1.Effects of ω-3 polyunsaturated fatty acids intervention on Toll-like receptor 4 and high mobility group box 1 with intestinal ischemia/reperfusion injury
Guizhen HE ; Kaiguo ZHOU ; Xuefeng CHEN ; Yukang WANG ; Fie WANG
Chinese Journal of Clinical Nutrition 2013;(3):147-156
Objective To investigate the expressions of Toll-like receptor 4 (TLR4) and high mobility group box 1 (HMGB1) expression on distant tissue during the intestinal ischemia/reperfusion and the effects of ω-3 polyunsaturated fatty acids (ω-3 PUFAs) intervention in rats.Methods Forty-eight Sprague-Dawley male rats,weighing (281.50 ± 22.68) g,were randomly divided into three groups (n =16) after gastrostomy:normal diet (N) group,enteral nutrition (EN) group and EN plus ω-3 PUFAs (PUFA) group.Each group was further divided into lymph drainage (I/R + D) and non-drainage (I/R) sub-groups (n =8 each) according to whether treated with intestinal lymph drainage.All the rats were subjected to 60 min ischemia by clamping the superior mesenteric artery,followed by 120 min reperfusion,while the rats in the I/R + D subgroups were treated with intestinal lymph drainage for 180 min at the same time.Results The interleukin-6 level in lymph in N (I/R + D) group was significantly higher than in the EN (I/R + D) and PUFA (I/R + D) groups (PUFA vs EN vs N:(154.57 ±69.30) ng/L vs (97.58 ±40.34) ng/L vs (85.35 ±23.93) ng/L,P =0.021).Besides,the serum level of HMGB1 in PUFA (I/R + D) group was significantly lower compared to the other 5 groups [PUFA (I/R) vs EN (I/R) vs N (I/R) vs PUFA (I/R + D) vs EN (I/R + D) vs N (I/R + D):(2.95 ± 1.17) μg/L vs (3.86 ±0.99) μg/L vs (4.45 ± 1.73) μg/L vs (1.71 ±1.41) μg/Lvs (2.11±0.56) μg/Lvs (3.13 ±0.79) μg/L,P=0.000],and it also decreased in the PUFA (I/R) and EN (I/R) groups than the N (I/R) group (respectively,P < 0.05).Furthermore,the serum endotoxin level in PUFA (I/R) group was significantly lower compared to the N (I/R) and EN (I/ R) groups[PUFA(I/R) vsPUFA (I/R+D) vsEN (I/R) vs N (I/R):(0.020±0.004) EU/mlvs (0.028 ±0.006) EU/ml vs (0.028 ±0.005) EU/ml vs (0.018 ±0.006) EU/ml,P=0.014].Together the serum tumor necrosis factor-α level in both PUFA (I/R) and PUFA (I/R + D) groups were significantly lower than theEN (I/R),N (I/R) and N (I/R+D) groups [PUFA (I/R+D) vs PUFA (I/R) vs EN (I/R) vsN (I/R) vs N (I/R+D):(12.03 ±6.57) ng/L vs (14.32 ±6.11) ng/Lvs (23.27 ±15.60)ng/L vs (27.42 ± 10.37) ng/L vs (26.87 ± 5.30) ng/L,P =0.013].The jejunum and ileum mucosa in all the I/R groups showed swelling and atrophy and appeared fragile,while the PUFA groups showed less yellow staining and injury than the other two groups (P < 0.05,respectively).In addition,the expressions of TLR4 mRNA in jejunum,ileum,and liver in all the drainage groups were respectively lower than the corresponding non-drainage groups [jejunum:PUFA (I/R) vs EN (I/R) vs N (I/R) vs PUFA (I/R+D) vs EN (I/R+D) vsN (I/R+D):2.32±0.62vs3.08±1.29vs3.50±2.44vs 1.62±0.79vs 1.67±1.11 vs 1.94±0.81,P=0.025; ileum:PUFA (1/R) vsEN (1/R) vsN (1/R) vs PUFA (1/R+D) vsEN (1/R+D) vs N (1/R+D):2.67±1.08 vs 5.22 ± 3.96 vs 6.95 ±4.92 vs 1.70±0.68 vs 1.80±0.29 vs3.68±1.47,P=0.012; liver:PUFA (1/R)vsEN (1/R)vsN (1/R)vs PUFA (1/R+D)vsEN (1/R+D)vsN (1/R+D):5.67 ±1.94 vs 7.50 ±3.89 vs 7.18 ±4.55 vs 1.70 ±0.86 vs 3.90 ± 1.95 vs 4.12 ±2.11,P =0.001],which was consistent with the reduction of HMGB1 and the decrease of nuclear factor-κB activity in intestine,liver,and lung (P =0.000).Conclusions Lymph drainage and ω-3 PUFAs intervention can reduce the production of HMGB1 and inflammation factors,inhibit the expression of HMGB1 and TLR4 mRNA,and thus alleviate distant tissue injury caused by intestinal L/R.
2.The expression and clinical significance of Toll-like receptor 7 and 9 in peripheral blood B cells of patients with systemic lupus erythematosus
Tao WANG ; Zhijun LI ; Baiqing LI ; Linjie CHEN ; Fie TANG ; Changhao XIE ; Yongjun MEI ; Jing CHEN
Chinese Journal of Rheumatology 2009;13(8):560-562
Objective The aim of this study is to examine the expressions of Toll like receptor (TLR) 7 and TLR9 in the peripheral blood B lymphocytes of SLE patients and to analyze the correlation between TLR7/9 and clinical parameters. Methods lntracellular expression of TLR7/9 in the peripheral blood CD19+Blymphocytes was analyzed in 50 SLE patients and 30 healthy controls by flow cytometry. The difference of intracellular TLR7/9 expression levels in two groups was compared. Furthermore,the correlation between TLR7/9 expression and clinical parameters such as ESR, CRP, complement 3 (C3), complement 4 (CA), the level of serum IgG, anti-double stranded DNA antibody, anti-nuclear antibodies, SLEDAI score and urine protein excretion level, were analyzed. Results Compared with healthy subjects, the proportion of B cells expressing TLR7 and TLR9 was higher among SLE patients. Positive correlation was observed between TLR7 expression levels and clinical measurement of the SLEDAI and ESR. Negative correlation was observed between TLR7 expression levels and serum C3 levels. Positive correlation was observed between TLR9 expression levels and SLEDAI scores. Negative correlation was observed between TLR9 expression levels and serum C3 levels. Conclusion TLR7 and TLR9 expression is increased in the peripheral blood B cells of SLE patients, and correlates well with clinical parameters.
3.Expression and clinical implications of hMSH2 gene in sporadic insnlinomns
Mei MEI ; Yuanjia CHEN ; Chongmei LU ; Liming ZHU ; Haiyan WU ; Xin LU ; Xinting SANG ; Zhiying YANG ; Miao YU ; Hongding XIANG ; Fie CHEN
Chinese Journal of Pancreatology 2009;9(1):5-8
Objective To investigate the role of hMSH2 in the pathogenesis of sporadic insulinomas and to determine whether the expression of hMSH2 could be used to differentiate benign sporadic insulinomas from malignant ones. Methods Fifty-five sporadic insulinomas (40 benign and 15 malignant tumors) resected from 50 patients were obtained. Expression of hMSH2 was detected by immunohistochemistry staining. DNA was obtained from micradissected tissue. Loss of heterozygnsity (LOH) of hMSH2 gene was detected by PCR-LOH. 6 microsatellite markers were selected on 3 chromosomes, and microsatellite instability (MSI) status of tumor tissue were detected by PCR. The findings were analyzed in relation to the clinicopathological characteristics. Results Down-regulation of hMSH2 expression was found in 13% of 55 sporadic insulinomas. LOH of the hMSH2 gene was not present in 55 insulinomas. High frequency MSI (MSI-H, MSI occurred in at least 2 out of 6 sites) was present in 36% (20/55) of all the insulinomas. Down-regulation of hMSH2 expression was found in 33% of the 15 malignant tumors, while it was 5% in benign tumors (P < 0. 05). Conclusions Down-regulation of mismatch repair gene hMSH2 may be correlated with the degree of tumor malignancy. The expression of hMSH2 could be used as a potential marker for distinguishing benign insulinoma from malignant ones.