甲泼尼龙联合环磷酰胺对博来霉素诱导的肺纤维化大鼠模型炎症反应与免疫细胞活性的影响CSCD

Objective To observe the effect of methylprednisolone (MP) combined with cyclo‑ phosphamide (CTX) on inflammation and immune cell activity in bleomycin (BLM)‑induced pulmonary fibrosis rat model. Methods Forty healthy 6 to 8‑week‑old SD rats were randomly divided into blank control, BLM model, BLM+MP, and BLM+MP+CTX groups, with 10 rats in each group. The rat model of pulmonary fibrosis was prepared by intratracheal infusion of BLM (5 mg/kg, only once). From the 7th day of modeling, MP (3 mg/kg) was injected in rats in the BLM+MP group and MP (3 mg/kg)+CTX (8 mg/kg) was injected via tail vein in rats in the BLM+MP+CTX group, once daily for 21 days. The degree of lung inflammation and fibrosis in rats was detected using HE and Masson staining methods. The numbers of granulocytes and neutrophils in bronchoalveolar lavage fluid (BALF) and blood T cell subsets in rats were detected using flow cytometry. Results On the 7th day of modeling, the external morphology, HE and Masson staining results of rat lung tissue showed that BLM‑induced pulmonary fibrosis model was successfully prepared. On the 28th day of modeling, the lung tissue structure of the BLM group was disordered with obvious collagen deposition, the number of granulocytes and neutrophils in BALF increased significantly, the propor‑ tion of blood T cells, CD4+ T cells, and regulatory T cells (Tregs) decreased, the proportion of CD8+ T cells, and the CD4+/CD8+ T cells ratio decreased significantly (all P<0.05). Compared with the BLM group, the degree of pulmonary fibrosis in the BLM+MP+CTX group was improved significantly, the number of granulo‑ cytes and neutrophils in BALF decreased significantly, the proportion of blood T cells, CD4+ T cells and Tregs cells increased significantly, the proportion of CD8+ T cells decreased, and the ratio of CD4+/CD8+ T cells increased significantly (all P<0.05). The improvement effect in rats of BLM+MP+CTX group was better than that of BLM+MP group, and the difference was statistically significant (P<0.05). Conclusion MP com‑ bined with CTX can reduce the degree of inflammatory reaction in rats with pulmonary fibrosis and improve T cell immune activity. © 2023 Chinese Medical Association. All rights reserved.     

Scopolin isolated from Erycibe obtusifolia Benth stems suppresses adjuvant-induced rat arthritis by inhibiting inflammation and angiogenesis

Objective To study the effects and mechanisms of scopolin isolated from the stems of Erycibe obtusifolia Benth in arthritis-associated inflammation and angiogenesis.Methods Adjuvant-induced arthritic rat,an animal model for human RA was used in this study for examining the potential remedial effect of scopolin.The swelling in both inoculated and non-inoculated paws,body weights and articular index(AI)scores were detected to evaluate the severity of the arthritis.Histologic assessment of tissue sections from rat ankles was also performed.Furthermore,the blood vessel density in the synovial tissues was quantitatively evaluated.In addition,expressions of VEGF,FGF-2,TNF-α,IL-1β and IL-6 in rat synovial tissues were determined by immunohistochemistry assay in an attempt to explain the mechanisms of scopolin for suppressing arthritis.Results Scopolin dose-dependently inhibited both inoculated and non-inoculated paw swelling in rat AIA.The mean AI scores of scopolin treated groups were also dose-dependently lower than that of model group.In addition,compared with the weights of model group,the mean body weights of rats treated with scopolin(50,100 mg·kg-1)were higher from day 13 to 22,perhaps indicative of healthier animals.The histologic architecture of the joint was highly abnormal in the model group rats,while high dose of scopolin treated rats preserved a nearly normal histologic architecture of the joint.Moreover,the new blood vessels were reduced dose-dependently in the synovial tissue of rat AIA treated with scopolin.Further,scopolin reduced the overexpression of IL-6,VEGF and FGF-2 in rat synovial tissues.Conclusions Scopolin is capable of reducing clinical symptoms of rat AIA by inhibiting inflammation and angiogenesis,and this compound may be a potent therapeutic agent for angiogenesis related diseases and can serve as structural base for screening for more potent synthetic analogs.     

corilagin protects non-alcoholic fatty liver disease in mice induced by high fat and high sugar diet via regulating AMPK-autophagy signaling北大核心CSCD

Aim To explore the effects of corilagin on non-alcoholic fatty liver disease induced by high-fat and high-sugar diet in mice via regulating AMPK-autophagy signaling. Methods Healthy 8-week-old male C57BL/6J mice were randomly divided into control group, model group and corilagin group. The mice of model group and corilagin group were fed with a high-fat and high-sugar diet for four weeks at the age of eight weeks. The corilagin group mice were also intraperitoneally injected with corilagin (20 mg • k g - 1 ), which was given once every 2 days for 4 weeks. The mice of the control group and the model group were given equal dose of normal saline. After modeling and administration, the mice were sacrificed and the liver weight recorded. The liver pathological changes of each group mice were assessed by HE staining, oil red O staining and Masson staining. The biochemical indexes in serum and liver tissue were detected by the ELISA kit. The p-AMPK and autophagy levels were detected by Western blot. Results The results showed that compared to control group, the liver weight of the model group increased, the AST and ALT levels in serum also significantly increased, there were a large number of fat vacuoles and severe lipid deposition and mild collagen fibrosis in liver, while the liver weight and TG level in liver significantly decreased, and the liver pathological changes were significantly improved after treated with corilagin. Western blot results showed the levels of autophagy related proteins such as Atg7 and Atg5 significantly decreased in the model group, and the p-AMPK level also significantly decreased. When treated with corilagin, p-AMPK and the autophagy levels were up-regulated. Conclusion corilagin can protect non-alcoholic fatty liver disease in mice induced by high fat and high sugar diet. The mechanism may involve increasing p-AMPK level and enhancing autophagy level in liver. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

The dynamic changes of serum tumor necrosis factor-α and the clinical significance in acute lung injury rats caused by mechanical trauma

Objective To determine the dynamic changes and mechanism of tumor necrosis factor-α(TNF-α) in serum in acute lung injury(ALI) induced by mechanical trauma in mechanical trauma rat model. Methods Totally 40 male Wistar rats were randomly divided into sham group and trauma group, sham-acute lung injury group and acute lung injury group. Noble-Collip drum was used to establish mechanical trauma rat model. Intraperitoneal injection of TNF-α established acute lung injury model. All rats after modeling of abdominal aortic blood sampling time points were killed and the serum levels of TNF-α were asasyed by ELISA. Results Serum TNF-α levels (334. 78 ±± 28) ng/L in the trauma group were significantly higher than those in the sham group( 177 ±10 ) ng/L (P ＜0. 01 ). The pathological results showed that both in trauma group and acute lung injury group lung infection were very obvious, while it was basically normal in both sham group and sham-acute lung injury group. Conclusions Serum TNF-αt levels of mechanical trauma may be associated with acute lung injury after mechanical trauma. Therefore, dynamic observation of the change of serum TNF-α after mechanical trauma will help the assessment and prognosis of the disease, it has important clinical significance.     

Ganodermalucidum polysaccharide peptides attenuate post myocardial infarction fibrosis through down-regulating TGF-β_1/SMAD signaling and ROS in mouse heart

OBJECTIVE Adverse cardiac fibrosis is an independent severe risk factor for heart failure and is the leading cause of death in myocardial infarction(MI)patients. The aim of this study was to investigate the potential effect of Ganodermalucidum polysaccharide peptides(GLPPs) on cardiac fibrosis after MI and the underlying mechanisms. METHODS C57 BL/6 mice MI model was set up by ligation of left anterior descending coronary artery. The MI and sham mice were orally gavaged with GLPPs(150 mg·kg~(-1)·d~(-1)). After 4 weeks of operation, cardiac function was detected by echocardiography. HE, Masson and Sirius Red staining was performed for histological damage and fibrosis assessment. Expression of FN, α-SMA, COⅢ, TGF-β_1, SMAD2/3, pSMAD3 were measured in mouse hearts by Western blotting.Primary rat neonatal cardiac fibroblasts(CFs) were stimulated with TGF-β_1(10 μg· L~(-1)) to establish fibrotic fibroblast model and were co-incubated with or without GLPP(6.25, 25 and 100 mg · L~(-1)) simultaneously for 48 h.Expression of FN, COⅢ, α-SMA, NOX4, SMAD2/3 and pSMAD3 were evaluated by Western blotting. Immunofluorescence staining of Ki67 assessed the cellular proliferation and phalloidin was used to label F-actin of the cytoskeleton to evaluate morphological change of CFs. H_2O_2(50 μmol · L~(-1), 24 h) was used to stimulate endogenous ROS production in CFs. GLPPs were co-incubated(100 mg·L~(-1)) simultaneously. Intracellular ROS level was detected with fluorescent probe DCFH-DA. RESULTS Echocardiography performed at 28 d after MI showed signs of cardiac remodeling, particularly a thickening of the interventricular septum and decreased ejection fraction compared with sham group. GLPPs group got a higher fractional shortening and ejection fraction than the MI group. Based on HE staining, the myocardium of GLPPs group exhibited less structural alterations. Masson and Sirius Red staining quantified the fibrosis and GLPPs alleviated cardiac fibrosis. Western blotting confirmed that GLPP reduced the expression of FN, COⅢ, α-SMA,TGF-β_1 and p SMAD3, suggesting that GLPPs lessened ECM deposition accompanied with the down-regulation of TGF-β_1/SMAD3 signaling. The fibrotic fibroblast markers and p SMAD3 were also suppressed by GLPPs dosedependently in vitro. Furthermore, GLPPs inhibited myofibroblast transdifferentiation induced by TGF-β_1, including the reduced cellular proliferation, the limited CFs migration and the maintenance of CFs morphology. GLPPs repressed NOX4 protein level in TGF-β_1 model and the fibrotic fibroblast markers induced by exposure of CF to H_2O_2 stimulation. Endogenous ROS production was elevated by H_2O_2 stimulation and were retarded by GLPPs. CONCLUSION GLPPs improved cardiac function and attenuated cardiac fibrosis post-MI, accompanied with less ECM deposition and down-regulated TGF-β_1/SMAD3 signaling both in vivo and in vitro. GLPP inhibited myofibroblast differentiation in TGF-β_1-treated CFs through modulation of ROS production produced by NOX4.     

The dynamic changes of serum tumor necrosis factor-α and the clinical significance in acute lung injury rats caused by mechanical trauma

Objective To determine the dynamic changes and mechanism of tumor necrosis factor-α(TNF-α) in serum in acute lung injury(ALI) induced by mechanical trauma in mechanical trauma rat model. Methods Totally 40 male Wistar rats were randomly divided into sham group and trauma group, sham-acute lung injury group and acute lung injury group. Noble-Collip drum was used to establish mechanical trauma rat model. Intraperitoneal injection of TNF-α established acute lung injury model. All rats after modeling of abdominal aortic blood sampling time points were killed and the serum levels of TNF-α were asasyed by ELISA. Results Serum TNF-α levels (334. 78 ±± 28) ng/L in the trauma group were significantly higher than those in the sham group( 177 ±10 ) ng/L (P ＜0. 01 ). The pathological results showed that both in trauma group and acute lung injury group lung infection were very obvious, while it was basically normal in both sham group and sham-acute lung injury group. Conclusions Serum TNF-αt levels of mechanical trauma may be associated with acute lung injury after mechanical trauma. Therefore, dynamic observation of the change of serum TNF-α after mechanical trauma will help the assessment and prognosis of the disease, it has important clinical significance.     

Triptolide inhibits NF-kappaB activation and reduces injury of donor lung induced by ischemia/reperfusion

Aim： To investigate the protective effect of triptolide （TRI） on ischemia/reperfusion- induced injury of transplanted rabbit lungs and to investigate the mechanisms underlying the actions of TRI. Methods： We established the rabbit lung trans- plantation model and studied lung injury induced by ischemia/reperfusion and the inhibitory effect of TRI on NF-KB. The severity of lung injury was determined by a gradual decline in PvO2, the degree of lung edema, the increase in the myeloperoxidase （MPO） activity, and the ultrastructural changes of transplanted lungs. The activation of NF-KB was measured by immunohistochemistry. The increase in intercellular adhesion molecule-1 （ICAM-1）, which is the target gene of NF-KB, was evaluated by ELISA. Results： After reperfusion, there was a gradual decline in the PvO2 level in the control group （group Ⅰ）. The level of PvO2 in the group treated with lipopolysaccharide （group Ⅱ） was significantly decreased, whereas that of the group treated with TRI （group Ⅲ） was markedly improved （P〈0.01）. In group III, the activity of MPO was downregulated, and the pulmonary edema did not become severe and the ultrastructure of the donor lung remained normal. The activity of NF-KB and the expression of ICAM- 1 was significantly increased in the donor lungs. TRI blocked NF-KB activation and ICAM-1 expression. Conclusion： The effects of TRI on reducing injury to donor lungs induced by ischemia/reperfusion may possibly be mediated by inhibiting the activity of NF-KB and the expression of the NF-KB target gene ICAM-1. Thus, TRI could be used in lung transplantations for improving the function of donor lungs.     

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.     

Pharmacokinetic-pharmacodynamic modeling of the anticancer effect of erlotinib in a human non-small cell lung cancer xenograft mouse model

Aim： Erlotinib is used to treat non-small-cell lung cancer （NSCLC）, which targets epidermal growth factor receptor （EGFR） tyrosine kinase. The aim of this study was to investigate the relationship between erlotinib plasma concentrations and phosphorylated EGFR （pEGFR） levels, as well as the relationship between pEGFR levels and tumor growth inhibition in a human non-small-cell lung cancer xenograft mouse model. Methods： Female BALB/c nude mice were implanted with the human NSCLC cell line SPC-A-1. The animals were given via gavage a single dose of erlotinib （4, 12.5, or 50 mg/kg）. Pharmacokinetics of erlotinib was determined using LC-MS/MS. Tumor volume and pEGFR levels in tumor tissues were measured at different time points after erlotinib administration, The levels of pEGFR in tumor tissues was detected using Western blotting and ELISA assays. Results： The pharmacokinetics of erlotinib was described by a two-compartment model with first order extravascular absorption kinetics. There was a time delay of approximately 2 h between erlotinib plasma concentrations and pEGFR degradation. The time course of pEGFR degradation was reasonably fit by the indirect response model with a calculated IC~o value of 1.80 pg/mL. The relationship between pEGFR levels and tumor volume was characterized by the integrated model with a Kbio value of 0.507 cm3/week which described the impact of pEGFR degradation on tumor growth. Conclusion： The pharmacokinetic/pharmacodynamic properties of erlotinib in a human tumor xenograft model were described by the indirect response model and integrated model, which will be helpful in understanding the detailed processes of erlotinib activity and determining an appropriate dosing regimen in clinical studies.     

Effects of ethanolic extracts of Euonymus alatus on CCl4-induced hepatic fibrosis in mice based on JAK2/STAT3 signaling pathway and its mechanism北大核心CSCD

Aim To explore the mechanism of ethanolic extracts of euonymus alatus on CCl4-induced hepatic fibrosis in mice by regulating JAK2/STAT3 signaling pathway. Methods Sixty C57BL/6J mice were randomly divided into control group,model group,EAL,EAM),EAH,and Silybin(n=10). Except for the control group,mice in other groups were injected with 25% CCl4 of 1.6 mL·kg-1 to induce HF model. Moreover,the positive group was administered 12.6 mg·kg-1 Silybin by gavage once a day,and EAL,EAM,and EAH were administered 72,140 and 280 mg·kg-1 ethanolic extracts of euonymus alatus once by gavage once a day,and the intervention lasted for six weeks. After 6th week,mouse blood was collected,the body weight and liver weight were measured and liver mass index calculated,the liver appearance was observed,and ALT,AST,TNF-α,IL-6,IL-1β were detected in serum. The protein expression levels of collagen Ⅰ,α-SMA,and JAK2/STAT3 signaling pathway-related protein(JAK2,STAT3,p-JAK2,pSTAT3)in mouse liver tissues were detected. Results Compared with the model group,EAM and EAH significantly decreased liver mass index,and the ALT,AST,α-SMA,collagen I levels of serum. After treatment,the liver morphology and structure,cellular inflammatory infiltration,fiber changes and collagen deposition in euonymus alatus intervention group were dose-dependently better than those of the model group. Compared with the model group,the expression of TNF-α,IL-6 and IL-1β in EAM and EAH serum decreased significantly(P<0.05). Compared with the model group,the protein expression levels of JAK2,STAT3,p-JAK2,pSTAT3 in EAM and EAH mouse liver tissues decreased significantly(P<0.05). Conclusion Ethanolic extracts of euonymus alatus have an anti-CCl4-induced HF effect in mice,and its mechanism may be related to the regulation of the JAK2/STAT3 signaling pathway. © 2023 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.     

The antitumor effect of cisplatin chemotherapy promoted by Taohong Siwu Decoction on mice with lung adenocarcinoma北大核心CSCD

Aim To study the antitumor effect of cispl-atin ( DDP) chemotherapy promoted by Taohong Siwu Decoction (TSD) on mice with lung adenocarcinoma mice. Methods Lewis lung carcinoma cell line was used to make homologous lung adenocarcinoma trans¬plantation mouse model. Normal control, Model, TSD, DDP, TSD + DDP groups were set up. The change of transplanted tumor volume after administration was observed, the weight of transplanted tumor was weighed, the expression of Ki67 in transplanted tumor tissue was detected by immunohistochemistry, TUNEL was detected by fluorescence staining, Bcl-2, Bax, cleaved Caspase-3 and cleaved Caspase-9 were detected by immunoblotting, and the content of D-dirtier in plasma was measured by ELISA. Results DDP plus TSD significantly inhibited the growth of transplanted tumor. Ki67 expression in tumor tissue was lower than that in DDP group (28. 3% ±3. 1% vs 40. 3% ±2.1% ). The combined use of TSD and DDP significantly promoted the apoptosis level of transplanted tumor. The positive rate of TUNEL was significantly higher than that of DDP group (41. 0% ±3.0% vs 30.7% ± 4.5%). Bax, cleaved Caspase-3 and cleaved Caspase-9 expressions in tumor tissue were also higher than those of DDP group, while the expression of Bcl-2 was significantly lower than that of DDP group. Moreover, we found a significant interaction between TSD and DDP on the expression of four apoptotic proteins ( P < 0.05 ) . The plasma D-dimer content in TSD + DDP group was significantly lower than that in DDP group (188. 50 ± 28. 46 vs 269.80 ± 35.92) μg • L-1(P < 0.05). Conclusion TSD may promote the inhibitory effect of DDP chemotherapy on transplanted lung adenocarcinoma by alleviating carcinoma hy-percoagulale state. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

The therapeutic effect of Balanophora polysaccharide on acetic acid gastric ulcer in rats and its mechanism; [蛇 ? 多糖对大鼠乙酸型胃溃疡的治疗作用 ? 机制]北大核心CSCD

Aim To study the therapeutic effect of Balanophora polysaccharide(BPS)on gastric ulcer(GU)induced by acetic acid in rats and to investigateits mechanisms. Methods Sixty male SD rats were randomly divided into sham-operated group, GU model group, omeprazole positive group(3.6 mg·kg-1), and low, medium and high dose of BPS treatment groups(100, 200 and 400 mg·kg-1). The GU model group was prepared by acetic acid cautery method, and the morphology and pathological changes of ulcers were observed by visual observation combined with HE staining, and the ulcer area and inhibition rate were measured and calculated; superoxide dismutase(SOD)activity, malondialdehyde(MDA)content and glutathione peroxidase(GSH-PX)activity were measured by enzymatic assay; tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)content were detected by ELISA. The expression levels of epidermal growth factor(EGF)and epidermal growth factor receptor(EGFR)were measured by immunohistochemistry staining and Western blot. Results Compared with the sham-operated group, obvious ulcer damage was seen in the model group. Compared with the model group, the BPS-treated group showed a significant reduction in ulcer area, an increase in SOD and GSH-PX activity and EGF and EGFR expression levels, and a significant decrease in MDA, TNF-α and IL-6 content. Conclusions BPS has a therapeutic effect on GU in rats, and its mechanism may be related to the inhibition of oxidative stress, suppression of inflammatory stimuli and promotion of regenerative repair of gastric mucosa. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Mechanism of Buyang Huanwu Decoction alleviating inflammatory response after cerebral ischemia/reperfusion injury by activating SIRT1 in hippocampus of rats北大核心CSCD

Aim To explore the inhibitory effect of Buyang Huanwu Decoction on the inflammatory response in the hippocampus of brain tissues of CIRI rats by regulating SIRT1 and the underlying mechanism. Methods The middle cerebral artery embolization (MCAO) model was prepared in rats and divided into sham operation group (Sham), model group (MCAO/R), Buyang Huanwu Decoction group (BYHWT),and BYHWT + SIRT1 inhibitor group (BYHWT + EX527). Zea Longa was used to detect the neurological function score of rats in each group; TTC staining was used to determine the volume of cerebral infarction; HE staining was used to observe the pathological damage of the hippocampus; Western blot was used to detect the expression levels of SIRT1 and IL-6; immunohistochemistry was used to detect TNF-α, IL-1β expression level. Results Compared with the sham group,the neurological function score of the MCAO/R group increased (P < 0.05); the volume of cerebral infarction increased (P < 0.05); the nerve cells in hippocampus were severely damaged, arranged disorderly, and the nucleus was broken; Western blot showed that the expression of SIRT1 decreased, IL-6 expression increased (P <0.05); immunohistochemistry showed that TNF-α,IL-1β expression increased (P < 0.05). Compared with the MCAO/R group, the neurological function score of the BYHWT group decreased (P <0.05); the volume of cerebral infarction decreased (P < 0.05); the damage of nerve cells in hippocampus was reduced; Western blot showed that the expression of SIRT1 increased and IL-6 expression decreased (P < 0.05); immunohistochemistry showed that TNF-α, IL-1β expression decreased (P < 0.05). Compared with the BYHWT group, the neurological function score of the BYHWT + EX527 group increased (P < 0.05); the volume of cerebral infarction was raised (P <0.05); the damage of nerve cells in hippocampus was aggravated; Western blot showed that the expression of SIRT1 decreased and IL-6 expression increased (P < 0.05); immunohistochemistry showed that TNF-α, IL-1β expression increased (P < 0.05). Conclusions Preliminary discussion of Buyang Huanwu Decoction can activate SIRT1 in hippocampus of rat brain tissues to reduce the inflammatory response after CIRI and play a role in brain protection. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

The protective effect of hesperidin on cardiac and renal tissue damage in DOCA/Salt hypertensive rats北大核心CSCD

Aim To investigate the protective effect of hesperidin (HES) on cardiorenal damage induced by DOCA/Salt hypertension and the underlying mechanisms. Methods Eighteen male SD rats were randomly divided into normal group (Ctrl), model group (DOCA/Salt), and DOCA/Salt with hesperidin group (DOCA/Salt + HES). HES was administered for four weeks. Blood pressure, serum creatinine and blood urea nitrogen were measured. The pathological changes in heart and kidney were examined by HE, Masson and Sirius red staining. The expression of α-SMA, collagen I and TGF-β were detected by Western blot. The mRNA levels of Nlrp3, TNF-α, IL-1β, IL-6 and NOXs were measured using qRT-PCR. Results Compared with the model group, HES administration significantly attenuated the occurrence of DOCA/Salt hypertension, improved renal function indicators of hypertensive rats, reduced renal and cardiac fibrosis, deduced the expression of α-SMA, collagen I and TGF-β, inhibited the expression of Nlrp3, TNF-α, IL-1β and IL-6, and decreased the expression of NOXs in renal and cardiac tissues. Conclusions HES can delay the occurrence of hypertension and protect against hypertension-induced renal and cardiac tissue damage, which may be related to the reduction of inflammatory reaction and oxidative stress by HES. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Effect of a novel phosphodiesterase type 5 inhibitor,CPD1,on unilateral renal interstitial fibrosis caused by ureteric obstruction in mice北大核心CSCD

Aim To investigate the effects of CPD1,a novel phosphodiesterase 5 inhibitor,on renal pathological phenotype and fibrotic protein expression in renal fibrosis model mice. Methods Male C57BL/6 J mice were divided into three groups randomly(sham group,UUO group and UUO+CPD1 group). Unilateral ureteric obstruction model was constructed by surgery,and CPD1(5 mg·kg-1·d-1)was administered by intragastric administration two hours after the modeling for seven days. HE and Sirius Red staining were used to observe the distribution of tissue structural lesions and fibrosis. Immunohistochemical staining and Western blot were used to detect the expression of fibronectin(FN),α-SMA,collagen-I and kidney injury molecule-1(Kim-1). Results Compared with sham operation group,the renal tubules of mice were dilated and accompanied by a large amount of inflammatory infiltration. Moreover,the expressions of FN,α-SMA,collagen-I and Kim-1 proteins increased significantly(P<0.05)in UUO group. CPD1 treatment improved the kidney structure and decreased the expression of collagen fibers. Furthermore,CPD1 inhibited the expression of FN,α-SMA,collagen-I and Kim-1 markedly(P<0.05). Conclusions Phosphodiesterase 5 inhibitor CPD1 alleviates the progression of renal fibrosis induced by unilateral ureteral obstruction through down-regulating ECM deposition in the extracellular matrix and expression of Kim-1. The specific mechanism remains to be further studied. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

LncRNA DACH1 protects against pulmonary fibrosis by binding to SRSF1 to suppress CTNNB1 accumulationOA

Idiopathic pulmonary fibrosis(IPF) is a progressive disease with unknown etiology and limited therapeutic options.Activation of fibroblasts is a prominent feature of pulmonary fibrosis.Here we report that lncRNA DACH1(dachshund homolog 1) is downregulated in the lungs of IPF patients and in an experimental mouse model of lung fibrosis.LncDACH1 knockout mice develop spontaneous pulmonary fibrosis,whereas overexpression of LncDACH1 attenuated TGF-β1-induced aberrant activation,collagen deposition ...     

MS80, a novel sulfated oligosaccharide,inhibits pulmonary fibrosis by targeting TGF-β1 both in vitro and in vivo

Aim： The pro-fibrogenic cytokine transforming growth factor-beta 1 （TGF-β1） has attracted much attention for its potential role in the etiology of idiopathic pulmonary fibrosis （IPF）. Here, we demonstrate that MS80, a novel sulfated oligosaccharide extracted from seaweed, can bind TGF-β1. The aim of the present study was to determine whether MS80 is capable of combating TGF-β1-mediated pulmonary fibrotic events both in vitro and in vivo, and to investigate the possible underlying mechanisms.
Methods： Surface plasmon resonance was used to uncover the binding profiles between the compound and TGF-β. MTI- assay, flow cytometry, Western blot analysis, BCA protein assay and SDS-PAGE gelatin zymography were used to probe the antifibrotic mechanisms of MS80. The in vivo fibrotic efficacy was evaluated in a bleomycin instillation-induced rat model. Results： We report that MS80, a new kind of sulfated oligosaccharide extracted from seaweed, inhibits TGF-β1-induced pulmonary fibrosis in vitro and bleomycin-induced pulmonary fibrosis in vivo. Our results indicated that MS80 competitively inhibited heparin/ HS-TGF-β1 interaction through its high binding affinity for TGF-β1. Moreover, MS80 arrested TGF-β1-induced human embryo pulmonary fibroblast （HEPF） cell proliferation, collagen deposition and matrix metalloproteinase （MMP） activity. Intriguingly, MS80 deactivated both the ERK and p38 signaling pathways. MS80 was also a potent suppressor of bleomycin-induced rat pulmonary fibrosis in vivo, as evidenced by improved pathological settings and decreased lung collagen contents.
Conclusion： MS80 in particular, and perhaps oligosaccharide in general, offer better pharmacological profiles with appreciably few side effects and represent a promising class of drug candidates for IPF therapy.     

α-Hederin inhibits growth of lung cancer A549 cells in vitro and in vivo by decreasing c-Myc and HIF-1α dependent glycolysis

OBJECTIVE α-Hederin is an effective component of the traditional Chinese medicine Pulsatilla chinensis,which has been reported to exert many pharmacological activities. However, the effect of α-hederin on metabolism is still unclear. This study aimed to illuminate the role of α-hederin in glucose metabolism in lung cancer cells and investigate the molecular mechanism of α-hederin. METHODS CCK8 and colony formation assays were employed to assess the anti-proliferative effects induced by α-hederin. Glucose uptake, ATP generation, and reduced lactate production were measured using kits, and an A549 tumor xenograft mouse model of lung cancer was used to assess the in vivo antitumor effect of α-hederin(5, 10 mg·kg~(-1)). Glycolytic-related key enzymes were detected by Western blotting and immunohistochemical staining. RESULTS Cell proliferation was significantly inhibited by α-hederin in a dose-dependent manner and that α-hederin inhibited glucose uptake and ATP generation and reduced lactate production. Furthermore, α-hederin remarkably inhibited hexokinase 2(HK2), glucose transporters 1(GLUT1), pyruvate kinase M2(PKM2), lactate dehydrogenase A(LDHA), monocarboxylate transporter(MCT4), c-Myc, and hypoxia inducible factor-1α(HIF-1α) protein expression. Using inhibitors, we proved that α-hederin inhibits glycolysis by inhibiting glycolytic regulators. Moreover, a tumor xenograft mouse model of lung cancer further confirmed that α-hederin inhibits lung cancer growth via inhibiting glycolysisin vivo. CONCLUSION α-Hederin inhibits the growth of non-small cell lung cancer A549 cells by inhibiting glycolysis.The mechanism of glycolysis inhibition includes α-hederin inhibiting the expression of the glycolytic regulatory factors HIF-1α and c-Myc.