AⅢ inhibited the phosphorylation of EGFR at Tyr1173 and Tyr845,and suppressed EGF-induced phosphorylation of EGFR,ERK,and CAR.In the nude mice,T-AⅢ also inhibited EGFR phosphorylation.These results collectively indicate that T-AⅢ is a novel CAR activator through inhibition of the EGFR pathway.

MEK/ERK signaling pathway in apoptosis of SW620 cell line and inhibition effect of resveratrol

Objective:To study the involvement of MAPK MEK/ERK signaling transduction pathway in the apoptosis process of SW620 tumor cell line and the inhibition effect of resveratrol.Methods:SW620 cell lines were divided into 5 groups,namely,control group.PD98059 group,low-dose resveratrol group,mid-dose resveratrol group and high-dose resveratrol group.The inhibition rate of cell proliferation was detected by MTT method.The expression of apoptotic molecules and MEK/ERK signaling pathway related proteins were assayed by realtime PCR and Western blotting.Results:Compared with control group,the proliferation of cells treated with resveratrol was significantly inhibited.In the case of apoptotic molecules,the expression of Bax,Caspase 3 and Caspase 9 was increased significantly while the expression of anti-apoptotic molecule Bcl2 was decreased significantly in resveratrol groups with a dosedependent manner.In the case of molecules in MEK/ERK signaling pathway,the expression of Ras,Raf,MEK and ERKl/2 was decreased significantly in resveratrol groups with a dose-dependent manner.Conclusions:PD98059 and resveratrol can effectively inhibit the proliferation of SW620 through inhibiting the MEK/ERK signaling pathway.

Endogenous hydrogen sulfide and ERK1/2-STAT3 signaling pathway may participate in the association between homocysteine and hypertension

Background Homocysteine(Hcy)is a risk factor for hypertension,although the mechanisms are poorly understood.Methods We first explored the relationship between Hcy levels and blood pressure(BP)by analyzing the clinical data of primary hypertensive patients admitted to our hospital.Secondly,we explored a rat model to study the effect of Hcy on blood pressure and the role of H2S.An hyperhomocysteinemia(HHcy)rat model was induced to explore the effect of Hcy on blood pressure and the possible mechanism.We carried out tissue histology,extraction and examination of RNA and protein.Finally,we conducted cell experiments to determine a likely mechanism through renin-angiotensin-aldosterone system(RAAS)and extracellular signal-regulated kinase 1/2(ERK1/2)signaling pathway.Results In primary hypertensive inpatients with HHcy,blood pressure was significantly higher as compared with inpatient counterparts lacking HHcy.In the rat model,blood pressure of the Wistar rats was significantly increased with increases in serum Hcy levels and decreased after folate treatment.Angiotensin converting enzyme 1(ACE1)expression in the Wistar Hcy group was enhanced comparing to controls,but was decreased in the Wistar folate group.Angiotensin II receptor type 1(AGTR1)levels in the kidney tissue increased in the Wistar folate group.Both serum H2S and kidney cystathionineγ-lyase decreased with elevated levels of serum Hcy.In vitro,increased concentrations and treatment times for Hcy were associated with increased expression of collagen type 1 and AGTR1.This dose and time dependent response was also observed for p-STAT3 and p-ERK1/2 expression.Conclusion Endogenous H2S might mediate the process of altered blood pressure in response to changes in serum Hcy levels,in a process that is partly dependent on activated RAAS and ERK1/2-STAT3 signaling pathway.

Study on the role of Cathepsin B and JNK signaling pathway in the development of cerebral aneurysm

Objective: To investigate the correlation between JNK signal and the apoptosis of VSMC as well as the expression of Cathepsin B and to explore the role of JNK signal in the development of cerebral aneurysm. Methods: Rat models of cerebral aneurysm were established and histopathologic changes of cerebral aneurysm and the apoptosis of VSMC were analyzed. Rat models were respectively subject to subcutaneous injection of Cathepsin B si RNA and JNK inhibitor SP600125. Western blot technique was used to detect the expression of proteins like Cathepsin B, Caspase-3, and p-JNK. Spearman's rho was used to examine the correlation between p-JNK and Cathepsin B, as well as the expression of relevant proteins. Results:The success rate of modeling rats with cerebral aneurysm was 88.75%. After the respective injection of Cathepsin B si RNA, SP600125 and their combination, the cell densities of VSMC of rats with cerebral aneurysm all increased significantly(P<0.05 or P<0.01), but the apoptosis rate of VSMC decreased significantly(P<0.01). Compared with normal rats, the expression of Cathepsin B, Caspase-3 and p-JNK in cerebral aneurysm models increased significantly. Effectively intervening Cathepsin B genes with Cathepsin B si RNA could significantly inhibit the expression of Cathepsin B and Caspase-3, but hardly influence the expression of p-JNK. JNK inhibitor SP600125 had no influence on the expression of Cathepsin B and Caspase-3, but effectively inhibited the expression of p-JNK. In cerebral aneurysm tissues, positive correlation was observed between the expression of p-JNK and Cathepsin B, the correlation coefficient was r=0.640. Conclusion: After the attack of cerebral aneurysm, proteins like Cathepsin B, Caspase-3 and p-JNK are all involved in the apoptosis of VSMCs. This process may be realized by Cathepsin B which activates the apoptosis mechanism of Caspase-3 and mediate the apoptosis of VSMC through the JNK signaling pathway. Therefore, silencing Cathepsin B gene or inhibiting the conduction through JNK signaling pathway can mitigate the apoptosis of vascular smooth muscle cells in cerebral aneurysm.

Acupuncture at Back-Shu point improves insomnia by reducing inflammation and inhibiting the ERK/NF-κB signaling pathway

BACKGROUND Insomnia is a disease where individuals cannot maintain a steady and stable sleep state or fail to fall asleep.Western medicine mainly uses sedatives and hypnotic drugs to treat insomnia,and long-term use is prone to drug resistance and other adverse reactions.Acupuncture has a good curative effect and unique advantages in the treatment of insomnia.AIM To explore the molecular mechanism of acupuncture at Back-Shu point for the treatment of insomnia.METHODS We first prepared a rat model of insomnia,and then carried out acupuncture for 7 consecutive days.After treatment,the sleep time and general behavior of the rats were determined.The Morris water maze test was used to assess the learning ability and spatial memory ability of the rats.The expression levels of inflammatory cytokines in serum and the hippocampus were detected by ELISA.qRTPCR was used to detect the mRNA expression changes in the ERK/NF-κB signaling pathway.Western blot and immunohistochemistry were carried out to evaluate the protein expression levels of RAF-1,MEK-2,ERK1/2 and NF-κB.RESULTS Acupuncture can prolong sleep duration,and improve mental state,activity,diet volume,learning ability and spatial memory.In addition,acupuncture increased the release of 1L-1β,1L-6 and TNF-αin serum and the hippocampus and inhibited the mRNA and protein expression of the ERK/NF-κB signaling pathway.CONCLUSION These findings suggest that acupuncture at Back-Shu point can inhibit the ERK/NF-κB signaling pathway and treat insomnia by increasing the release of inflammatory cytokines in the hippocampus.

The role of ERK1/2 signaling pathway in coronary microembolization-induced rat myocardial inflammation and injury

Objectives In this work,we explore the effect of atorvastatin on myocardial apoptosis and caspase-8 acti- vation after coronary microembolization(CME) in rats. Methods Fifty rats were randomly divided into five groups; the coronary microembolization(CME) group,the sham-operated (sham) control group,the gastric lavage control group, the atorvastatin lavage group,and the caspasse-8 inhibitor (N-acetyl-Ile-Glu-Thr-Asp-CHO,abbreviated as CHO) group,with 10 rats for each group.A microembolization ball was injected through the left ventricle for constructing the CME model.Animals in the sham control group were given an injection of physiological saline instead of the microembolization ball.Seven days before the operation,the atorvastatin group underwent gastric lavage with 20 mg/kg of atorvastatin once a day.Gastric lavage control animals underwent gastric lavage with an equivalent dose of physiological saline instead of the atorvastatin.Animals in the CHO group were given an intraperitoneal injection of 10 mg/kg of CHO 30 min before the operation.Six hours after the operation,cardiac ultrasonic detection was conducted on each group to measure the cardiac function indexes.TUNEL(Terminal-deoxynucleoitidyl transferase mediated dUTP nick end labeling) assays were used to measure myocardial apoptosis,and western blots were used to quantify the expression levels of activated caspase-3 and -8.Results(1) The echocardiographic parameters showed that,compared to the sham control animals,the left ventricular ejection fraction(LVEF) of the CME group was significantly decreased(P【0.05).In addition, cardiac sonography revealed a decrease in the left ventricular shortening fraction(FS) and cardiac output(CO), but an increase in the left ventricular end-diastolic dimension (LVEDd).Compared to the CME group,the atorvastatin and CHO groups exhibited significantly improved cardiac function (P【0.05).(2) When compared with the sham control,the myocardical apoptotic rate of the CME group,as well as the levels of activated caspase-3 and-8,increased significantly (P【0.05).The myocardial apoptotic rate,as well as the levels of activated caspase-3 and caspase-8 in the atorvastatin and CHO groups,decreased significandy(P【0.05) in comparison to the CME group.Conclusions The atorvastatin pretreatment clearly suppressed post-CME myocardial apoptosis and improved cardiac function.The most likely mechanism for these effects is the blockade of the myocardial death receptor -mediated apoptosis pathway.

Irisin Attenuates Osteoarthritis by Inhibiting Apoptosis of Osteocytes Through Activating Erk Signaling Pathway

Osteoarthritis(OA)is an inflammatory disease involving the joints that is prevalent in the global aging population.The purpose of this study is to determine whether irisin can attenuate osteoarthritis(OA)progression in anterior cruciate ligament transection(ACLT)mice models and the mechanism of irisin therapy effect on OA by increase the resistance of apoptosis in MLO-Y4 cells induced by mechanical stretch in vitro.Methods For in vivo study,3-month-old male C57BL/6 J mice were randomized to three groups,sham-operated,anterior cruciate ligament transection(ACLT)-operated treated with vehicle,and ACLT-operated treated with irisin by intraperitoneal injection once a week.Cartilage erosion was observed by HE staining.Osteoarthritis Research Society International(OARSI)scores were evaluated according to the safranin O stai-ning.The microstructure of tibia cortical bone,trabecular bone,and subchondral bone was analyzed by micro-CT and the bone histomorphometry has been administrated including mineral apposition rate(MAR).Edu staining and cck-8 were used for the detection of the proliferation of MLO-Y4 cells.For mechanical stress,cells were seeded on the collagen-I coated chamber subjected with a peak biaxial stretch of 20%at 1 Hz for 16 hours to induce apoptosis.Flow cytometry was used for the detection of apoptosis and cell cycle.TUNNEL was used for staining the apoptotic cells and rt-PCR was applied for quantifying the expression of mRNA such as Bax,Bcl-2,SOST,c-myc,Opg.Western blot was utilized to confirm the mechanism of how irisin decrease the osteocyte apoptosis.Results In vivo,irisin can attenuate articular cartilage degeneration.Irisin maintains the proportion of hyaline cartilage and calcified cartilage and keep fewer cartilage erosions in ACLT-operated mice.For immunohistochemical(IHC)staining,irisin reduced the expression of caspase3,Bax and matrix metalloproteinase-13 in both cartilage and subchondral bone.Irisin-treated ACLT group shows higher Trabecular number(Tb.N)and bone volume fraction(BV/TV)compared to the vehicle-treated ACLT group.In vitro, irisin significantly increased the proliferation of MLO-Y4 cells detected by Edu and Ki67 staining,and irisin can protect the cells from both mechanical stretchinduced apoptosis detected by FITC-PI flow cytometry and maintain the cell activity by regulating the expression of Bax,Bcl-2,and c-myc.Transcriptome sequencing shows that irisin significantly activates the MAPK signaling pathway and we confirm the result by western blot:irisin effectively activates the Erk signaling pathway through phosphorylation and has a certain activation effect on p38 signaling pathway,no activation was observed for FAK signaling pathway.Conclusions Irisin can attenuate the progression of OA by decrease the apoptosis of osteocyte,which can improve the microarchitecture of subchondral bone.Erk pathway activation plays an important role in reducing the apoptosis of osteocyte.

To investigate the effects of butylphthalide on reducing neuronal apoptosis in rats with cerebral infarction by inhibiting the JNK/P38 MAPK signaling pathway

Objective:To investigate the effects of butylphthalide on reducing neuronal apoptosis in rats with cerebral infarction by inhibiting the JNK/P38 MAPK signaling pathway.Methods:Forty-eight SD male rats were divided into DZ group(control group),CI group(model group)and NBP group(butylphthalide group).Rats in CI group and NBP group were used to establish cerebral infarction models.NBP group used NBP.The solution(80 mg/(kg?d))was administered orally,and the remaining two groups were administered with the same volume of peanut oil.After 14 consecutive days of treatment,the Zea Longa score was used to evaluate the neurological function of DZ,CI and NBP rats.Scoring,TTC staining was used to observe the cerebral infarction volume of rats in DZ group,CI group and NBP group,HE staining was used to observe the pathological morphology of brain tissue in DZ group,CI group and NBP group.Neuronal apoptosis,Western blot was used to detect the expression of p-JNK and p-p38MAPK in brain tissues of DZ group,CI group and NBP group.Results:The neurological function of the rats in the CI group was higher than that in the DZ group,and the difference was statistically significant(P<0.05).The neurological function score of the rats in the NBP group was reduced compared with the CI group,and the difference was statistically significant(P<0.05).The cerebral infarction volume in the group was 35.56%higher than that in the DZ group,and the difference was statistically significant(P<0.05).The minor infarct volume in the NBP group was 21.59%,which was less than that in the CI group,and the difference was statistically significant(P<0.05).Nerve cells are neatly sorted,with a large number.The gap between blood vessels and interstitial tissue in the CI group is enlarged,the cells are severely contracted,and the neuron structure is incomplete.Compared with the CI group,the NBP group has reduced neuron contraction and increased number;The dead nerve cells were brown.The apoptosis rate of nerve cells in the CI group was 79.65%higher than that in the DZ group was 5.82%.The difference was statistically significant(P<0.05).The nerve cell apoptosis rate in the NBP group was 30.23%.Compared with CI group,the difference was statistically significant(P<0.05);Western blot results showed that p-JNK and p-p38MAPK protein expression in CI group was higher than that in DZ group,and the difference was statistically significant(P<0.05).The levels of p-JNK and p-p38MAPK proteins in the NBP group were lower than those in the CI group.There was statistically significant(P<0.05).Conclusion:Butylphthalide can improve neurological damage,reduce apoptotic nerve cells,and reduce infarct volume in rats with cerebral infarction,which is related to the inhibition of JNK/P38 MAPK pathway expression.

Maleylated-BSA induces TNF-α production through the ERK and NF-κB signaling pathways in murine RAW264.7 macrophages

Ligands for macrophage scavenger receptors are reported to induce a wide range of host cell responses, including the production of inflammatory cytokines;however, the underlying mechanisms have not yet been fully understood and which remain obscure. In this study, we have examined the effect of maleylated bovine serum albumin (maleylated-BSA), a well-known ligand of the scavenger receptor, on the murine macrophage cell line RAW264.7. Maleylated-BSA strongly induced the production of tumor necrosis factor-α (TNF-α) and induced phosphorylation of extracellular signal-regulated kinase (ERK) and NF-kB p65. We also observed that maleylated-BSA-induced TNF-α production was blocked by the ERK inhibitor U0126. Together, these data demonstrates that maleylated-BSA- induced production of TNF-α requires the ERK/NF-κB signaling cascade in murine RAW- 264.7 macrophages.

Role of JNK signaling pathway in sensitivity to radiotherapy of nasopharyngeal carcinoma

Objective:The aim of the study was to investigate the effect of c-Jun N-terminal protein kinase(JNK) signaling pathway on influencing the sensitivity to radiotherapy of human nasopharyngeal carcinoma CNE cells.Methods:Human nasopharyngeal carcinoma CNE multicellular spheroids(MCS) were constructed with three dimensional cell culture methods.Western blot was employed to analyze the activity of JNK signaling pathway in MCS after X-ray irradiation,and the expression of caspase-3 protein before and after using SP600125(a special inhibitor of JNK).X-ray induced cell apoptosis in MCS before and after treated with SP600125 were detected by TUNEL.Results:The level of JNK phosphorylation in MCS was a dynamic course after radiation,and there was a phosphorylation peaks at 2 h later,the apoptotic rate of MCS(P < 0.05) and the expression of caspase-3 protein(P < 0.05) were significantly increased after treated with SP600125.Conclusion:The transient activation of JNK played a important role in sensitivity to radiotherapy of CNE MCS via mediating survival signals,blocking this pathway accelerate cell apoptosis,which may be related to the increased expression of caspase-3.

ERK和JNK信号转导通路与疾病关系的研究进展

ERK信号通路激活能够促进细胞增殖,而JNK信号通路的激活与细胞凋亡关系密切,两者还参与多种疾病的病理生理变化过程。JNK信号转导通路与神经退行性病变或其他神经系统相关疾病有着密切联系。在肿瘤的发生以及浸润过程中ERK信号转导通路可能是其主要机制,ERK还参与疼痛的产生,ERK在细胞内的定位可能与神经变性有密切联系。

双氢青蒿素对大鼠烧伤创面愈合及ERK/JNK信号通路的影响

目的探讨双氢青蒿素对大鼠烧伤创面愈合及ERK/JNK信号通路的影响。方法采用金属砝码烧伤法构建大鼠烧伤模型,并将造模成功的大鼠随机分成模型组、双氢青蒿素低(25 mg/kg)、中(50 mg/kg)、高剂量组(100 mg/kg)和磺胺嘧啶银(SS)组,每组10只。另取10只大鼠作为假手术(sham)组,将金属砝码置于37℃温水中,其余按烧伤模型构建相同方法处理大鼠。sham组和模型组使用橄榄油和无水乙醇混悬液涂抹创面,双氢青蒿素低、中、高剂量组使用含双氢青蒿素的橄榄油和无水乙醇混悬液涂抹创面,SS组大鼠使用SS乳膏涂抹。在药物处理3,7,14,21 d时分别观察记录大鼠创面愈合情况并计算愈合率。药物处理21 d时,苏木素-伊红(HE)法检测大鼠创面组织病理变化;ELISA法检测血清中炎症因子水平,包括肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6);试剂盒检测创面组织氧化应激反应因子活性,包括丙二醛(MDA)水平、超氧化物歧化酶(SOD)和过氧化氢酶(CAT);Western blot法检测创面组织ERK/JNK信号通路相关蛋白(ERK1/2、JNK)表达情况。结果模型大鼠出现明显的烧伤创面,说明造模成功。药物处理14,21 d时,与模型组比较,双氢青蒿素低、中、高剂量组和SS组大鼠创面愈合率明显提高(P<0.05)。药物处理21 d时,与sham组比较,模型组大鼠血清TNF-α、IL-1β、IL-6水平及创面组织MDA水平、p-ERK1/2/ERK1/2、p-JNK/JNK明显升高(P<0.05),SOD和CAT活性明显降低(P<0.05)。与模型组比较,双氢青蒿素低、中、高剂量组和SS组大鼠血清TNF-α、IL-1β、IL-6水平及创面组织MDA水平、p-ERK1/2/ERK1/2、p-JNK/JNK明显降低(P<0.05),SOD和CAT活性明显升高(P<0.05)。双氢青蒿素低剂量组与SS组间比较大鼠血清TNF-α、IL-1β、IL-6水平及创面组织MDA水平、SOD和CAT活性、p-ERK1/2/ERK1/2、p-JNK/JNK无明显差异(P>0.05);与SS组比较,双氢青蒿素中剂量组大鼠血清TNF-α水平及创面组织MDA水平、p-ERK1/2/ERK1/2、p-JNK/JNK显著降低(P<0.05),SOD、CAT活性显著升高(P<0.05),而IL-1β、IL-6水平差异不显著(P>0.05);与SS组比较,双氢青蒿素高剂量组大鼠血清TNF-α、IL-1β、IL-6水平及创面组织MDA水平、p-ERK1/2/ERK1/2、p-JNK/JNK显著降低(P<0.05),SOD、CAT活性显著升高(P<0.05)。结论双氢青蒿素对大鼠烧伤创面愈合具有促进作用,可能是通过抑制ERK/JNK信号通路,减轻炎症和氧化应激反应实现的。

锦红片对急性胆源性感染大鼠JNK、P38、ERK mRNA及蛋白表达调控的研究

目的:探讨锦红片对急性胆源性感染大鼠MAPK通路信号分子JNK、P38、ERK mRNA及蛋白表达的影响。方法:40只雄性SD大鼠随机分为假手术组、模型组、锦红片组及庆大霉素组,每组10只。采用胰胆管内注入大肠杆菌法建立急性胆源性感染大鼠模型,假手术组仅开腹关腹;各药物组在造模后第1天予相应剂量药物灌胃,假手术组和模型组予等量生理盐水灌胃。各组均在药物干预后第5天取材,分别用HE染色和电镜观察小肠大体形态及超微结构变化,Real time-PCR和Western blot分别检测小肠JNK、P38、ERK mRNA及蛋白表达。结果:模型组大鼠大体形态和超微结构均有不同程度病理改变,锦红片和庆大霉素能一定程度上改善。模型组大鼠小肠组织JNK、P38及ERK mRNA表达均高于假手术组(P<0.01);锦红片及庆大霉素组大鼠小肠组织JNK、P38及ERK mRNA表达低于模型组(P<0.01);锦红片组大鼠小肠组织ERK mRNA表达低于庆大霉素组(P<0.01)。模型组大鼠小肠组织本底及磷酸化JNK、P38及ERK蛋白表达高于假手术组(P<0.01);锦红片组大鼠小肠组织本底及磷酸化JNK及P38蛋白表达低于模型组(P<0.05或P<0.01),庆大霉素组大鼠小肠组织本底JNK蛋白表达低于模型组(P<0.01);锦红片组大鼠小肠组织本底JNK蛋白表达低于庆大霉素组(P<0.01);锦红片组和庆大霉素组大鼠小肠组织本底及磷酸化ERK蛋白表达与模型组比较,差异均无统计学意义(P>0.05)。结论:"下调MAPK信号通路中JNK、P38及ERK信号分子—抑制肠黏膜上皮细胞过度凋亡—保护肠黏膜屏障—防止炎症二次打击"可能是锦红片治疗急性胆道感染的重要作用途径之一。

The JNK Pathway and Neuronal Migration

The c-Jun N-terminal kinases （JNKs） are important regulators of a variety of physiological and pathological processes both in the central and in the peripheral nervous systems. JNKs are considered as crucial mediators of neuronal cell death in response to stress and injury. However, recent studies have provided substantial evidence that the JNK pathway plays an important role in neuronal migration. Here, we will give a brief introduction of the JNK signaling pathway and put more emphasis on its role in nettronal migration.

芪参益气滴丸对大鼠心肌缺血再灌注损伤的MAPK/p38/JNK/ERK信号通路的研究

目的探讨芪参益气滴丸对大鼠心肌缺血再灌注损伤的MAPK/p38/JNK/ERK信号通路的研究。方法随机将大鼠分为假手术组(Sham)和心肌缺血再灌注组(I/R),后者又分为:模型组(I/R),芪参益气高剂量组(QSYQ-H)和低剂量组(QSYQ-L);血流动力学检测左心室收缩压峰值(LVSP)及左心室内压最大上升速率(+dp/dtmax)、左心室内压最大下降速率(-dp/dtmax);ELISA法检测血浆中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平;qRT-PCR和蛋白质印迹法(Western blotting)检测心肌细胞p38,JNK,ERK mRNA和蛋白的水平。结果与假手术组比较,模型组大鼠LVSP和±dp/dtmax明显较低(P<0.01);与模型组相比,芪参益气滴丸高低剂量组大鼠LVSP和±dp/dtmax均显著升高,且芪参益气滴丸高及低剂量组呈浓度依赖效应,差异有统计学意义(P<0.01);与假手术组比较,模型组大鼠血浆中TNF-α、IL-1β和IL-6蛋白含量明显升高(P<0.01);与模型组相比,芪参益气滴丸高低剂量组大鼠血浆中TNF-α、IL-1β和IL-6蛋白含量明显降低,且芪参益气滴丸高及低剂量组呈浓度依赖效应,差异有统计学意义(P<0.01);与假手术组相比,模型组对大鼠心肌细胞p38、JNK和ERK mRNA和蛋白含量明显降低,差异有统计学意义(P<0.01);与模型组相比,芪参益气滴丸高低剂量组大鼠心肌细胞p38、JNK和ERK mRNA和蛋白含量明显升高,且芪参益气滴丸高及低剂量组呈浓度依赖效应,差异有统计学意义(P<0.01)。结论芪参益气滴丸能减小大鼠心肌缺血再灌注的损伤程度,有可能是通过p38/JNK/ERK激活,抑制炎症相关因子TNF-α、IL-1β和IL-6的表达。

Estrogen up-regulates MMP2/9 expression in endometrial epithelial cell via VEGF-ERK1/2 pathway

Objective:To study the effect of estrogen on anovulatory dysfunctional uterine bleeding(ADUB).Methods:Primary endometrial epithelial cells of Hainan Lizu female was cultured and hydrolylic activity of gelalinase was determined by gelatin zymography analysis.Cellular mRNA and protein synthesis was blocked respectively to determine whether the increased expression of MMP-2/9 was induced by estrogen.The expression of VEGF was blocked by siRNA.After treatment with various factors.MMP-9,VEGF,total Erk and phosphorylated Erk expression in primary uterine epithelial cells was detected by Western blotting analysis.Cell MMP-2/9mRNA levels was measured by real-time RT-PCR.Results:The activity and expression of MMP2/9 was inereased in the endometrium of patients with ADUB.Estrogen could up-regulate the expression of VEGF and activate Erk 1/2-Elk1 signal path.After interference by siRNA,ERK1/2 pathway was blocked in cells,and the expression of MMP-2/9 was down-regulated.ERK1/2 specific blocker U0126 blocked ERK phosphorylation,and it could down-regulate the expression of MMP-2/9.Conclusions:The results showed that the estrogen can increase the expression of VEGF,and thus activate ERK1/2 pathway to induce MMP-2/9 expression.

C-jun N-terminal Kinase-mediated Signaling Is Essential for Staphylococcus Aureus-induced U937 Apoptosis

Objective To investigate the effect of SP600125, a specific c-jun N-terminal protein kinase (JNK) inhibitor, on Staphylococcus aureus (S. aureus)-induced U937 cell death and the underlying mechanism. Methods The human monocytic U937 cells were treated with S. aureus at different time with or without SP600125. Cell apoptosis was analyzed by flow cytometry. JNK, Bax, and caspase-3 activities were detected by Western blotting. Results S. aureus induced apoptosis in cultured U937 cells in a time-dependent manner. Expression of Bax and phospho-JNK significantly increased in S. aureus-treated U937 cells, and the level of activated caspase-3 also increased in a time-dependent manner. Inhibition of JNK with SP600125 significantly inhibited S. aureus-induced apoptosis in U937 cells. Conclusions S. aureus can induce apoptosis in U937 cells by phosphorylation of JNK and activation of Bax and caspase-3. SP600125 protects U937 cells from apoptosis induced by S. aureus via inhibiting the activity of JNK.

苏木提取物对动脉粥样硬化模型ApoE-/-小鼠氧化应激作用及对p38/JNK/ERK的影响

目的:通过建立动脉粥样硬化小鼠模型,观察苏木提取物对氧化应激的影响,并明确与p38/JNK/ERK信号通路的关系,进而探讨苏木提取物抗AS的作用机制。方法:所有小鼠适应性饲养2周后,将40只ApoE-/-小鼠随机分为,模型组、苏木提取物低、中、高剂量组,每组10只;10只相同遗传背景的C57BL/6J小鼠设为空白组,其中空白组小鼠给予普通饲料喂养,其余四组小鼠给予高脂饲料喂养8周进行模型复制,第9周开始灌胃,连续灌胃4周,12周后分离小鼠主动脉根部,制备石蜡切片,油红O染色观察小鼠主动脉窦脂质沉积情况;ELISA法检测小鼠主动脉中MDA、GSH-Px、SOD水平;Western blot法检测小鼠主动脉中p38、JNK、ERK蛋白磷酸化的表达。结果:苏木提取物低、中、高剂量组够降低AS模型小鼠主动脉窦脂质沉积水平;降低小鼠主动脉中MDA水平,升高主动脉中GSH-Px、SOD水平;并可下调小鼠主动脉中p38、JNK、ERK蛋白磷酸化的表达。结论:苏木提取物能够通过抗氧化应激途径发挥抗AS的作用,部分机制可能与抑制p38/JNK/ERK信号通路活化有关。

Protective effects of Yishen Sanjie Huayu compound on the renal artery disease in rats with IgA nephropathy through ERK/NF-κB pathway

Objective:To observe the effect of Yishen Sanjie Huayu compound prescription on ERK/NF-κB signaling pathway in IgA nephropathy(IgAN)rats,and explore its effect on preventing and treating IgA nephropathy intrarenal arteriole disease.Methods:Fifty-five male SD rats were randomly divided into blank group,model group,ShenfukangⅡcapsule group and Losartan potassium tablet group.Bovine serum albumin(BSA)was used for intragastric administration and carbon tetrachloride(CCl4).IgAN rat model was established by subcutaneous injection and lipopolysaccharide(LPS)tail vein injection.ShenfukangⅡcapsule group and Losartan potassium tablet group were given each drug suspension 2ml/head/d one week after modeling Gavage was started.The blank group and the model group were given an equal volume of normal saline.The 24h urine protein(UTP)of the rats was measured at 4,8,and 12 weeks after the administration,and the blood creatinine(SCr)was measured after 12 weeks.,urea nitrogen(BUN),aldosterone(ADS),angiotensinⅡ(AngⅡ),immunohistochemical Envi-sion System two-step method to detect vascular endothelial growth factor(VEGF)and human matrix in the whole rat kidney and small artery area The expression of metalloproteinase-9(MMP-9),proliferating cell nuclear antigen(PCNA),extracellular regulatory protein kinase(ERK)1/2,nuclear transcription factor-κB(NF-κB),and the small arteries of rat kidney tissue The intima,media,vessel wall/vascular outer diameter value.Results:Compared with the model group,the expressions of VEGF,MMP-9,PCNA,ERK1/2 and NF-κB in kidney tissues of the ShenfukangⅡcapsule group and the Losartan potassium tablet group decreased(P<0.05),24hUTP and SCr,BUN level decreased(P<0.05),kidney tissue damage was alleviated;intima and vessel wall/vascular outer diameter values were significantly reduced(P<0.01),there was no significant difference in ADS between the groups.The AngⅡof the Tanpotassium tablets group was lower than that of the model group(P<0.05).Conclusion:Yishen Sanjie Huayu compound can inhibit the ERK/NF-κB signaling pathway in rats with IgA nephropathy,reduce the levels of VEGF,MMP-9,PCNA,ERK1/2,NF-κB,and inhibit intrarenal arteriole vascular endothelial cells Proliferate and reduce kidney damage.