1.Effects of phenylethanol glycosides from Cistanche tubulosa on proliferation of rat HSC induced by PDGF-BB and its mechanism
Shuping YOU ; Jun ZHAO ; Long MA ; Shilei ZHANG ; Tao LIU
Chinese Pharmacological Bulletin 2016;32(9):1231-1235
Aim To investigate the effect of phenyle-thanol glycosides from Cistanche tubulosa(CPhGs) on the proliferation and activation of rrPDGF-BB induced HSC and their target points for resisting hepatic fibro-sis,to elucidate the molecular mechanism in molecular level, and provide basic data for the further develop-ment of new drugs. Methods HSCs were cultivated by CPhGs with different concentrations ( 0 , 3. 91 , 7. 81 , 15. 63 , 31. 25 , 62. 50 , 125. 00 , 250. 00 , and 500 mg ·L-1 ) and IC50 of CPhGs was determined. CPhGs with different concentrations ( 25 , 50 , 75 , 100 mg · L-1 ) were selected, and after the cells were stimulated with rrPDGF-BB, cell proliferation was determined by MTT. ERK1/2 ,α-SMA, c-fos, c-jun and Collagen I mRNA and Erk1/2 ,P-Erk1/2 and CollagenⅠprotein ex-pressions were assayed by RT-PCR and Western blot. Results CPhGs of ( 50 ~100 ) mg · L-1 concentra-tions groups could effectively inhibit rrPDGF-BB-medi-ated proliferation(P<0. 05) and CPhGs of(25~100) mg·L-1 concentrations groups had no significant cyto-toxicity( P >0. 05 ) . CPhGs of ( 25 ~100 ) mg · L-1 concentrations groups could inhibit ERK1/2 ,α-SMA,c-fos, c-jun and CollagenⅠmRNA levels, and also ob-viously inhibited Erk1/2 ,P-Erk1/2 and Collagen Ⅰ pro-tein expression on HSC. Conclusions CPhGs has the protective effect against hepatic fibrosis. The mecha-nism of this process may involve the interference with PDGF/ERK1/2 signaling pathway and inhibiting the activation and proliferation of HSC.
2.Effect and mechanism of Cistanche phenylethanoid glycosides on rats with immunological liver fibrosis
Shuping YOU ; Jun ZHAO ; Long MA ; Tudimat MUKARAM ; Shilei ZHANG ; Tao LIU
Chinese Journal of Pharmacology and Toxicology 2016;30(5):504-510
OBJECTIVE To study the anti-fibrotic effect of Cistanche phenylethanoid glycosides (CPhG) in bovine serum albumin (BSA)-induced liver fibrosis in rats and its possible mechanism METHODS Seventy-five SD rats were randomly divided into six groups:normal control(distilled water-treated),model(BSA-treated),positive drug〔BSA-treated+compound Biejiarangan tablets(BJRG) 0.6 g·kg-1〕,and BSA-treated+CPhG(0.125,0.25 and 0.5 g·kg-1)groups. There were thirteen rats in each BSA-treated+CPhG(0.125,0.25 and 0.5 g·kg-1)group and twelve rats in other groups. Subcutaneous injection and tail vein injection of BSA immunity were used to induce the rat liver fibrosis model. Meanwhile, different therapeutic drugs were ig adminstered to rats. After the experimental period,rats were fasted for 12 h prior to 10%chloral hydrate administration and immediately euthanized. The liver was weighed to calculate the liver index. Glutamic-pyruvic transaminase (GPT),glutamic-oxalactic transaminase (GOT),alkaline phosphatase(ALP),total protein(TP)and albumin(ALB)were evaluated by the Mind-Ray automatic biochemical analyzer. The density of hydroxyproline (HyP) in liver tissues was determined using a spectrophotometric method according to the kit′s instructions. Histopathological changes and expressions of typeⅠ and typeⅢcollagens in liver tissues were also determined by immunohisto?chemical staining. RESULTS Compared with the normal control group,collagen fibers of liver tissues in the model group extended their links and enveloped the entire lobule,causing lobular structural damage and the formation of pseudolobules. The liver index(P<0.05),GPT,GOT,ALP,TP and ALB serum levels(P<0.05),HyP content(P<0.01)were significantly increased,so was the expression of typeⅠcollagens and typeⅢcollagens(P<0.01)in the model group. Compared with model group,various doses (0.125,0.25 and 0.5 g · kg-1) of CPhG significantly reduced the BSA-induced elevation of the liver index;GPT,GOT,ALP,TP and ALB serum levels(P<0.05),and HyP content decreased(P<0.01);the morphology of the pathological tissue sections was close to that of the normal control group,and CPhG significantly reduced the expression of two types of collagens(P<0.01). CONCLUSION CPhG can significantly reduce the degree of BSA-induced liver fibrosis in rats. The mechanism may be associated with down-regulation of two types of collagens and suppression of the activation of hepatic stellate cells.
3.Increasing Expression of IL-3 7 in Peripheral Blood Mononuclear Cell of Patients with Primary Biliary Cirrhosis and Its Clinical Significance
Tengda LI ; Yan CHEN ; Shuping LONG ; Mingli GU ; Peng LIU ; Linhong WU ; Shunjiang DENG ; Weiwei ZHANG ; Cheng QIAN ; Anmei DENG
Journal of Modern Laboratory Medicine 2017;32(1):12-15
Objective To test the expression level of IL-37 in peripheral blood mononuclear cells (PBMCs)of patients with primary biliary cirrhosis (PBC)and further explore its clinical significance in the pathological process of PBC.Methods Pe-ripheral blood samples were collected from 42 patients diagnosed as PBC and 38 health individuals examined at the same time during June 2013 to August 2015 in Changhai Hospital.PBMCs were separated by sucrose density gradient centrifugation, qualified Real Time-Polymerase Chain Reaction (qRT-PCR)was used to measure IL-37 mRNA expression level in PBMCs. Enzyme-Linked Immuno Sorbent Assay (ELISA)was to measure the protein level of IL-37,IL-6,IL-17,TNF-α,TGF-β,IL-18 and IL-23 in plasma.Meanwhile,the pathological stages of PBC cases were recorded.Pearson correlation analysis was performed on IL-37 and IL-6,TNF-α,IL-17,TGF-β,IL-18 and IL-23.Spearman rank correlation analysis was on IL-37 and pathological stages of PBC.Results The mRNA and protein level of IL-37 in experimental and controlled group were 2.81 ±0.94 vs 1.09±0.56,356.14±169.36 pg/ml vs 86.68±48.23 pg/ml separately(t=9.811,9.462,P<0.000 1),with sta-tistical differences.The correlation analysis showed that IL-37 was positively related with IL-17,TNF-α,IL-6 and TGF-β(r=0.561 2,0.661 9,0.672 1,0.765 3,P<0.001),and disease stages (Ⅰ~Ⅳ)(rs=0.348 9,P<0.05).Conclusion IL-37 might involve in the pathological process of PBC,and it is significant for disease prediction and diagnosis.
4.Affection of CD2 6/DPP4 on CD4+T Cells and Relative Cytokines in Patients with Crytococcal Meningitis and Its Clinical Significance
Tengda LI ; Shuping LONG ; Guixia XU ; Yun LIU ; Weiwei ZHANG ; Cheng QIAN ; Yuanlan HUANG ; Qin QIN ; Sunxiao CHEN ; Anmei DENG
Journal of Modern Laboratory Medicine 2016;31(5):38-41
Objective To analyze the affection and clinical significance of CD26/DPP4 on CD4+T cells and its cytokines in patients withCrytococcalMeningitis.Methods Peripheral blood was collected from 36 patients diagnosed withCrytococcal Meningitis in Changhai Hospital and Changzheng Hospital,Shanghai from August,2011 to December,2015,meanwhile 36 health controls’was also acquired.Peripheral blood mononuclear cell (PBMC)was separated by density gradient centrifuga-tion,CD26+CD4+T and CD26-CD4+T cell groups were classified by Flow Cytometry,the expression level of cytokines was tested by reverse transcriptase-polymerase chain reaction (RT-PCR).The correlation between DPP4 activity,CD26+CD4+T (%)and APACHE II score,IL-17,TNF-α,IL-4,IFN-γwas measured by Pearson coefficient.Results CD26+CD4+T(%)between experimental and control groups was 13.35±3.83 vs 8.39±2.14 (t=6.78,P<0.000 1).DPP4 activity was 50.89±17.21 mU/ml vs 73.83±20.24 mU/ml (t=5.18,P<0.000 1),with statistically significant differences.In ex-perimental groups,CD26+CD4+T (%)was positively related with APACHE II score,IL-17,TNF-α(r=0.431,0.564, 0.688,P=0.003 8,0.001,0.004 6).DPP4 activity was negatively interrelated with APACHE II score,IL-17,TNF-α,IFN-γ(r=-0.544,-0.489,-0.678,-0.734;P<0.001).Conclusion CD26/DPP4 may be involved in the pathogenesis of Crytococcal Meningitis through regulation of Th subgroups,and it was the potential therapeutic target and the predicted marker of the disease.
5.Elevated Expression of CD69 +CD4+CD25-T Cells in peripheral Blood from Patients with Autoimmune Pancreatitis and Its Clinical Significance
Tengda LI ; Shuping LONG ; Yuanlan HUANG ; Yi SUN ; Jie GUO ; Yun LIU ; Weiwei ZHANG ; Mingli GU ; Anmei DENG
Journal of Modern Laboratory Medicine 2017;32(4):29-31
Objective To test the expression level of CD69+CD4+CD25-T cells in peripheral blood from patients with autoimmune pancreatitis,and further analyze its clinical significance.Methods Peripheral blood samples from 32 patients with AIP diagnosed in hematological department,Changhai Hospital and 32 health individuals examined at the same time were collected from September 2014 to December 2016,they were classified as experimental and control groups,separately.Peripheral blood mononuclear cells (PBMCs) was acquired by density gradient centrifugation,CD69+ CD4 + CD25-T cells in PBMCs were tested by flow cytometry,and the expression level of cytokines in plasm was by ELISA.The comparison of varies between the two groups was measured by two independent samples' t test.The relationship between the two measurement data was measured by pearson correlation coefficient.Results The expression levels of CD69 + CD4 + CD25-T in experimental and control groups were 10.36%±3.68% vs 3.99%±1.45% (t=9.110,P<0.0001).The expression level of TGF-β was 399.86±121.88 vs 143.87±56.22 pg/ml (t=10.79,P<0.000 1),both with statistical significance.The levels of CD69+CD4+CD25-T in experimental was positively correlated with TGF-β (r=0.653,P<0.001) and negatively with IL-4,IFN-γ,IL-2 (r=-0.442,-0.567,-0.351,P<0.05) and there was statistical significance.Conclusion CD69+CD4 +CD25-T cells might involve the immunopathology of AIP and could be the potential biomarker for clinical diagnosis and therapy.
6.Short-term prognostic factors for Guillain-Barré syndrome in children
Jiayu HONG ; Xiaoyi LI ; Hongyuan DAI ; Na WANG ; Can LUO ; Long ZHENG ; Daokai GONG ; Ruiling ZHOU ; Wenjing LUO ; Bo HU ; Shuping LIU ; Jiajia YAO ; Zuneng LU
Chinese Journal of Neuromedicine 2020;19(5):504-507
Objective:To explore the short-term prognostic factors for Guillain-Barré syndrome (GBS) in children.Methods:The clinical data of children with GBS from 24 hospitals in 10 provinces/municipalities/autonomous regions in southern China (south of Huaihe River) from January 1, 2013 to September 30, 2016 were retrospectively analyzed. The factors affecting the short-term prognoses of children were explored.Results:In these 78 children (50 males and 28 females), the average age was 9.53±5.44 years, and 19 were under 5 years old. Fifty children had history of prodromal events; 28 children had cranial nerve involvement, and 22 had autonomic nerve involvement. Five children needed assisted respiration, and one died during hospitalization. There was no statistically significant difference in percentage of children having poor short-term prognosis (scores of Hughes GBS disability scale≥3 at discharge) between children with different ages, children having different days from onset to admission, children with different clinical classifications or electrophysiological classifications, children with different treatment plans, children having presence or absence of prodromal events, children having presence or absence of cranial nerve involvement ( P>0.05). The proportion of children having poor short-term prognosis in children with autonomic nerve involvement was significantly higher than that of children without autonomic nerve involvement (31.8% vs. 10.7%, P<0.05). Conclusion:The short-term prognosis of children with autonomic nerve involvement is poor.