The efficacy of tofacitinib combined with leflunomide in the treatment of rheumatoid arthritis and its effect on serum janus kinase/signal transduction and activator of transcription signaling pathway-related proteins and matrix metalloproteinases levels / 中华风湿病学杂志

Objective:To investigate the clinical efficacy of Tofacitinib combined with leflunomide in the treatment of rheumatoid arthritis (RA) and its effects on Janus Kinase (JAK)/signal transduction and activator of transcription (STAT) signaling pathway-related proteins and Matrix metalloproteinase levels in serum of patients with RA.Methods:This was a prospective case-control study. A total of 80 patients with RA in our hospital from March 2020 to September 2021 were included into the study. They were divided into observation group and control group by random number table method, with 40 cases in each group. The patients in observation group were treated with Tofacitinib combined with leflunomide, while the patients in control group were treated with leflunomide alone. After 12 weeks of continuous treatment, the curative effect of the two groups was observed. Typical clinical manifestation [including Visual analogue scale (VAS) score of joint pain, number of tenderness joints, number of swollen joints, time of morning stiffness], disease activity score uses 28 joint counts (DAS28) scores, the MOS item short from health survey (SF-36) total scores and serum JAK3, STAT3, interleukin (IL)-6, IL-17 and matrix metalloproteinase (MMPs) levels were compared between the two groups before and after treatment. The adverse effects of the two groups were also analyzed. Chi-square test, paired sample t test, independent sample t test and Fisher exact probability method were used for statistical analysis. Results:After 4, 8 and 12 weeks of treatment, the american college of rheumatology (ACR)20 compliance rates in the observation group were 37.5%(15/40), 62.5%(25/40) and 80.0%(32/40), respectively, which were significantly higher than those in the control group at the same period [17.5%(7/40), 37.5%(15/40), 57.5%(23/40); χ2=4.01, P=0.045; χ2=5.00, P=0.025; χ2=4.71, P=0.030]. After 8 and 12 weeks of treatment, the ACR50 compliance rates in the observation group were [35.0%(14/40) and 47.5%(19/40), respectively, which were significantly higher than those in the control group at the same period 15.0%(6/40) and 20.0%(8/40), χ2=4.27, P=0.039; χ2=6.76, P=0.009]. After treatment, the joint pain VAS score, number of tenderness joints, number of swollen joints, DAS28 scores and SF-36 total scores in the observation group were lower than those in the control group( t=5.55, P<0.001; t=9.98, P<0.001; t=11.77, P<0.001; t=4.50, P<0.001; t=5.28, P<0.001), and time of morning stiffness was shorter than that in the control group ( t=4.76, P<0.001). After treatment, The serum levels of JAK3, STAT3, IL-6, IL-17, MMP-3, MMP-9 and MMP-13 in the obser vation group were (2 354±476) pg/ml, (1.04±0.17) ng/ml, (12.4±2.8) pg/ml, (30±5) pg/mL, (65±14) μg/L, (76±12) μg/L, (11.5±1.8) μg/L, which were lower than those in control group [(2 715±584) pg/ml (1.22±0.29) ng/mL, (16.8±3.6) pg/ml, (40±7) pg/ml, (98±15) μg/L, (123±20) μg/L, (14.9±2.8) μg/L, t=3.03, P<0.05; t=3.39, P<0.05; t=6.10, P<0.05; t=7.35, P<0.05; t=10.17, P<0.05; t=12.74, P<0.05; t=6.46, P<0.05]. The adverse reaction rate of the observation group [35.0%(14/40)] had no statistically significant difference compared with that in the control group [27.5%(11/40)]( χ2=0.52, P=0.469). Conclusion:The overall effect of Tofacitinib com bined with leflunomide in the treatment of RA is satisfactory and it is a safe and effective way to improve the clinical manifestation, disease activity and quality of life of patients with RA. The effect may be related to the significant down-regulation of the expression levels of Serum JAK/STAT signaling pathway-related Proteins and MMPs.

Effect of Anchang Decoction on Inflammation and Immunity in Rats with Ulcerative Colitis Based on miRNA-146a/JAK/STAT/SOCS-3 Signal Pathway / 中国实验方剂学杂志

Objective:To study the anti-inflammatory effects of low， middle， and high doses of Anchang decoction on ulcerative colitis in SD rats， and also explore the possible mechanism of Anchang decoction in the prevention and treatment of ulcerative colitis through the effect of different doses on miRNA-146a/non-receptor tyrosine protein kinase（JAK）/signal transduction and activator of transcription 3（STAT3）/cytokine signaling protein-3（SOCS-3） signal pathway and its downstream proteins. Method:The experimental rats were divided into control group ， model group ， mesalazine group（1 g·kg^-1） and Anchang decoction low（6 g·kg^-1）， middle（12 g·kg^-1）and high dose groups（24 g·kg^-1）， with 10 rats in each group. Except for the control group， 2，4，6-trinitrobenzenesulfonic acid （TNBS）/ethanol enema was used in all the other groups to establish a rat model of ulcerative colitis for 14 days respectively. The general changes of the mental state， stool traits， hair and other general conditions of the rats were observed， and score was graded with reference to the disease activity index （DAI） table. The pathological changes of colon tissue of rats in each group were observed by hematoxylin-eosin （HE） staining. The levels of serum tumor necrosis factor-<italic>α</italic>（TNF-<italic>α</italic>）， interleukin-10（IL-10）， interleukin-17（IL-17）， interleukin-1<italic>β</italic>（IL-1<italic>β</italic>）and interleukin-6（IL-6）were detected by enzyme linked immunosorbent assay （ELISA）. The expression levels of JAK2， phosphorylation STAT3 （p-STAT3）， STAT3 and inhibitor of SOCS-3 in colon tissue were detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the expression of JAK2， p-STAT3， STAT3， SOCS-3 mRNA in rat colon and miRNA-146a in rat plasma. Result:Compared with the normal group， the expression of JAK2， p-STAT3， STAT3 protein and the expression of JAK2， p-STAT3 and STAT3 mRNA in the model group increased （<italic>P</italic><0.05）， and the relative expression of miRNA-146a， SOCS-3 mRNA and SOCS-3 protein decreased in the model group （<italic>P</italic><0.05）. Compared with the model group， the mental state， food intake， coat color， etc. of rats in the administration groups were significantly improved， the DAI score was significantly reduced （<italic>P</italic><0.05）， the colonic ulcer tissues of rats in the administration groups were improved significantly， the expression levels of JAK2， p-STAT3， STAT3 protein and the expression of JAK2， p-STAT3 and STAT3 mRNA in the colon tissue of the administration groups were decreased （<italic>P</italic><0.05）， and the relative expression levels of miRNA-146a， SOCS-3 mRNA and SOCS-3 protein were increased in the administration groups （<italic>P</italic><0.05）. Conclusion:Anchang decoction can alleviate ulcerative colitis and reduce the activation of inflammatory factors by affecting the expression of genes and proteins related to miRNA-146a/JAK/STAT/SOCS-3 signal transduction pathway.

Glaucocalyxin A regulates STAT3 signaling pathway through influencing IL-6 expression in HCCLM3 cells / 中华微生物学和免疫学杂志

Objective To investigate the effects of glaucocalyxin A ( GLA) on the expression of signal transduction and activator of transcription 3 (STAT3) signaling pathway in HCCLM3 cells for better understand-ing the role of GLA in tumor metastasis. Methods HCCLM3 cells were stimulated by different concentrations (0. 1 ng/ml, 1 ng/ml, 10 ng/ml, 100 ng/ml and 1000 ng/ml) of GLA for 48 h. Expression of IL-6, STAT3/pSTAT3Tyr705 and MMP9 at mRNA and protein levels were detected by qRT-PCR and Western blot, respectively. Transwell invasion and migration assays were performed to analyze cell invasion and migration. ELISA was used to measure the concentrations of IL-6 in culture media. Results Expression of IL-6 at mRNA and protein lev-els, STAT3Tyr705 phosphorylation, and the migration and invasion of HCCLM3 cells were significantly enhanced after stimulation of HCCLM3 cells with 0. 1 ng/ml of GLA, but inhibited by GLA at the concentration of 1000 ng/ml. Moreover, more IL-6 was secreted out of the HCCLM3 cells treated with 0. 1 ng/ml of GLA. Conclu-sions GLA might affect the metastasis of HCCLM3 cells through regulating the expression of IL-6 and the phos-phorylation of STAT3Tyr705. These regulatory effects were closely related to the concentration of GLA.

Effect of insulin-like growth factor 1 on phagocytosis and production of interleukin-10 in murine alveolar epithelial cells / 上海交通大学学报（医学版）

Objective • To investigate the effect of insulin-like growth factor 1 (IGF-1) on phagocytosis and production of interleukin-10 (IL-10) in the murine alveolar epithelial cell line MLE-12. Methods • After treatment with IGF-1 for 48 h, MLE-12 cells were incubated with fluorescent microspheres for 2 h in the presence or absence of wortmannin (phosphatidylinositol-3-kinase inhibitor). Flow cytometry was then used to assess cell phagocytosis of fluorescent microspheres. Enzyme linked immunosorbent assay (ELISA) was used to detect the content of IL-10 in MLE-12 cells culture supernatant stimulated by IGF-1 for 24 h. After pretreatment with IGF-1 for 2 h, MLE-12 cells were stimulated by lipopolysaccharides (LPS) for 24 h, and the expression of phosphorylated signal transduction and activator of transcription 3 (p-STAT3) in cytoplasm was detected by Western blotting.Results • With the increase of IGF-1 stimulation concentration, the ability of MLE-12 cells to phagocytose fluorescent microspheres increased, and the ability to phagocytose fluorescent microspheres reached the peak in the presence of IGF-1 at 50 ng/mL. However, the ability of IGF-1 to phagocytose fluorescent microspheres was completely blocked by wortmannin in MLE-12 cells. IGF-1 promoted IL-10 secretion and inhibited LPS-induced enhancement of STAT3 activation in MLE-12 cells. Conclusion • IGF-1 promotes phagocytosis of MLE-12 cells via the PI3K/protein kinase B (Akt) pathway and exhibits anti-inflammatory properties.

Effect of GP130 small molecular inhibitor SC144 on renal interstitial fibrosis in mice with unilateral ureteral obstruction / 上海交通大学学报（医学版）

Objective·To investigate the effect of glycoprotein 130 (GP130) inhibitor SC144 on extracellular matrix accumulation and JAK2/STAT3 signaling pathway in unilateral ureteral obstruction (UUO) mouse model, and explore its mechanism. Methods·Eighteen female BALB/c mice were randomly divided into 3 groups i.e. sham group, UUO group and SC144 group. All mice were sacrificed at day 14 and kidneys were harvested for further analysis. The changes of renal tissue morphology and pathology were observed by H-E and Masson staining. The expression of α-smooth muscle actin (α-SMA) and infiltration of macrophage cells were assayed by immunohistochemical staining. The levels of collagen-I, collagen-IV, monocyte chemoattractant protein-1(MCP-1),transforming growth factor-β(TGF-β)mRNA were analyzed by real-time PCR.The activation of JAK2 and STAT3 was measured by Western blotting. Results·There was a trend toward decreased renal tubular lesion and renal interstitial fibrosis in SC144 group (H-E, P=0.052;Masson,P=0.063).SC144 significantly inhibited the levels of α-SMA,type I/type IV collagen and TGF-β mRNA(all P<0.05).Compared with UUO group, the phosphorylation levels of JAK2 and STAT3 were significantly decreased in SC144 group (both P<0.05). Conclusion·The treatment of UUO mouse model with SC144 can inhibit the activation of α-SMA, attenuate the phosphorylation of STAT3, reduce extracellular matrix protein deposition following injury and renal tubular epithelial-mesenchymal transition(EMT)via JAK2/STAT3 signaling pathway,indicating its potential in attenuating interstitial fibrosis in obstructive nephropathy.

Effect of GP130 small molecular inhibitor SC144 on renal interstitial fibrosis in mice with unilateral ureteral obstruction / 上海交通大学学报（医学版）

Objective: To investigate the effect of glycoprotein 130 (GP130) inhibitor SC144 on extracellular matrix accumulation and JAK2/STAT3 signaling pathway in unilateral ureteral obstruction (UUO) mouse model, and explore its mechanism. Methods: Eighteen female BALB/c mice were randomly divided into 3 groups i.e. sham group, UUO group and SC144 group. All mice were sacrificed at day 14 and kidneys were harvested for further analysis. The changes of renal tissue morphology and pathology were observed by H-E and Masson staining. The expression of α-smooth muscle actin (α-SMA) and infiltration of macrophage cells were assayed by immunohistochemical staining. The levels of collagen-I, collagen-IV, monocyte chemoattractant protein-1 (MCP-1), transforming growth factor-β (TGF-β) mRNA were analyzed by real-time PCR. The activation of JAK2 and STAT3 was measured by Western blotting. Results: There was a trend toward decreased renal tubular lesion and renal interstitial fibrosis in SC144 group (H-E, P=0.052; Masson, P=0.063). SC144 significantly inhibited the levels of α-SMA, type I/type IV collagen and TGF-β mRNA (all P<0.05). Compared with UUO group, the phosphorylation levels of JAK2 and STAT3 were significantly decreased in SC144 group (both P<0.05). Conclusion: The treatment of UUO mouse model with SC144 can inhibit the activation of α-SMA, attenuate the phosphorylation of STAT3, reduce extracellular matrix protein deposition following injury and renal tubular epithelial-mesenchymal transition (EMT) via JAK2/STAT3 signaling pathway, indicating its potential in attenuating interstitial fibrosis in obstructive nephropathy.