2.Study on the mechanism of action of total flavonoids of Litsea coreana for reducing blood glucose level in rat with type 2 diabetes mellitus.
Yu-Xiu SUN ; Yun-Xia LU ; Ling-Yun WANG
Chinese Journal of Integrated Traditional and Western Medicine 2010;30(6):617-621
OBJECTIVETo explore the possible mechanism of total flavonoids of Litsea coreana (TFLC) on reducing blood glucose level in rat with type 2 diabetes mellitus (T2DM).
METHODSMale SD rats of T2DM allocated in two groups were treated with 400 mg/kg TFLC or metformin respectively via gastrogavage for 6 weeks. Blood routine biochemical indices in rats were measured; pathology of rats' liver was examined with HE stain under transmission electron microscopy; levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in liver homogenate were determined, and the expression of protein tyrosine phosphatase 1B (PTP1B) in liver was detected using RT-PCR at the terminal of the experiment.
RESULTSBiochemical measuring showed that the glucose tolerance of rats after treatment was markedly improved in both groups. Meantime, levels of fast blood glucose (FBG), glycohemoglobin (HbA1c), fast blood insulin (FINS), free fatty acid (FFA), total cholesterol (TC), triglyceride (TG) and low density lipoprotein-cholesterol (LDL-C), as well as MDA level in liver were decreased, while levels of high density lipoprotein-cholesterol (HDL-C) in blood and SOD in liver were significantly increased in both groups after treatment, showing insignificant difference between two treatment groups. Light microscopic examination showed markedly fatty degeneration of liver, and electron microscopic examination found mitochondria swelling and endoplasmic reticulum breakage in liver of T2DM rats, but these changes were ameliorated to some extent after treatment. The elevated PTP1B expression in liver of T2DM rats was decreased in the TFLC treated group, but unchanged in the metformin treated group.
CONCLUSIONTFLC can significantly decrease the blood levels of glucose and lipid and ameliorate oxidation stress in liver; its mechanism of action in improving insulin resistance might be related with its suppression on PTP1B expression in rat's liver to enhance the insulin signaling pathway.
Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetes Mellitus, Type 2 ; drug therapy ; Flavonoids ; isolation & purification ; therapeutic use ; Hypoglycemic Agents ; therapeutic use ; Litsea ; chemistry ; Liver ; metabolism ; Male ; Oxidative Stress ; drug effects ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; metabolism ; Rats ; Rats, Sprague-Dawley
3.Effects of iodine excess on TPO and NIS genes mRNA expression in rats
Jian-ying, XIA ; Jing-jing, SUN ; Xiu-mei, ZHANG ; Dan, YUE ; Cui-yao, WANG
Chinese Journal of Endemiology 2008;27(2):145-148
Objective To observe the effects of iodine excess on thyroid morphology,the expression of thyroid peroxidase and sodium iodide symporter mRNA and to explore their mechanisms.Methods One-month SD rats were divided into three groups:control iodine(CI),high iodine Ⅰ(HI Ⅰ)and high iodineⅡ(HI Ⅱ)and were fed with water containing iodine in different concentrations by adding K103(5,5000,10 000μg/L)respectively.Rats were sacrificed after being fed for six months.The morphology of thyroid was investigated under light microscopy and electron microscopy,the serum thyroid hormones and ratio of TPO/β-actin and NIS/β-actin were measured by radio-immunoassay and RT-PCR method.Results The major changes were increased follicles with colloid accumulation in HI groups.The levels of serum thyroid hormones TT3 and TT4 were decreased gradually from CI[(75.68±13.99,1.45±0.49)nmol/L]to HI Ⅰ[(73.82±16.48,1.34±0.31)nmol/L]and HIⅡ groups[(70.65±11.43,1.15±0.39)nmol/L],but there were no significant differences among three groups(F=O.371,l.163,P>0.05).The TPO and NIS mRNA expressions in HI Ⅰ(1.28±0.10,0.56±O.17)and HI Ⅱ(1.14±0.04,0.39±0.06)were significantly lower(F=30.863,62.62.675,P<O.05)than those of control group(1.39±0.08,0.71±0.13).Conclusions Chronic iodine excess leads to definite histological changes in rat thyroid,and inhibits the expressions of TPO and NIS mRNA as well as thyroid hormone synthesis,which in turn acts as a protective mechanism against iodine excess.
4.Expression of GPX1-mRNA and apoptosis related signal molecular in Keshan disease patients
Rui-xia, SONG ; Yong-min, XIONG ; Xiu-zhen, ZOU ; Xiao-hong, DU ; Wen-yan, SUN
Chinese Journal of Endemiology 2010;29(4):359-361
Objective To investigate the meaning of expression of apoptosis related molecules NFKBp65 and p53 and GPX1-mRNA in patients with Keshan disease(KSD).Methods Sixteen chronic Keshan Disease patients were enrolled in KSD group according to electrocardiogram,chest X ray film and clinical examinations on 15,September in 2009,and 23 healthy people were included in control group from physical examination taken in The Second Affiliated Hospital of Xi'an Jiaotong University.Fresh blood(5 ml)was collected from antecubital vein of all subjects in the fasting state.Total mRNA and protein of blood sample were isolated using Trizol.GPX Assay Kit was used to detect GPX enzyme activity,and GPX1-mRNA expression was determined by SYBR Real-Time PCR.Meanwhile,expression of apoptosis related molecules NFKBp65 and p53 were determined by Western blot.Results GPX enzyme activity decreased significantly in KSD group[(108.61±14.10)U]compared with control group[(122.78±11.89)U,t=2.874,P<0.05],GPX1-mRNA level of KSD group(0.553±0.299)notably KSD group(0.802±0.057)compared with control group[(1.065±0.355),t=6.829,P<0.01].p53 increased in KSD group(1.604±0.191)compared with control group[(1.137±0.186),t=3.033,P<0.05].Conclusiom Decreased GPX1-mRNA expression may result in lower GPX enzyme activity of patients with KSD.Thus oxidative damage increases and cadioeyte apoptosis is activated by activating apoptosis signal pathway.
6.Bone marrow mesenchymal stem cells modulated the inflammatory response by regulating the expression of IL-4 and RAGE products in the rats with MODS
Xia ZHOU ; Guanghui XIU ; Yichao ZHU ; Xiaolei CHEN ; Wei XIONG ; Xinghua PAN ; Jie SUN ; Bin LING
Chinese Critical Care Medicine 2017;29(4):294-299
Objective To investigate the underlying mechanism of bone marrow mesenchymal stem cells (BMSC) modulating the inflammatory response during the multiple organ dysfunction syndrome (MODS), especially the expression of inflammatory cytokines, which will provide new theoretical and experimental basis of MODS in clinic. Methods BMSC of Sprague-Dawley (SD) rat (female, 4 weeks) was extracted and cultivated, and the 4th passage were used in experimental study. According to the random number table, 60 female SD rats were divided into three groups (n = 20 per group): sham group, MODS group, BMSC group. MODS model in rats was induced by lipopolysaccaride (LPS, 1 mg/kg) via femoral vein injection. Sham group was injected with the sterile phosphate buffer saline (PBS) in the same volume. BMSC group, in which BMSC infusion was started at 2 hours after 0.5 mL LPS stimulation (1×106/cells) through the tail vein. The survival rate was observed after 72 hours in each group. Abdominal aortic blood was collected for routine blood and biochemical examination at 72 hours after operation. Protein microarray was used to detect the related 34 inflammatory cytokines. Signal ratio was defined as the differentially expressed factors when it was more than 2.0 or less than 0.5. And enzyme linked immunosorbent assay (ELISA) was be applied to validate the significant inflammation factor. Meanwhile, the heart, kidney, intestine tissue was harvested, then their pathological changes were observed by hematoxylin eosin (HE) staining.Results 20, 12, 16 rats lived in sham group, MODS group and BMSC group respectively at 72 hours after operation. Compared with the sham group, the indicators (routine blood, liver and kidney function, myocardial enzyme) were apparently unusual, and the heart, kidney, intestine tissue were injured obviously in the MODS group. After BMSC administration, the organ function was improved and tissue damaged was alleviated significantly. Protein microarray showed that interleukin-4 (IL-4) and receptor for advanced glycation end products (RAGE) were significantly different in 34 goal cytokines. The signal ratio change of IL-4 was 0.397, 1.124, 2.826 respectively, and the signal ratio of RAGE was 6.197, 1.552, 0.250, respectively in MODS/sham group, BMSC/sham group, BMSC/MODS group. ELISA validated the result that the expression level of IL-4 decreased significantly (ng/L:3.59±1.21 vs. 29.10±5.78) and the expression level of RAGE increased significantly (ng/L: 1.09±0.04 vs. 0.11±0.03) in MODS group as compared with sham group (bothP < 0.05). Compared with the MODS group, the level of IL-4 was obviously higher than that in BMSC group (ng/L: 9.59±2.21 vs. 3.59±1.21,P < 0.01), and RAGE decreased significantly (ng/L: 0.29±0.07 vs. 1.09±0.04,P < 0.05).Conclusions BMSC administration can regulate the expression of IL-4 and RAGE in the rats subjected to MODS. Moreover, BMSC can promote the restoration of tissue and organ function, thus improve the survival rate. BMSC may be the target in cell therapy for the inflammatory disease.
7.Improvement of cerebral blood perfusion by L-argi nine following subarachnoid hemorrhage in rats
Baoliang SUN ; Suming ZHANG ; Xincheng WANG ; Zuoli XIA ; Hui YUAN ; Jian ZHANG ; Ruijua XIU
Chinese Pharmacological Bulletin 2003;0(12):-
AIM To determine the effect of L-arginin e on cerebral blood perfusion following subarachnoid hemorrhage(SAH) in rats. METHODS Endovascular perforating SAH models were replicated in Wista r rats, and animals were divided into sham-operated group, SAH group and SAH+ L-arginine group. Dynamic changes of regional cerebral blood flow (rCBF) with in 24 hours were measured and serum nitric oxide(NO, NO - 2/NO - 3)levels at different time points within 24 hours were detected. Mean arterial blood pre ssure and blood gas were monitored during the experiment. RESULTS No obvious change in physiological parameters in the three groups was observed . rCBF and serum nitric oxide level at every time point after operation in SAH g roup were lower than those in sham-operated group. Pathological alterations abo ve in SAH+L-arginine group were less obvious than those in SAH group. CONCLUSION L-arginine, by antagonizing the decrease of nitric oxi de, exerts protective effect on secondary cerebral ischemia following SAH.
8.Effects of pilose antler polypeptide on the glycosaminoglycan and type II collagen in experimental knee osteoarthritis.
Chun-Xia ZHANG ; Lei SUN ; Zhong-Biao XIU
China Journal of Orthopaedics and Traumatology 2012;25(2):138-142
OBJECTIVETo observe the influence of Pilose antler polypeptide on the glycosaminoglycan and type II collagen in the articular cartilage in experimental knee osteoarthritis.
METHODSTotally 64 New Zealand white rabbits of 6 months old were randomly divided into 2 groups:normal group (n = 8) and model group (n = 56). Model group was surgically induced into osteoarthritis model by method of Hulth. After successful modeling, the rabbits of model group were further divided into 2 groups: Pilose antler polypeptide-treatment group and control group, 24 rabbits in each group. Pilose antler polypeptide-treatment group received 0.5 ml intra-articular injection of Pilose antler polypeptide dilution liquid once in per 2 days for 30 days, while control group received 0.5 ml intra-articular injection of physiological saline. On days 7, 15 and 30 after intervention, articular cartilage samples were collected respectively. The content of glycosaminoglycan in articular cartilage was observed by toluidine blue staining and the expression of type II collagen in cartilage matrix was detected by immunohistochemical staining.
RESULTSAlong with the prolonging of time, the content of glycosaminoglycan and type II collagen in cartilage matrix of the Pilose antler polypeptide-treatment group and control group decreased gradually. On days 7, 15 and 30 after intervention, integrated optical density of the type II collagen positive area in cartilage matrix of the Pilose antler polypeptide-treatment group were (312.06 +/- 14.12), (273.31 +/- 12.42) and (248.34 +/- 10.41), which had statistically significant differences. Integrated optical density of the type II collagen positive area in cartilage matrix of the control group were (253.47 +/- 15.53), (215.67 +/- 9.72) and (160.01 +/- 13.23), which had statistically significant differences. At the same period, integrated optical density of the type II collagen positive area in cartilage matrix of the Pilose antler polypeptide-treatment group was higher than that of control group, which had statistically significant difference.
CONCLUSIONPilose antler polypeptide can inhibit reduction of the glycosaminoglycan and type II collagen in cartilage matrix and delay the degeneration of articular cartilage.
Animals ; Antlers ; chemistry ; metabolism ; Collagen Type II ; metabolism ; Disease Models, Animal ; Female ; Glycosaminoglycans ; metabolism ; Humans ; Male ; Osteoarthritis, Knee ; drug therapy ; metabolism ; Peptides ; metabolism ; pharmacology ; Rabbits
9.Changes of Renal Hemodynamic,Urine 6-Keto-Prostaglandin F_(1?),Thromboxan B_2 in Children with Henoch-Schonlein Purpura
shu-hua, WANG ; xiu-qiao, TIAN ; hui-sheng, SUN ; li-xia, LIANG ; zhan-ru, YIN ; yi, YANG
Journal of Applied Clinical Pediatrics 1992;0(05):-
Objective To investigate the characteristics of renal hemodynamic and the esoteric prostacyclin(PGI2),thromboxane A2(TXA2)level in children with early Henoch-Schonlein purpura(HSP),and study the function of TXB_2/6-Keto-prostaglandin F(6-Keto-PGF_(1?))(T/K)numerus in early changes of kidney injury.Methods Children involved in the experiment were dicided into 3 groups.Thirty-one patients with HSP,divided into 2 groups according to routine urianlysis:children with HSP without renal damage group(n=16)and Henoch-Schonlein purpura nephritis(HSPN)group(n=15).Control group with 16 healthy children,their age and sex match with the other 2 groups.The urine of all children,including the children in control group,was sampled in 24 hours.The urinary production of the samples were kept in the freezer at-20 ℃.The radioimmunoassay was applied to determine the 6-Keto-PGF_(1?),TXB_2 quantitatively,and calculate the number of T/K.In the early morning the children accept the Doppler arteria renalis sonography with an empty stomach to determine the Vmax of the period of contraction of the arteria renalis the Vmin of diastolic phase and the resistent index(RI).SPSS 13.0 software was used to analyze the data.Results 1.The renal hemodynamic indicated a change of high velocity and resistance,the masculine rate(83.9%)was ob-viously higher than that in routine urinalysis(48.4%)(?2=5.79 P0.05).The RI in the former group(0.798?0.165)was much higher than that in the other one(0.637?0.116)(t=4.02 P
10.Clinical features, gene mutation and genetic counseling in Rett syndrome patients
Xin-Li BAI ; Xiu-Xia WANG ; Hui-Feng ZHANG ; Yan-Xia SUN ; Zhen-Zhong LI
Chinese Journal of Applied Clinical Pediatrics 2013;28(8):592-595
Objective To investigate the mutations in methyl-CpG-binding protein 2 gene (MECP2 gene) from typical sporadic Rett syndrome patients,explore the correlations between their genotype and phenotype,assist in genetic counseling.Methods Genomic DNA was extracted from peripheral blood leukocytes from 2 patients and their parents using standard protocols.Polymerase chain reaction and direct sequencing were performed using specific primers from 4 exons in MECP2 gene.Results No mutations were found in exon 1,2,3.Two different heterozygous missense mutations in exon 4 within MECP2 gene were identified from 2 patients.Their nuclear acid changes were:c.C473T and c.C397T,leading to amino acid change accordingly:p.T158M and p.R133C.There were no same mutations from their parents.Phenotype of patient with c.C397T was milder than patient with c.C473T.Conclusions Most of typical Rett syndrome patients had mutations in MECP2 gene.Gene test should be performed.Their biological parents should be detected accordingly if the patient had positive found to support genetic counseling.