水飞蓟素通过共同抑制TLR4/NF-κB和TNF-α/ROS/P38MAPK通路减轻糖尿病肾病大鼠肾脏损伤的研究

目的:探讨水飞蓟素通过共同抑制Toll样受体4/核因子-κB(TLR4/NF-κB)和肿瘤坏死因子α/活性氧簇/P38丝裂原活化蛋白激酶(TNF-α/ROS/P38MAPK)通路减轻糖尿病肾病(DN)大鼠肾脏损伤的机制。方法:选取健康SD大鼠50只,按随机数字表法分为对照组、模型组、低剂量水飞蓟素组、中剂量水飞蓟素组、高剂量水飞蓟素组,每组10只。建模成功并治疗8周后,生化分析仪测定各组大鼠空腹血糖(FBG)、血肌酐(Scr)、尿素氮(BUN)、胱抑素C(Cys-C)、β_(2)微球蛋白(β_(2)-MG)、24 h尿蛋白(UTP)水平;计算肾脏指数;试剂盒检测各组大鼠肾组织丙二醛(MDA)、总抗氧化能力(T-AOC)水平;RT-PCR检测各组大鼠TLR4,NF-κB,TNF-α和P38MAPK等mRNA的表达水平。结果:与对照组相比,模型组体质量、T-AOC均降低(P<0.05),肾脏指数和FBG,Scr,BUN,Hcy,Cys-C,β_(2)-MG,24 h UTP,MDA,TLR4 mRNA,NF-κB p65 mRNA,TNF-αmRNA,P38MAPK mRNA均升高(P<0.05)。低、中、高剂量水飞蓟素组体质量、T-AOC水平均低于对照组,且均高于模型组(P<0.05);低、中、高剂量水飞蓟素组肾脏指数和FBG,Hcy,Cys-C,β_(2)-MG,24 h UTP,MDA,TLR4 mRNA,NF-κB p65 mRNA,TNF-αmRNA,P38MAPK mRNA均高于对照组,且均低于模型组(P<0.05);低、中剂量水飞蓟素组Scr,BUN水平均高于对照组,且低于模型组(P<0.05);高剂量水飞蓟素组Scr,BUN水平均低于模型组(P<0.05),高于对照组,但差异无统计学意义(P>0.05)。结论:水飞蓟素可调控TLR4/NF-κB和TNF-α/ROS/P38MAPK通路,可通过抑制其激活减轻氧化应激、减轻DN大鼠的肾脏损伤。

Betulinic acid protects against ovarian impairment by decreasing F-2 toxin-induced oxidative stress and inflammation associated with the downregulation of p38 expression in mice

F-2 toxin is an estrogenic mycotoxin that causes reproductive disorders in animals.Betulinic acid(BA)is a natural pentacyclic lupane-structure triterpenoid that has diverse pharmacological activities.In this study,the antioxidative and anti-inflammatory effects of BA and its underlying mechanism are explored in F-2 toxin-triggered mouse ovarian damage.We found that BA alleviated the F-2 toxin-induced ovarian impairment by stimulating follicle growth,reducing inflammatory cell infiltration,repairing damaged mitochondria and endoplasmic reticulum.Simultaneously,BA not only reversed F-2 toxin-induced reduction of follicle stimulating hormone(FSH)and luteinizing hormone(LH)levels in the serum,but also restrained the protein expression of the estrogen receptors a(ERa)and ERβ.Moreover,BA restored the balance of F-2 toxin-induced ovarian redox system disorders.Subsequently,we found that 0.25 mg/kg BA played an anti-inflammatory role in the F-2 toxin-induced ovarian impairment by decreasing interleukin-1β(IL-1β).IL-6,and tumor necrosis factor-α(TNF-α)mRNA expression,as well as inhibiting p38 protein expression.These data demonstrated that BA exerts its protective effect on F-2 toxin-induced ovarian oxidative impairment and inflammation by inhibiting p38 expression,which implies a natural product-based medicine to ameliorate F-2 toxin-caused female reproductive toxicity and provides a detoxifying method for food contaminated by mycotoxin.

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.

Promotion effect of FOXCUT as a microRNA sponge for miR-24-3p on progression in triple-negative breast cancer through the p38 MAPK signaling pathway

Background:Triple-negative breast cancer(TNBC)is a type of highly invasive breast cancer with a poor prognosis.According to new research,long noncoding RNAs(lncRNAs)play a significant role in the progression of cancer.Although the role of lncRNAs in breast cancer has been well reported,few studies have focused on TNBC.This study aimed to explore the biological function and clinical significance of forkhead box C1 promoter upstream transcript(FOXCUT)in triple-negative breast cancer.Methods:Based on a bioinformatic analysis of the cancer genome atlas(TCGA)database,we detected that the lncRNA FOXCUT was overexpressed in TNBC tissues,which was further validated in an external cohort of tissues from the General Surgery Department of the First Affiliated Hospital of Nanjing Medical University.The functions of FOXCUT in proliferation,migration,and invasion were detected in vitro or in vivo.Luciferase assays and RNA immunoprecipitation(RIP)were performed to reveal that FOXCUT acted as a competitive endogenous RNA(ceRNA)for the microRNA miR-24-3p and consequently inhibited the degradation of p38.Results:lncRNA FOXCUT was markedly highly expressed in breast cancer,which was associated with poor prognosis in some cases.Knockdown of FOXCUT significantly inhibited cancer growth and metastasis in vitro or in vivo.Mechanistically,FOXCUT competitively bounded to miR-24-3p to prevent the degradation of p38,which might act as an oncogene in breast cancer.Conclusion:Collectively,this research revealed a novel FOXCUT/miR-24-3p/p38 axis that affected breast cancer progression and suggested that the lncRNA FOXCUT could be a diagnostic marker and therapeutic target for breast cancer.

Mannitol inhibits the proliferation of neural stem cell by a p38 mitogen-activated protein kinase-dependent signaling pathway

Purpose:Mannitol is one of the first-line drugs for reducing cerebral edema through increasing the extracellular osmotic pressure.However,long-term administration of mannitol in the treatment of cerebral edema triggers damage to neurons and astrocytes.Given that neural stem cell(NSC)is a subpopulation of main regenerative cells in the central nervous system after injury,the effect of mannitol on NSC is still elusive.The present study aims to elucidate the role of mannitol in NSC proliferation.Methods:C57 mice were derived from the animal house of Zunyi Medical University.A total of 15 pregnant mice were employed for the purpose of isolating NSCs in this investigation.Initially,mouse primary NSCs were isolated from the embryonic cortex of mice and subsequently identified through immunofluorescence staining.In order to investigate the impact of mannitol on NSC proliferation,both cell counting kit-8 assays and neurospheres formation assays were conducted.Thein vitro effects of mannitol were examined at various doses and time points.In order to elucidate the role of Aquaporin 4(AQP4)in the suppressive effect of mannitol on NSC proliferation,various assays including reverse transcription polymerase chain reaction,western blotting,and immunocytochemistry were conducted on control and mannitol-treated groups.Additionally,the phosphorylated p38(p-p38)was examined to explore the potential mechanism underlying the inhibitory effect of mannitol on NSC proliferation.Finally,to further confirm the involvement of the p38 mitogen-activated protein kinase-dependent(MAPK)signaling pathway in the observed inhibition of NSC proliferation by mannitol,SB203580 was employed.All data were analyzed using SPSS 20.0 software(SPSS,Inc.,Chicago,IL).The statistical analysis among multiple comparisons was performed using one-way analysis of variance(ANOVA),followed by Turkey’’s post hoc test in case of the data following a normal distribution using a Shapiro-Wilk normality test.Comparisons between 2 groups were determined using Student’s t-test,if the data exhibited a normal distribution using a Shapiro-Wilk normality test.Meanwhile,data were shown as median and interquartile range and analyzed using the Mann-WhitneyU test,if the data failed the normality test.A p<0.05 was considered as significant difference.Results:Primary NSC were isolated from the mice,and the characteristics were identified using immunostaining analysis.Thereafter,the results indicated that mannitol held the capability of inhibiting NSC proliferation in a dose-dependent and time-dependent manner using cell counting kit-8,neurospheres formation,and immunostaining of Nestin and Ki67 assays.During the process of mannitol suppressing NSC proliferation,the expression of AQP4 mRNA and protein was downregulated,while the gene expression of p-p38 was elevated by reverse transcription polymerase chain reaction,immunostaining,and western blotting assays.Subsequently,the administration of SB203580,one of the p38 MAPK signaling pathway inhibitors,partially abrogated this inhibitory effect resulting from mannitol,supporting the fact that the p38 MAPK signaling pathway participated in curbing NSC proliferation induced by mannitol.Conclusions:Mannitol inhibits NSC proliferation through downregulating AQP4,while upregulating the expression of p-p38 MAPK.

Polysaccharides from Agrocybe cylindracea residue alleviate type 2-diabetes-induced liver and colon injuries by p38 MAPK signaling pathway

In this experiment,we investigated the possible mechanism of polysaccharides from Agrocybe cylindracea residue (PACR) on ameliorating the type-2-diabetes-induced liver and colon injuries.Animal experiments have proved that PACR could reduce the oxidative damage and inflammatory response.Meanwhile,the PACR could restore lipid levels,decrease the level of liver and colon lesions in injured mice,and finally play a role in protecting liver and colon.The results showed that PACR could be used as a supplement to decrease blood glucose and relieve T2DM and reduce oxidative stress and inflammatory response by inhibiting the activation of p38 MAPK signaling pathway.

The functional analysis of transiently upregulated miR-101 suggests a “braking” regulatory mechanism during myogenesis

Skeletal muscle differentiation is a highly coordinated process that involves many cellular signaling pathways and microRNAs(miRNAs).A group of muscle-specific miRNAs has been reported to promote myogenesis by suppressing key signaling pathways for cell growth.However,the functional role and regulatory mechanism of most non-muscle-specific miRNAs with stage-specific changes during differentiation are largely unclear.Here,we describe the functional characterization of miR-101a/b,a pair of non-muscle-specific miRNAs that show the largest change among a group of transiently upregulated miRNAs during myogenesis in C2C12 cells.The overexpression of miR-101a/b inhibits myoblast differentiation by suppressing the p38/MAPK,Interferon Gamma,and Wnt pathways and enhancing the C/EBP pathway.Mef2a,a key protein in the p38/MAPK pathway,was identified as a direct target of miR-101a/b.Interestingly,we found that the long non-coding RNA(lncRNA)Malat1,which promotes muscle differentiation,interacts with miR-101a/b,and this interaction competes with Mef2a mRNA to relieve the inhibition of the p38/MAPK pathway during myogenesis.These results uncovered a“braking”role in differentiation of transiently upregulated miRNAs and provided new insights into the competing endogenous RNA(ceRNA)regulatory mechanism in myoblast differentiation and myogenesis.