ABSTRACT
Objective:To explore the expression and clinical significance of Janus protein tyrosine kinase/signal transducer and activator of transcription (JAK3/STAT5) signaling pathway in peripheral blood mononuclear cells (PBMCs) of patients with primary gouty arthritis (GA).Methods:Peripheral blood, clinical data and laboratory tests were collected from 50 patients with acute gout (AG), 50 patients with intermittent gout (IG) and 50 healthy controls (HC). Quantitative real-time-polymerase chain reaction (RT-qPCR) was used to detect mRNA expression level of JAK3/STAT5 related genes (JAK3, STAT5a, STAT5b). Enzyme linked immune sorbent assay (ELISA) was used to detect interleukin-2 (IL-2) concentration in subject′s plasma. Measurement data among the three groups that was in accordance with normal distribution was analyzed by one-way analysis of variance, pairwise comparisons using LSD, non-normal distribution data was analyzed by Mann-Whitney test or Kruskal-Wallis H test, and correlation analysis between variables was analyzed using Spearman correlation analysis. Results:① The mRNA expression levels of JAK3, STAT5a and STAT5b were significantly different among the three groups ( F=50.13, P<0.01; F=7.573, P=0.000 7; F=12.14, P<0.01), of which the JAK3 mRNA expression level in the HC group [(606±65)×10 -4] was significantly higher than that in the AG group [(103±13)×10 -4] and IG group [(114±24)×10 -4], and the difference was statistically significant (both P<0.01), while the STAT5a mRNA expression level in the AG group [(89±9)×10 -4] was significantly higher than that in the IG group [(59±4)×10 -4] and HC group [(61±4)×10 -4], and the difference was statistically significant ( P=0.002, P=0.003 9), and the expression level of STAT5b mRNA in HC group [(60±5)×10 -4] was significantly lower than that in AG group [(95±7)×10 -4] and IG group [(98±7)×10 -4], and the difference was statistically significant ( P=0.000 2, P<0.000 1). ② The difference of IL-2 concentration in plasma among the three groups was statistically significant ( F=22.87, P<0.01), and the serum IL-2 concentration in the AG group [(87.9±8.4) pg/ml] was significantly higher than that in the IG group [(32±4) pg/ml] and HC group [(44±4) pg/ml], and the difference was statistically significant (both P<0.01). ③ Spearman correlation analysis showed that the mRNA expression of STAT5a and STAT5b was positively correlated with the absolute value of neutrophils in patients with gout ( r=0.282, P<0.05; r=0.257, P<0.05). Conclusion:The IL-2/JAK3/STAT5 signaling pathway is involved in the occurrence and development of gout, suggesting that this pathway may play a key role in the pathogenesis of gout.
ABSTRACT
Objective::To observe the effect of Qingzao Jiufei Tang on apoptosis of lung cancer, Janus protein tyrosine kinase 2/signal transducers and transcriptional activator protein 3 (JAK2/STAT3) signaling pathway, as well as the expressions of downstream apoptosis-related proteins Bcl-2-associated X (Bax) and Cyclin D1. Method::Totally 50 male C57BL/6J mice were randomly divided into five groups: chemotherapy group (CTX), model group, high-dose Qingzao Jiufei Tang group, middle-dose Qingzao Jiufei Tang group and low-dose Qingzao Jiufei Tang group, with 10 mice in each group. The model of lung cancer was established by injecting Lewis lung cancer cells into the right axillary of mice. High-dose, middle-dose and low-dose Qingzao Jiufei Tang groups were orally given drugs (11, 5.5, 2.75 g·kg-1·d-1) two weeks before the modeling. Chemotherapy group was administered intraperitoneally at a dose of 50 mg·kg-1·(2 d)-1, while model group was administered intragastrically with the equal volume of normal saline. After inoculation, the mice in each group were continued to be administered. Two weeks later, the mice in each group were killed, and the tumors were collected. Then the JAK2 protein phosphorylation level was detected by immunohistochemistry (IHC). STAT3, Bax and Cyclin D1 protein expression levels were detected by Western blot, and apoptosis of lung cancer cells was observed by transmission electron microscopy. Result::Compared with model group, the phosphorylation levels of JAK2 and STAT3 protein in lung cancer cells were significantly decreased, the expression of Bax protein was significantly increased, and the expression of Cyclin D1 protein was significantly decreased in high-dose Qingzao Jiufei Tang group, middle-dose Qingzao Jiufei Tang group and chemotherapy group, with statistically significant differences (P<0.05, P<0.01). The results of transmission electron microscopy showed significant apoptotic phenomena in high-dose Qingzao Jiufei Tang group, middle-dose Qingzao Jiufei Tang group, low-dose Qingzao Jiufei Tang group and chemotherapy group compared with the model group. Conclusion::Qingzao Jiufei Tang had an obvious effect in promoting the apoptosis of lung cancer cells. Its mechanism may be related to the inhibition of the phosphorylation of JAK2 and STAT3 protein, the promotion of its downstream Bax protein expression and the inhibition of its downstream Cyclin D1 protein expression.
ABSTRACT
Objective To study the activation of Janus protein tyrosine kinase (JAK)/signal transducer and activator of transcription 1 (STAT1) signaling pathway and its inhibitor-signal transducer and activator of transcription-1(SOCS-1) in patients with systemic lupus erythematosus. Methods A total of 45 patients with active systemic lupus erythematosus (SLE) and 30 healthy controls were randomly selected. Western blot was performed to measure the expression of Stat1 protein and phospho-Stat1 protein (an activated form of Stat1 protein) in the monocytes after stimulation with recombinant high mobility group box1 (rHMGB1) at various time points. Expression of Stat1 protein in the skin or lesional skin was also detected. Phasic expressions of SOCS-1 mRNA in the monocytes after rHMGB1 stimulation were detected by real-time reverse transcription-polymerase chain reaction. SOCS-1 gene expression in the skin or lesional skin was also detected. Results The expression level of Stat1 proteins in the monocytes from patients with SLE was higher than that from healthy controls (t=9.16,P＜0.01) and positively correlated with SLE disease activity index (SLEDAI) (r=0.59,P＜0.01). Expression of phospho-Stat1 in the monocytes from SLE patients was time-dependently upregulated after stimulation with rHMGB1 at various time points, while expression of SOCS-1 mRNA remained unchanged(all P＞0.05). Expressions of phospho-Stat1 protein and SOCS-1 mRNA in the monocytes from healthy controls were increased transiently after stimulation with rHMGB1(all P＜0.05). Both expressions of phospho-Stat1 protein and SOCS-1 gene in the lesional skin from patients with SLE were upregulated compared with those in normal skin from healthy controls (all P＜0.01). Conclusion There are hyperactivation of JAK-STAT1 signaling pathway and negative feedback down-regulation of SOCS-1 in patients with systemic lupus erythematosus. HMGB-1 may be partly involved in the pathogenesis of SLE by the abnormal mediating function of JAK-STAT1 signal transduction pathway.