Effect of aqueous extract of Semen Cassiae on endogenous metabolomics in urine of rats北大核心CSCD

OBJECTIVE: To investigate the effect of aqueous extract of Semen Cassiae (AESC) on endogenous metabolites in urine of rats by metabolomics based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) to reveal the possible ways of metabolism and mechanism of action in rats caused by AESC. METHODS: Twsenty-eight male Sprague-Dawley (SD) rats were randomly equally divided into 4 groups: such normal control group, AESC 1.5, 5 and 15 g·kg-1 groups. After intragastric administration for 14 d, the urine was collected with metabolic cages. The urine metabolic profiling was analyzed using UPLC-QTOF-MS, based on which the principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) models were established for metabolomic analysis. Potential biomarkers were screened using variable importance in the projection (VIP) and t test. RESULTS: The results of PCA showed that samples of each group were clustered, all the groups were separated, and that the distance between AESC groups and normal control group was increased in a dose-dependent manner. The relative content of proline betaine and uric acid were 18.4±2.3 and 15.7±2.0, 16.3±4.5 and 14.7±3.0 in the AESC 5 and 15 g· kg-1 groups, significantly lower than that of the normal control group, which was 25.0±3.4 and 29.0±4.8(P<0.01), but that of AESC 1.5 g · kg-1 group did not statistically differ from that of normal control group. In AESC 1.5, 5 and 15 g·kg-1 groups, the relative content of glycine and taurine was 10.0±1.4 and 8.0±1.4, 3.6±0.7 and 66.5±7.3, 45.8±23.6 and 23.0±9.8, which was significantly lower than that of the normal control group, which was 14.6±1.9 and 102.5±25.8(P<0.01). The relative content of 1,7-dimethylguanosine was 4.5±1.2 and 4.6±0.1 in AESC 1.5 and 15 g·kg-1 groups, significantly lower than that of the normal control group, which was 6.5±0.8(P<0.05), but AESC 5 g·kg-1 group did not statistically differ from normal control group. The relative content of citric acid was 26.6±6.3 in the AESC 15 g·kg-1 group, significantly lower than that of the normal control group, which was 67±14(P< 0.01). The relative content of citric acid was 104+20 in the AESC 1.5 g·kg-1 group, significantly higher than that of the normal control group (P<0.01), but AESC 5g·kg-1 group did not statistically differ from normal control group. CONCLUSION: AESC can remarkably change endogenous metabolites and mainly affect the pathways of taurine,purine, amino acid and energy metabolism.     

Effect of total flavonoids of Rosa rugosa on PI3K/AKT pathway and endoplasmic reticulum stress apoptosis in rats with cerebral ischemia-reperfusion injury北大核心CSCD

Aim To investigate the effects of total flavonoids from Rosa rugosa (TFR) on cerebral ischemia reperfusion injury (CIRI) in rats, and to investigate whether TFR inhibited neuronal apoptosis by regulating phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) signaling pathway and endoplasmic reticulum stress (ERS) pathways. Methods SD rats were randomly divided into sham operation group, model group, low-dose group (50 mg · kg -1 · d -1), medium-does group (100 mg · kg -1 · d -1), and high-does group (200 mg· kg -1 · d -1). The injury model of middle cerebral artery occlusion/reperfusion (MCAO/R) was prepared following suture method. Neurobehavioral changes, cerebral infarct size and brain tissue water content were detected 24 h after surgery. HE and Nissl staining were performed to observe pathological indicators. TUNEL staining was used to detect the apoptosis of ischemic nerve cells in brain. Western blot was used to detect the protein levels of Bcl-2, Bax, and cleaved Caspase-3, PI3K, p-PI3K, AKT, p-AKT, GRP78, CHOP and Caspase-12. Results Compared with MCAO/R group, the rats in medium-dose group and high-dose group showed improvement in the neurobehavioral function, decrease in the cerebral infarction area and the degree of cerebral edema, and reduction of the pathological damage of cerebral cortex. Moreover, there was a significantly decrease in the apoptosis rate of nerve cells in medium-dose group and high-dose group. The expression of anti-apoptotic protein Bcl-2 increased, and the pro-apoptotic protein Bax and cleaved-Caspase-3 decreased, the expression of pPI3K/PI3K, p-Akt/AKT increased, and the expression of ERS-related protein GRP78, CHOP, Caspase-12 decreased. Conclusions TFR can inhibit neuronal apoptosis by regulating the PI3K/AKT signaling pathway and ERS pathway, thus playing a protective role in CIRI rats. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Effect of total flavonoids of dracocephalum heterophyllum benth on norepinephrine-induced cardiomyocyte hypertrophy in rats北大核心CSCD

OBJECTIVE: To study the effect of total flavonoids of Dracocephalum heterophyllum (TFDH), a Uygur medicine, on cardiomyocyte hypertrophy induced by norepinephrine (NE), and to provide insights into the mechanism. METHODS: Cardiomyocytes of primarily cultured neonatal rats were used as models. Myocardial hypertrophy was induced by NE 2 umol · L-1. The cells were divided into cell control group, NE 2 umol-L-1 model group, and TFDH 10, 25 and 50 umol-L-1+NE 2 umol-L-1 groups. CCK-8 method was used to observe the activity of myocardial cells, while RT-PCR technique was used to detect the expression of mRNA of cardiac hypertrophy gene atrial natriuretic peptide (ANP) and β-myosin heavy chain (β-MHC). The internal factors were glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Confocal laser scanning was used to detect the surface area of myocardial cells and [Ca2+]. The activity of Ca2+-ATP was measured with enzymatic reaction of fragmentation cells. The concentration of NO and the activity of NOS were determined with colorimetry. RESULTS: Compared with the cell control group, cardiomyocytes were stimulated at 48 h by NE 2 umol · L-1 which could decrease the survival rate of cardiomyocytes from (95±1)% to (78+5)%, surface area increased from (178±29) μm2 to (274±38) μ2 (P<0.05), the expression of mRNA of ANP and β-MHC increased from (1.00±0.01) and (1.00±0.02) to (2.76±0.55) and (2.69±0.31), respectively (P<0.05). The concentration of [Ca2+], increased from 1.00±0.12 to (1.52 ± 0.41) umol-L-1, while the activity of Ca2+-ATP decreased from 1.01±0.14 to (0.41±0.06) umol-L-1 (P<0.05). The concentration of NO decreased from 1.50±0.14 to (1.12±0.05) umol-L-1, and the activity of NOS decreased from 0.86±0.06 to (0.52±0.10) μmol-L-1 (P<0.05). TFDH 10, 25 and 50 μmol · L-1 could inhibit the decline of the survival rate, increase of the surface area and the increased expressions of mRNA of ANP and β-MHC that were induced by NE (P<0.05). At the same time, it also could inhibit the increase of the concentration of [Ca2+], the decreased of activity of Ca2+-ATP, and the decline of the concentration of NO and the activity of NOS (P<0.05). CONCLUSION: TFDH can improve the activity of cardiomyocyte hypertrophy induced by NE, down-regutate mRNA expressions of proto oncogene ANP and β-MHC, and reduce the surface area of cardiomyocytes induced by NE. The mechanism may be related to promoting the release of NO and regulating the concentration of [Ca2+], and the activity of Ca2+-ATP. © 2017 Chinese Journal of Pharmacology and Toxicology. All rights reserved.     

Effect of γ-ray on metabolic enzyme CYP3A1 in rat liver on multiple levels北大核心CSCD

Aim To explore the effect of γ-ray on the mRNA,protein expression levels and metabolic activity level of the key drug metabolic enzyme CYP3A1 in rat liver. Methods Wistar rats were randomly divided into control group, 24 h post-radiation group and 72 h post-radiation group. The experimental group was exposed to total body irradiation of single 6 Gy γ-ray. Blood was collected from the orbital venous plexus for blood routine examination and biochemical analysis 24 h and 72 h after irradiation, and liver tissue was prepared for quantifying expression of CYP3A1 mRNA and liver-specific microRNA (miR-122-5p) through RT-PCR. The expression level of CYP3A1 protein was analyzed by Western blot, and the metabolic activity level of CYP3A1 detected by the specific substrate midazolam combined with LC-MS method. Results Com¬pared with the control group, the weights of the rats in the radiation group significantly decreased, and the number of white blood cells was markedly reduced. Simultaneously, the activities of alanine aminotrans-ferase and alkaline phosphatase continuously descended, as well as the levels of total bilirubin and bile acid significantly increased, which indicated that the liver may be damaged after radiation. The relative expression of CYP3A1 mRNA continued to increase significantly 24 h and 72 h after irradiation. CYP3A1 protein expression and metabolic activity levels showed an obvious increasing trend 24 h after irradiation, and rose significantly 72 h after irradiation compared with the control group. At the same time, the expression of miR-122-5p in liver of rats in the 24 h and 72 h post-radiation group continued to decrease rapidly compared with the control group. Conclusions γ-ray radiation may arouse damage effect on liver, which leads to the continuous up-regulation of the mRNA and protein expression levels of the capital metabolic enzyme CYP3A1 in liver tissue, as well as the elevation of the metabolic activity level. The regulatory mechanism might be related to miR-122-5p. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Effect of corilagin on cholesterol metabolism in macrophages and its mechanism北大核心CSCD

Aim To elucidate the effect of corilagin (Cor) on cholesterol metabolism in macrophages and the underlying mechanism. Methods Molecular docking was applied to predict the protein target of Cor on cellular cholesterol metabolism. The RAW264.7 macrophage foam model induced by 80 mg • L-1 oxidized low density lipoprotein (ox-LDL) was established to evaluate the activity of Cor on lowering-cholesterol. The expression of genes and proteins related with cholesterol metabolism were detected by q-PCR and Western blotting,respectively. Then the activity of Cor on lipid metabolism was validated in ApoE mice fed with high-fat-diet. Results Cor and Class A Scavenger receptor (SRA), CD36, peroxisome proliferator-activated receptor γ (PPAR-γ), ATP binding cassette transporter Gl(ABCGl), which associated with cholesterol metabolism, could form hydrogen bonds and hydrophobic interactions. Cell experiments showed that Cor (60,120 and 240 μmol • L-1) significantly decreased TC content in macrophages, Cor could down-regulate SRA and CD36 gene expression, SRA protein expression, up-regulate the expression of ABCA1 and ABCG1 genes. Animal experiments demonstrated that Cor (15,30 and 60 mg • k g - 1 ) could decrease the serum TMAO content, the plaque area and formation of foam cells in the aortic root,the expression levels of CD36 and SRA fluorescent proteins in aortic root plaques. Conclusions Cor could inhibit the formation of macrophage foam cells through the regulation of cholesterol metabolism mediated by CD36,SRA, ABCA1 and ABCG1 to cure the AS. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Establishment and characteristic analysis of fibroblast-like synoviocytes in rats with adjuvant arthritis北大核心CSCD

Aims To establish the methods of primary culture of fibroblast-like synoviocytes in rats with adjuvant arthritis (AA-FLS) and analyze the feature and to investigate the possibility of AA-FLS as the model for the RA in vitro. Methods The synovial cells obtained from the SD rats were immunized by the Mtb and identified by morphology and immunocytochemistry. The viability of AA-FLS was assessed by Cell Counting Kit-8. ELISA was applied to detect TNF-α and IL-lβ in cell media. Apoptosis was measured by Hochest 33258. The expressions of mitochondrial apoptosis-re-lated molecules, including Bcl-2, Bax, Pro-caspase-3 and Cleave-caspase-3 were determined by Western Blot. Result In isolated primary synovial cells, more than 95% of AA-FLSs were fusiform. Immunocyto-chemistry result showed a positive expression of vimen-tin and a negative expression of CD68 in AA-FLS. Cell proliferation of AA-FLS was higher than FLS and cell apoptosis of AA-FLS was curbed. Western blot data demonstrated that the protein expressions of Bcl-2, Bax were regulated and the expression of caspase-3 was activated in AA-FLS. Conclusions AA-FLS is biologically characterized by high level proliferation activity and inflammatory cytokines and apoptosis suppression. AA-FLS can be used as the model for the RA in vitro.     

Effect of yeast and potassium oxonate on formation and excretion of uric acid and renal function of SD rats北大核心CSCD

OBJECTIVE: To investigate the effect of potassium oxonate and yeast on formation and excretion of uric acid (UA) and renal function in rats, and to describe the characteristics of hyperuricemia in rats induced by potassium oxonate and yeast. METHODS: SD rats were given yeast 21 g·kg-1 and potassium oxonate 100, 200 and 300 mg·kg-1, respectively, once a day for 35 d. Levels of UA, creatinine (CRE), urea nitrogen (BUN) and the activity of adenosine deaminase (ADA), xanthine oxidase (XOD), guanine deaminase (GuDa) in serum were determined on the 14th 28th and 35th day, respectively. Levels of UA, CRE, BUN, protein in urine, urine specific gravity and urine volume were determined on the 28th day while UA excretion and clearance were counted. These rats were sacrificed on the 35th day to weigh the right kidney and calculate the kidney index. RESULTS: Compared with normal control group, serum UA levels of three treatment groups were significantly higher from the 14th to the 35th day(P<0.01); CRE levels of three treatment groups were significantly higher on the 35th day(P<0.05, P<0.01); ADA activity was significantly lower in the groups of yeast 21 g·kg-1 and potassium oxonate 100 and 300 mg·kg-1 on the 14th day (P<0.01) and in the group of yeast 21 g · kg-1 and potassium oxonate 200 mg · kg-1 on the 14th to 28th day (P<0.05); XOD activity of the groups of yeast 21 g · kg-1 and potassium oxonate 300 mg · kg-1 was significantly higher on the 14th to 28th day (P<0.05, P<0.01); UA excretion and clearance of three treatment groups were significantly lower on the 28th day (P<0.05, P<0.01); the kidney index of the three treatment groups was significantly higher on the 35th day (P<0.01). CONCLUSION: Yeast and potassium oxonate can elevate levels of UA in rats, which may be related to the changes of XOD and ADA activity and the disturbance of UA excretion that is likely associated with kidney damage.     

Effect of HgCl2 on zebrafish embryonic development and nuerexin2a gene expression北大核心CSCD

OBJECTIVE: To investigate the developmental toxicology of mercury chloride (HgCl2) in zebrafish embryos and larvae, and determine its effects on neurobehavior and neurexin (nrxn)2a gene expression. METHODS: Wild-type zebrafish embryos/larvae were exposed to HgCl2 0.2-0.8 μmol · L-1. The mortality, hatching rate and malformation of zebrafish were determined. The content of Hg in zebrafish was detected by inductively coupled plasma mass spectrometry (ICP-MS) analysis. Locomotor activity of zebrafish larvae was detected by a Video-Track system. Cell apoptosis in embryos was observed by the staining of acridine orange (AO). Expression of nrxn2a was examined by in situ hybridization and quantitative PCR(qPCR). RESULTS: Compared with control, mortality started to increase after the zebrafish was treated with HgCl20.8 μmol·L-1 for 72 h and reached 94% at 120 h post fertilization (hpf) (P<0.01). Moreover, the hatching rate was significantly increased in the zebrafish treated with HgCl20.4 μmol·L-1 for 48 h. HgCl2 had significantly deleterious effects on zebrafish embryonic development, as mainly evidenced by spinal curvature and yolk edema. Malformation and Hg accumulation were markedly increased (P<0.05, P<0.01). The content of Hg reached 77 μg·g-1 in zebrafish treated with HgCl20.8 umol · L-1. Moreover, compared with control, the velocity and the proportion of activity were significantly inhibited, while the apoptosis was markedly increased in zebrafish larvae exposed with HgCl20.4 umol · L-1 (P<0.05, P<0.01). mRNA level of nrxn2aa was up-regulated in zebrafish embryos at 24 hpf (P<0.01) and 72 hpf (P<0.05), while mRNA level of nrxn2ab was down-regulated at 24 and 48 hpf (P<0.05). CONCLUSION: The development of zebrafish embryos and larvae is inhibited because of toxicity of HgCl2, which is probably associated with changes in nrxn2a expression.     

Cytotoxicity and mitochondrial injury mechanism of matrine on PC12 cells北大核心CSCD

OBJECTIVE: To study the toxicological effect of matrine (MT) on PC12 cells and the mechanism of mitochondrial damage. METHODS: After treatment with MT 2, 4 and 8 mmol · L-1 for 8, 16, 24 and 48 h, respectively, the viability of PC12 cells was measured with methylthiazolyltetrazolium assay. PC12 cells were treated with MT 2, 4 and 8 mmol · L-1 for 24 h, before the morphological changes were observed by Hoechst33342 staining. The superoxide dismutase (SOD) activity and malondialde-hyde (MDA) content in cells were detected using hydroxylamine method and thiobarbituric acid method, apoptosis rate and reactive oxygen species (ROS) of PC 12 cells were detected with flow cytometry, mitochondrial membrane potential (MMP) was detected with JC-1 staining, and the expressions of procaspase 3, procaspase 9, cleaved-caspase 3, Bax and Bcl-2 were detected with Western blotting. RESULTS: MT inhibited the growth of PC12 cells in a time- and concentration-dependent manner. After being treated with MT for 24 h, the nuclei of PC12 cells in MT groups showed chromatin agglutination and partial rupture to different degrees, and compared with normal control group, the apoptosis rates of MT 2, 4 and 8 mmol. L-1 groups were significantly increased (P<0.01). Intracellular ROS and MDA levels increased (P<0.05, P<0.01), SOD activity decreased (P<0.01), and MMP decreased. The expressions of procaspase 9, procaspase 3 and Bcl-2 were significantly decreased (P<0.01, P<0.05), the expressions of cleaved-caspase 3 and Bax were significantly increased (P<0.05, P<0.01), and the ratio of Bcl-2/Bax was significantly decreased (P<0.01). CONCLUSION: MT has toxic effect on PC12 cells and induces apoptosis through ROS mediated mitochondrial pathway. © 2017 Chinese Journal of Pharmacology and Toxicology. All rights reserved.     

The effects of ethanol,estrogen, and hexachlorobenzene on the activities of hepatic δ-aminolevulinate synthetase, δ-aminolevulinate dehydratase,and uroporphyrinogen decarboxylase in male rats

To determine if clinically observed disorders in heme biosynthetic enzymes, known as sporadic porphyria cutanea tarda (PCT), could be reproduced in experimental animals, male Fischer rats were treated with ethanol, estrogen and hexachlorobenzene (HCB). A series of heme biosynthetic enzymes were assayed. In the rats given free access to 8% ethanol-drinking water for 15 weeks, -aminolevulinate (ALA) dehydratase was significantly reduced in erythrocytes. In the liver, ALA synthetase and uroporphyrinogen (UROgen) decarboxylase activities remained unchanged. In bone marrow cells, these activities did not change markedly. In the rats treated with estrogen (1 mg estrioltripropionate /rat/week, IM), no body weight gain was observed during the treatment for 15 weeks and urinary ALA excretion increased to 1.7 fold over normal level. In the liver, a significant increase was observed in the activity of ALA dehydratase, but other enzymes remained within the normal level. In bone marrow cells and erythrocytes, ALA dehydratase was also increased. ALA synthetase increased only in bone marrow cells to 2.1 times higher than the control level. In rats fed 0.3% HCB-diet for 8 weeks, urinary excretion of ALA, coproporphyrin and uroporphyrin increased to 2.4, 3.3 and 3.8 times higher than the controls, respectively. In the liver, an increase was observed in ALA synthetase, while a decrease was observed in ALA dehydratase and UROgen decarboxylase. In bone marrow cells and erythrocytes, ALA dehydratase was reduced and activities of other enzymes did not show any changes.These results indicate that alcohol, estrogen and HCB do not produce phenomena similar to those observed clinically in PCT.     

Effect of catecholamic acid on detoxication and distribution of NiCl_2 in mice and rats

目的：研究双酚胺酸（ＣＢＭＩＤＡ）对氯化镍的解毒作用．方法：ＮｉＣｌ２中毒后，立即给予ＣＢＭＩＤＡ，记录动物存活数；小鼠ｉｖ６３ＮｉＣｌ２后给药，测定２４ｈ组织中６３镍；用整体放射自显影术，显示小鼠体内６３镍分布．结果：ｓｃＣＢＭＩＤＡ０５－１５ｇ·ｋｇ－１对ｉｐＮｉＣｌ２５００ｍｇ·ｋｇ－１有解毒作用；小鼠ｉｐＮｉＣｌ２ＬＤ５０为８２８ｍｇ·ｋｇ－１，给药１５或２５ｇ·ｋｇ－１，ＬＤ５０分别为７８９和８２０ｍｇ·ｋｇ－１；大鼠ｉｍＣＢＭＩＤＡ５００ｍｇ·ｋｇ－１使ＮｉＣｌ２的ＬＤ５０提高８倍；组织中６３镍测定和定位显示，ＣＢＭＩＤＡ减少肺和血液中６３镍，增加了骨中６３镍，２４ｈ尿、粪６３镍排出与对照组无明显差异．结论：ＣＢＭＩＤＡ有效地解除镍毒性，提高动物存活率，降低镍在肺部的滞留．     

Effects of Weichang'an Pill on contraction of isolated ileum smooth muscles in rats北大核心CSCD

目的研究胃肠安丸(Weichang′an pill,WCA)、胃肠安丸醇提物(Ethanol extract of Weichang′an pill,EE)、胃肠安丸水提物(Water extract of Weichang′an pill,WE)及其活性成分对乙酰胆碱(Acetylcholine,ACh)诱导的大鼠离体回肠平滑肌收缩的影响及机制。方法采用离体组织浴实验,在ACh的作用下,加入WCA、EE、WE或其活性成分,记录离体大鼠回肠平滑肌的收缩张力;通过分子对接的方法探究活性成分与毒蕈碱型乙酰胆碱M3受体的结合亲和力。结果WCA、EE、WE均可明显抑制ACh诱导的回肠平滑肌兴奋性收缩。活性成分木香烃内酯、去氢木香内酯、檀香醇、麝香酮、大黄素、大黄酚、大黄素甲醚、巴豆苷、厚朴酚及和厚朴酚也对ACh诱导的回肠平滑肌收缩有明显的抑制作用。结论WCA、EE、WE及其活性成分可能通过阻断回肠平滑肌细胞膜上的M3受体与ACh的结合,发挥促进肠平滑肌松弛的作用。     

Effects of dagliflozin on atrial tachyarrhythmia in rats with pulmonary arterial hypertension related right heart failure and its mechanisms北大核心CSCD

目的探讨达格列净(dapagliflozin,DAPA)对肺动脉高压(pulmonary arterial hypertension,PAH)致右心衰竭(right heart failure,RHF)大鼠房性心律失常(atrial tachyarrhythmia,AT)的影响。方法60只♂SD大鼠随机分为4组:对照组(CTL组)、模型组(MCT组)、MCT+低剂量DAPA干预组(MCT+LD组)和MCT+高剂量DAPA干预组(MCT+HD组),持续干预35 d后,完成模型及心功能评价,心房结构重构评估,炎症因子检测,在体心脏电生理实验。结果DAPA可降低模型大鼠的平均肺动脉压(PAP)及平均右心室压(mRVP)(P<0.05),减轻炎症反应(P<0.05),减轻右心房纤维化(P<0.05),降低AT诱发率(P<0.05)及平均AT持续时间(mean atrial tachyarrhythmia duration,MATD)(P<0.05),其程度在高剂量DAPA干预组中更明显。结论DAPA能够降低PAH致RHF大鼠的AT易感性,其机制主要可能与DAPA抑制系统炎症和抗心房纤维化有关。     

Effect of inhibition of γ-glutamyltranspeptidase by AT-125 (Acivicin) on glutathione and cysteine levels in rat brain and plasma

Summary AT-125 (Acivicin) is an inhibitor of -glutamyltranspeptidase (-GTP) which initiates glutathione catabolism to cysteine. We measured plasma and brain glutathione and cysteine in rats treated with AT-125. Six h after AT-125 treatment, plasma glutathione had increased 6-fold and plasma cysteine had fallen significantly. Brain cysteine fell after 24 h of AT-125 treatment, and brain glutathione had also decreased 18%. AT-125 pretreatment inhibited brain uptake of ³⁵S when it was given as ³⁵S-GSH but had no effect when it was given as ³⁵S-cysteine. These results suggest that plasma glutathione is catabolized by -GTP, and cysteine derived from it is taken up by the brain. N-acetylcysteine was administered to AT-125 treated rats in an attempt to supply cysteine to the brain in the face of -GTP inhibition. N-acetylcysteine supported brain glutathione levels, suggesting that it can serve as a source of cysteine under these conditions.Abbreviations -GTP -glutamyltranspeptidase - GSH reduced glutathione - TCA trichloroacetic acid     

Effect of inorganic active element on hypoxia inducing factor-1α and cytokines in chronic ulcer tissue in rats and its action mechanism

目的 检测无机活性元素(德莫林)对大鼠皮肤慢性溃疡组织的影响,并对其作用机制进行分析.方法 制备SD大鼠慢性溃疡模型,实验组应用无机活性元素,对照组常规消毒换药,另设空白组.观察3组溃疡的愈合时间.应用RT-PCR技术对3组溃疡组织乏氧诱导因子(HIF)-1α、表皮细胞生长因子(EGF)、碱性成纤维细胞因子(bFGF)、血管内皮细胞生长因子(VEGF)的表达进行检测,并对其差异进行比较分析.结果 与对照组比较,实验组创面面积缩小更为明显(P＜0.05);实验组、对照组的HIF-1α、EGF、bFGF和VEGF表达均明显高于正常皮肤(P＜0.05),实验组EGF、bFGF和VEGF表达高于对照组,而HIF-1α表达低于对照组(P＜0.05).结论 无机活性元素对慢性溃疡的治疗效果优于传统换药方法,其机制与该药物能促进EGF、bFGF和VEGF表达、抑制HIF-1α表达有关.     

Mechanism of Ginseng-Rhodiola rosea in treatment of myocardial ischemia-reperfusion injury based on network pharmacology and molecular docking北大核心CSCD

Aim To investigate the sites and mechanisms of action of Ginseng-Rhodiola rosea in the treat ment of myocardial ischemia-reperfusion injury ( MI-RI) via using network pharmacology approach, molecu¬lar docking techniques and experimental studies. Methods The active ingredients and targets of Gin¬seng-Rhodiola rosea were screened through the TCMSP database and literature supplementation, and the GEN-EC ARDS ,DISGENET and DRUGBANK databases were searched to obtain the targets of MIRI. Functional pro¬tein interaction networks (PPIs) and the STRING database were used to screen out core targets. The DAVID database was also selected for gene ontology functional analysis ( GO) and KEGG signaling pathway enrich¬ment analysis. Lastly, the preliminary validation was performed with the help of molecular docking techniques and experimental studies. Results Forty-three active ingredients and 348 potential targets of Ginseng-Rhodiola were obtained, and targets such as IL-6 , TNF-α and VEGFA were found to be closely related to MIRI, mainly involving TNF, PDK-Akt, HIF-1 and other signaling pathways.The molecular docking results showed that soysterol, ginsenoside rh2 and rhodioloside had good binding effects and high matching with IL-6, TNF-α,Caspase-3,VEGFA,MAPK1 and other targets, among which the best binding was between Caspase-3 and ginsenoside rh2. The results of the experimental study further showed that Ginseng-Rhodiola rosea could improve myocardial tissue necrosis after myocardial ischemia-reperfusion , reduce myocardial cell edema and vascular congestion, and decrease the expression levels of TNF-α and IL-6 in MIRI rats. Conclusions Ginseng-Rhodiola may modulate multiple targets such as IL-6,TNF-α, Caspase-3, VEGFA and MAPK1 through dousterol, ginsenoside rh2 and rhodiol glycosides to inhibit inflammatory response and oxidative stress, reduce cardiomyocyte damage and exert therapeutic effects on MIRI. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Effect of ammonium pyrrolidine dithiocarbamate (PDTC) on NF-κB activation and CYP2E1 content of rats with immunological liver injury

Context: Ammonium pyrrolidine dithiocarbamate (PDTC) is a potent inhibitor of nuclear factor-κB (NF-κB). Recent studies have shown that NF-κB plays an essential role in the regulation of genes whose products are involved in the pathogenesis of immunological liver injury.

Objective: To study the function of NF-κB in immunological liver injury of rat model and its effect on CYP2E1 content and metabolic activity.

Materials and methods: The present study investigated the effect of passivating NF-κB activation on CYP2E1 using Bacillus calmette Guérin (BCG)-induced immunological liver injury in Sprague–Dawley rats measured in terms of enzyme levels. The degree of hepatic injury of rats was measured by using biochemical parameters, hepatic tissue pathological changes, and physiological parameters. Protein localization of liver NF-κB was detected by immunohistochemical assay. Western blot analysis was used to detect the protein expression of NF-κB, IκBα, iNOS, and CYP2E1. The content of CYP2E1 of homogenate in the rat liver was detected by ELISA assay and the enzyme kinetics of CYP2E1 probe drug chlorzoxazone was evaluated by high-performance liquid chromatography (HPLC) assay.

Results: The results showed that BCG-pretreatment (125?mg/kg) significantly (p?p?in vivo. Moreover, PDTC (ED₅₀: 76?mg/kg) dose dependently inhibited down-regulation of CYP2E1 (p?Conclusion: Passivation of NF-κB can inhibit the down-regulation of CYP2E1 and iNOS to induce in rat liver tissue with immunological liver injury; NF-κB may be involved in the CYP2E1 regulation through iNOS.     

Effects of squalene on expression of LDLR in HepG2 cells and its mechanism北大核心CSCD

Aim To investigate the effect of squalene on LDLR expression in HepG2 cells and its mechanism of down-regulated cholesterol. Methods The proliferation of HepG2 cells exposed to squalene at different concentrations was measured by MTT assay. The effect of squalene on the expression of LDLR in HepG2 cells was measured by flow cytometry and fluorescence mi-croscopy. The effect of different concentrations of squalene on the interaction between SCAP and Insig2, two key protein molecules of SREBP pathway, was assayed by FRET technology. Results MTT results showed that squalene had inhibitory effect on the proliferation of HepG2 cells in a dose-dependent manner. Flow cytometry and fluorescence microscopy results showed that squalene enhanced LDLR expression in HepG2 cells compared with the control group. The results of FRET technology revealed that compared with model control group, the YFP fluorescence value in Squalene group dramatically declined, and the YFP fluorescence value of each drug group decreased with the range of 5-25 |xmol L1 squalene concentration. Conclusions Squalene may promote the expression of LDLR in HepG2 cells through inhibiting the interaction between SCAP and Insig2 proteins in SREBP pathway, which may confirm that squalene is a potential novel drug for the down-regulation of cholesterol level. © 2018 Publication Centre of Anhui Medical University. All rights reserved.     

Effect of a novel phosphodiesterase type 5 inhibitor,CPD1, on carbon tetrachloride-induced liver fibrosis in mice北大核心CSCD

Aim To investigate the effects of CPD1, a novel phosphodiesterase 5 inhibitor, on liver pathological phenotype and hepatic stellate cells (HSCs) activation in hepatic fibrosis model mice caused by carbon tetrachloride ( CCl4). Methods Male C57BL/6 J mice were divided into four groups randomly ( control group, CCl4group, CCl4+ CPD1 group and CCl4+ tadalafil group) . Hepatic fibrosis model was construc¬ted by intraperitoneal injection of CCl4( twice a week) . Four weeks after CCl4injection, the mice were treated with CPD1 (2 mg kg-1• d-1) , or Tadalafil (10 mg • kg-1• d-1) by intragastric administration, respec¬tively, for four weeks. Hematoxylin-eosin staining and Sirius Red staining were used to observe the distribu¬tion of liver tissue structural lesions and fibrosis. Im-munohistochemical staining was used to detect the ex¬pression of a-smooth muscle actin ( a-SMA) and fi-bronectin. Results Compared with control group, the liver tissue structure was seriously damaged in CCl4group with many hepatocytes necrosis and inflammatory cell infiltration, indicating that liver injury occurred in the CCl4-induced hepatic fibrosis model mice. Moreo¬ver, the expressions of a-SMA increased significantly in CCl4group. Compared to CCl4group, the liver tissue damage was significantly improved in PDE5 inhibitors group,most notably, CPD1 had a better curative effect than tadalafil did. Furthermore, CPD1 inhibited the ex¬pression of a-SMA markedly and reduced the expres-sion of ECM-related proteins induced by transforming growth factor pi ( TGF-f31 ) in Lieming Xu-2 ( LX-2 ) cells. Conclusions Phosphodiesterase 5 inhibitor CPD1 strongly alleviates CCl4-induced hepatic damage by inhibiting the activation of HSCs and expression of collagen fibers. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Effect of RAGE and its ligands on CD4+ T cells北大核心CSCD

RAGE (receptor for advanced glycation end products) is a multiligand receptor on the cell surface. Ligand-RAGE interactions activate several signal transduction pathways that propagate cellular oxidative stress and inflammatory response. RAGE expressed on the CD4+ T cells has been identified as a central transduction receptor which affects the activation, proliferation, migration and differentiation of the cells. In addition, blockade of RAGE suppressed the development of multiple immune-related disorders mediated by CD4+ T cells. These studies highlight the importance of RAGE and its ligands for CD4+ T cells. This article briefly reviews the role of RAGE and its ligands on the proliferation, migration and differentiation of CD4+ T cells and summarizes the related research progress.