1.Study on clinical relationships between gene polymorphism of vitamin D receptor and adiponectin with susceptibility of non-alcohol fatty liver disease
Zhoutao HE ; Canxia XU ; Xiangyang HAN ; Yiyao CHEN ; Yi CHEN
Chongqing Medicine 2017;46(12):1606-1609
Objective To discuss and analyze the clinical relationships between the gene polymorphism of vitamin D receptor and adiponectin with the susceptibility of non-alcohol fatty liver disease(NAFLD).Methods One hundred and two cases of NAFLD were selected as the observation group,and other 100 healthy volunteers were selected as the control group.The gene polymorphism of vitamin D receptor and adiponectin in the two groups was detected,then the genotype distribution and allele frequencies of vitamin D receptor and adiponectin were compared between the two groups,then their relationship with the susceptibility of NAFLD was analyzed.Results The genotype distribution situation of vitamin D receptor BsmI site,adiponectin 45 and 276 sites had statistically significant difference between the observation group and control group(P<0.05).The B allele frequency of BsmI site of vitamin D receptor in the observation group was far lower than that in the control group,and the allele frequencies of 45-T and 276-G of vitamin D receptor in the former were far higher than those in the latter,and the differences between the two groups were statistically significant(P<0.05).The multivariate unconditional Logistic regression analysis showed that vitamin D receptor:bb genotype,adiponectin 45 locus:TT genotype and ALT,TG,complicating hypertension history and HOMA-IR all were the independent risk factors in NAFLD patients.Conclusion The genotype distribution of different sites of vitamin D receptor and adiponectin has obvious abnormality in the patients with NAFLD,and both are closely related with the NAFLD susceptibility.
2.Expression and significance of ecto-5'-nucleotidase in colon tissue of Crohn's disease mice
SU Peiqiang ; ZHONG Zhuangxia ; CHEN Yiyao ; CHEN Chaochao ; HE Zhoutao ; LAN Cheng
China Tropical Medicine 2023;23(10):1071-
Abstract: Objective This study aimed to investigate the expression of CD73 in colonic tissues in Crohn's disease (CD) and its significance and possible mechanism of action. Methods Thirty male BALB/c mice were randomly divided into normal control group, model group and intervention group. The control group was fed normally, and the model group was treated with TNBS+40% alcohol enema to establish a mouse model of Crohn's disease induced by chronic inflammation. The intervention group was treated with AB-680 intraperitoneally on the second day of each enema based on the model group. Mice body weight, fecal traits and fecal occult blood were recorded for disease activity index (DAI) score of inflammatory bowel disease. The animals were sacrificed at 7th week, their colonic tissues were removed, weighed and measured. The tissue inflammation was observed by standard hematoxylin-eosin (HE) staining. Masson staining was used to measure the area of collagen in colon tissue of mice. CD73 was detected by fluorescence quantitative PCR and immunohistochemistry. The expression levels of TGF-β, TNF-α and IL-6 in colon tissue of mice were determined by ELISA. Results The DAI score was (0.10±0.16) in the control group, (2.80±0.79) in the model group, and (3.07±0.34) in the intervention group. Compared with the control group, the DAI scores of the model and intervention groups were increased significantly (P<0.05). Compared with the model group, the DAI score of the intervention group was significantly increased (P<0.05). HE staining showed that there was no inflammation in the colon of the control group, while the colon of the model group and the intervention group showed typical inflammatory manifestations such as edema and congestion, inflammatory cell infiltration, and mucosal ulcer. The area ratio of collagen in the control group was (4.95±0.82)%, in the model group was (24.62±1.46)%, and in the intervention group was (54.47±2.75)%. Compared with the control group, the area ratio of collagen in the model group and the intervention group was significantly increased (P<0.05). Compared with the model group, the area ratio of collagen in the intervention group was significantly increased (P<0.05). Compared with the control group, the expression of CD73 in colon tissue of the model group and the intervention group was significantly increased (P<0.05). Compared with the model group, the expression of CD73 in colon tissue of the intervention group was decreased (P<0.05). Compared with the control group, the expressions of TGF-β, TNF-α and IL-6 in the model group and the intervention group were significantly increased (P<0.05). Compared with the model group, the expression of TGF-β, TNF-α and IL-6 in the intervention group decreased (P<0.05). Conclusions CD73 is upregulated in colon tissue of CD mice, it can inhibit inflammatory reaction and improve fibrosis by up-regulating TGF-β expression. On the other hand, CD73 can aggravate the inflammatory response in CD intestinal inflammation and fibrosis by up-regulating the expression of IL-6 and TNF-α. Therefore, CD73 may play a bidirectional regulatory role in intestinal inflammation and fibrosis of CD.
3.Expression of obesity-related gene and forkhead transcription factors in liver of nonalcoholic fatty liver disease rat model
Yiyao CHEN ; Yi CHEN ; Zhoutao HE ; Manni CAI
Chongqing Medicine 2017;46(30):4185-4186,4189
Objective To research the obesity-related gene (FTO) and forkhead transcription factors O1 (FoxO1) protein expression level in the livers of non-alcoholic fatty liver disease(NAFLD) rat model. Methods The animal model of NAFLD in rats was prepared by feeding high energy and high fat feed. Then the rat blood and liver tissue were collected for detecting the liver index and blood biochemical indexes, including triglycerides (TG), total cholesterol (TC), alanine aminotransferase (AST), aspartate aminotransferase(ALT) ,alkaline phosphatase(ALP),high-density lipoprotein (HDL) and low density lipoprotein(LDL) ;the liver pathological examination was performed;FTO protein and Fox O1 protein expression levels in rat liver were detected by using the immunohistochemical assay. Results The rat liver weight,body weight and liver index after 8 weeks in the model group were higher than those in the control group,the difference was statistically significant (P<0.05);the levels of AST, ALT, LDL, ALP, TG and TC in the model group were higher than those in the control group,the difference was statistically significant (P<0. 05),the HDL level in the model group was lower than that in the control group, the difference was statistically significant (P<0.05) ;the model group produced steatosis and inflammation in hepatic lobule part,while the control group had no these lesions;the FTO prot ein and FoxO1 protein expression levels in liver of the model group were higher than those in the control group,the difference was statistically significant (P<0.05). Conclusion FTO and FoxO1 interaction may disturb the normal energy and fat metabolism.
4.Study on phenotype and function of intestinal gamma delta T lymphocytes in mice with post-infectious irritable bowel syndrome
Xuchun ZHOU ; Xiaoning SUN ; Bo YANG ; Baili HUANG ; Taozhi DENG ; Zhoutao HE ; Xiangyang HAN ; Sun DEMING ; Cheng LAN
Chinese Journal of Immunology 2016;32(7):957-961
Objective:To investigate the phenotype and function of the intestinal γδT lymphocytes in post-infectious irritable bowel syndrome mouse model. Methods:The mouse model for post-infectious irritable bowel syndrome was established by the infection with trichinella spiralis. The intestinal inflammation,abdominal withdrawal reflex( AWR) and colon transportation test were observed. 2 and 8 weeks later,the animals were sacrificed and the lymphocytes in the intestinal lymph nodes and spleen were collected,from which the γδT lymphocytes were isolated and purified by monoclonal antibody-immuno-microbeads method. The functions of the purified γδT lymphocytes were evaluated,including proliferation by 3 HTdR;CD69,CD62L molecule staining by flow cytometry. Furthermore,the con-centration of cytokine IL-17 and IFN-γ in the supernatant of the cultured γδT lymphocytes were detected by ELISA. Results: At 2nd weeks after infection,significant intestinal inflammation was observed,with increasingγδT lymphocytes,proliferating and activating with increasing production of IL-17. At 8th weeks after infection, the intestinal inflammation disappeared, whereas the number of γδT lymphocytes remained increasing,also with proliferating and activating with increasing production of IL-17. Meanwhile,the mice show higher AWR score and Bristol score. Conclusion: γδT lymphocytes could participate in the pathogenesis of PI-IBS via their proliferation,activation and production of IL-17.
5.Effect of GLP-1 on liver function and TLR4/NF-κB signal pathway in NAFLD rats
Yiyao CHEN ; Yi CHEN ; Zhoutao HE ; Manni CAI
The Journal of Practical Medicine 2018;34(1):26-29
Objective To study the effect of GLP-1 on the liver function and the signal pathway of TLR4/NF-κB in NAFLD rats.Methods Forty-five rats were randomly divided into two groups,15 with normal diet fed and 30 with high-fat diet,so as to form NAFLD model.In the tenth week,28 rats fed with high-fat diet were randomly divided into two groups,the model control group (14) and the GLP-1 intervention group (14 rats).The normal diet fed in 15 rats served as the normal control group.After the model group and the control group rats were injected with saline treatment,the intervention group rats were injected with liraglutide treatment.We observed the changes in the activity of rats,body weight,appetite,urine and other conditions.Results Compared with the control group,the liver index,liver mass,ALT,AST,TG and TC of the model rats were significantly higher than those of the control group (P < 0.05).The liver index,liver mass,ALT,AST,TG and TC of the model rats were significant lower in the GLP-1 intervention group than those in the model control group (P < 0.05).Compared with the normal control group,the expression of TLR4 and NF-κB protein in the model control group increased in the model control group (P < 0.05).Compared with the model control group,the expression of TLR4 and NF-κB protein was decreased in the GLP-1 intervention group (P < 0.05).Conclusion GLP-1 can significantly reverse the disorder of glucose and lipid metabolism and impaired liver function in NAFLD rats,and GLP-1 can significantly reduce the expression of NF-κB and TLR4 protein in the liver tissues of rats.
6.Expression of IFN-γ and IL-10 in peripheral blood and intestinal mucosa of patients with diarrhea post-infectious irritable bowel syndrome
Zhoutao HE ; Xiaoning SUN ; Xuchun ZHOU ; Baili HUANG ; Yiyao CHEN ; Taozhi DENG ; Xiangyang HAN ; Cheng LAN
Chongqing Medicine 2018;47(4):433-435
Objective To investigate the expression change of cytokines in peripheral blood and intestinal mucosa in the patients with diarrhea post-infectious irritable bowel syndrome(PI-IBS) and its relation with clinical symptoms scores.Methods Thirty outpatients and inpatients with diarrhea PI-IBS(observation group) and contemporaneous 30 individuals undergoing physical examination(control group) in the Hainan Provincial People's Hospital from January to December 2013 were selected.The peripheral blood mononuclear cells(PBMC) were separated and cultured.Then the levels of IFN-y and IL-10 in peripheral blood and cell culture supernatant fluid were detected by ELISA.The colonic mucosal tissue was taken by coloscopy.Then colonic mucosal IFN-γ and IL-10 protein expression was detected by immunohistochemistry staining.Furthermore,the correlationship between the level change of IFN-γ and IL-10 with clinical symptom score was analyzed by using the Spearman correlation method.Results Peripheral blod IL-10 and IFN-γ levels had no statistical difference between the two groups(P>0.05).Compared with the control group,in PBMC seperation and cuture,the IFN-γ level in the observation group was increased and IL-10 level was decreased,the difference was statistically signifieant(P<0.01).The intestinal main symptom score in the observation group had the positive correlation with IFN-γ expression level of PBMC culture supernatant fluid and colonic mucosal IFN-γ expression level(r=0.45,0.94,P<0.01),and had the negative correlation with IL-10 expression level(r=-0.52,-0.79,P<0.01).Conclusion The unbalance of IFN-γ and IL-10 level could be involved in the pathogenesis of diarrhea PI-IBS,which can serve as the observation indicators of disease activity.