Gegen Qinlian decoction ameliorates TNBS-induced ulcerative colitis by regulating Th2/Th1 and Tregs/Th17 cells balance, inhibiting NLRP3 inflammasome activation and reshaping gut microbiota.

ETHNOPHARMACOLOGICAL RELEVANCE: Chinese herbal medicine Gegen Qinlian Decoction (GQD) has been clinically shown to be an effective treatment of ulcerative colitis (UC) in China. However, the underlying mechanism of GQD's anti-ulcerative colitis properties and its effect on gut microbiota still deserve further exploration. AIM OF THE STUDY: This study observed the regulatory effects of GQD on Th2/Th1 and Tregs/Th17 cells balance, the NOD-like receptor family pyrin domain containing 3 (NLRP3) infammasome and gut microbiota in TNBS-induced UC in BALB/c mice. MATERIALS AND METHODS: 61 main chemical compounds in the GQD were determined by UPLC-Q-TOF/MS. The UC BALB/c model was established by intrarectal administration of trinitrobenzene sulfonic acid (TNBS), and GQD was orally administered at low and high dosages of 2.96 and 11.83 g/kg/day, respectively. The anti-inflammatory effects of GQD for ulcerative colitis were evaluated by survival rate, body weight, disease activity index (DAI) score, colonic weight and index, spleen index, hematoxylin-eosin (HE) staining and histopathological scores. Flow cytometry was used to detect the percentage of CD4, Th1, Th2, Th17 and Tregs cells. The levels of Th1-/Th2-/Th17-/Tregs-related inflammatory cytokines and additional proinflammatory cytokines (IL-1ß, IL-18) were detected by CBA, ELISA, and RT-PCR. The expressions of GATA3, T-bet, NLRP3, Caspase-1, IL-Iß, Occludin and Zonula occludens-1 (ZO-1) on colon tissues were detected by Western blot and RT-PCR. Transcriptome sequencing was performed using colon tissue and 16S rRNA gene sequencing was performed on intestinal contents. Fecal microbiota transplantation (FMT) was employed to assess the contribution of intestinal microbiota and its correlation with CD4 T cells and the NLRP3 inflammasome. RESULTS: GQD increased the survival rate of TNBS-induced UC in BALB/c mice, and significantly improved their body weight, DAI score, colonic weight and index, spleen index, and histological characteristics. The intestinal barrier dysfunction was repaired after GQD administration through promoting the expression of tight junction proteins (Occludin and ZO-1). GQD restored the balance of Th2/Th1 and Tregs/Th17 cells immune response of colitis mice, primarily inhibiting the increase in Th2/Th1 ratio and their transcription factor production (GATA3 and T-bet). Morever, GQD changed the secretion of Th1-/Th2-/Th17-/Tregs-related cytokines (IL-2, IL-12, IL-5, IL-13, IL-6, IL-10, and IL-17A) and reduced the expressions of IL-1ß, IL-18. Transcriptome results suggested that GQD could also remodel the immune inflammatory response of colitis by inhibiting NOD-like receptor signaling pathway, and Western blot, immunohistochemistry and RT-PCR further revealed that GQD exerted anti-inflammatory effects by inhibiting the NLRP3 inflammasome, such as down-regulating the expression of NLRP3, Caspase-1 and IL-1ß. More interestingly, GQD regulated gut microbiota dysbiosis, suppressed the overgrowth of conditional pathogenic gut bacteria like Helicobacter, Proteobacteria, and Mucispirillum, while the probiotic gut microbiota, such as Lactobacillus, Muribaculaceae, Ruminiclostridium_6, Akkermansia, and Ruminococcaceae_unclassified were increased. We further confirmed that GQD-treated gut microbiota was sufficient to relieve TNBS-induced colitis by FMT, involving the modulation of Th2/Th1 and Tregs/Th17 balance, inhibition of NLRP3 inflammasome activation, and enhancement of colonic barrier function. CONCLUSIONS: GQD might alleviate TNBS-induced UC via regulating Th2/Th1 and Tregs/Th17 cells Balance, inhibiting NLRP3 inflammasome and reshaping gut microbiota, which may provide a novel strategy for patients with colitis.

Jiedu Quyu Decoction mitigates monocrotaline-induced right-sided heart failure associated with pulmonary artery hypertension by inhibiting NLRP3 inflammasome in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Right-side heart failure could accelerate mortality in patients of pulmonary hypertension, Jiedu Quyu Decoction (JDQYF) was used to manage pulmonary hypertension, but its right-sided heart protective effect associated with pulmonary artery hypertension is still unclear. AIM OF THE STUDY: Here, we evaluated the therapeutic effect of JDQYF on monocrotaline-induced right-sided heart failure associated with pulmonary arterial hypertension in Sprague-Dawley (SD) rats and investigated the potential mechanism of action. MATERIALS AND METHODS: The main chemical components of JDQYF were detected and analyzed using ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry. The effects of JDQYF were investigated using a rat model of monocrotaline-induced right-sided heart failure associated with pulmonary arterial hypertension. We assessed the morphology of cardiac tissue using histopathology and the structure and function of the right heart using echocardiography. The biomarkers of heart failure, atrial natriuretic peptide and B-type natriuretic peptide, as well as serum pro-inflammatory markers, interleukin (IL)-1ß, and IL-18, were measured by enzyme-linked immunosorbent assay (ELISA). Furthermore, the mRNA and protein expression levels of NLRP3 (NOD-, LRR-, and pyrin domain-containing 3), capase-1, IL-1ß, and IL-18 in the right heart tissue were examined by real-time quantitative reverse transcription PCR and western blotting. RESULTS: JDQYF improved ventricular function, alleviated pathological lesions in the right cardiac tissue, reduced the expression levels of biomarkers of heart failure and serum pro-inflammatory factors (IL-1ß and IL-18), and downregulated the mRNA and protein expression levels of NLRP3, caspase-1, IL-1ß, and IL-18 in the right cardiac tissue. CONCLUSIONS: JDQYF possesses cardioprotective effect against right heart failure induced by pulmonary arterial hypertension, possibly owing to reduction of cardiac inflammation through the inhibition of NLRP3 inflammasome activation.

Correlation Between Plasma NLRP3, IL-1ß, and IL-18 and Diabetic Nephropathy in Patients With Type 2 Diabetes.

Context: Diabetic nephropathy (DN) is a common microvascular complication in diabetic patients. The pathogenesis of DN is complex. Inflammatory response may play a key role as a common downstream pathway. Objective: The study intended to explore the relationship between the levels of plasma nucleotide-binding oligomeric domain-like receptor protein 3 (NLRP3 inflammasome), interleukin-1ß (IL-1ß), and IL-18 and the progression of type 2 diabetic nephropathy to clarify their relationship with type 2 diabetes mellitus (T2DM) and to provide evidence for clinical treatment. Design: The research team performed a controlled observational study. Setting: The study took place at Baoding No. 1 Central Hospital in Baoding, Hebei, China. Participants: Participants were 153 patients with T2DM who received treatment at the hospital between October 2020 and October 2021. The research team allocated 30 participants without evidence of DN to the control group. Based on the DN stage, the team assigned the 123 remaining participants to one of five observation groups: (1) 32 participants with stage 1 DN to the DN1 group, (2) 31 participants with stage 2 DN to the DN2 group, (3) 30 participants with stage 3 DN to the DN3 group, (4) 30 participants with stage 4 DN to the DN4 group, and (5) 29 participants with stage 5 DN to the DN5 group. Outcome Measures: The research team measured participants' levels of "nucleotide binding oligomeric domain-like receptor protein 3" (NLRP3), interleukin-1 beta (IL-1ß), and IL-18 and used the Spearman rank correlation analysis to determine the correlation between those levels and the DN stages. Results: The levels of NLRP3 , IL-1ß and IL-18 in all the five observation groups were significantly higher than those in the control group (all P < .01). The levels were also significantly higher: (1) in the DN2, DN3, DN4, and DN5 groups than those in the DN1 group (all P < .01); (2) in the DN3, DN4, and DN5 groups than those in the DN2 group (all P < .01); (3) in the DN4 and DN5 groups than those in the DN3 group (all P < .01); and (4) in the DN5 groups than those in the DN4 group (all P < .01). The Spearman rank correlation analysis showed that the NLRP3, IL-1ß, and IL-18 levels were significantly positively correlated with the DN stage (P = .01). Conclusions: NLRP3, IL-1ß and IL-18 played an important role in the progression of T2DM, and their levels increased with the aggravation of DN. Therefore, the plasma levels of NLRP3, IL-1ß and IL-18 can be useful as indicators of the occurrence and development of DN and can provide clinical guidance for the early diagnosis of DN and for the determination and adjustment of treatment plans.

Tibetan medicine Qi-Sai-Er-Sang-Dang-Song Decoction inhibits TNF-α-induced rheumatoid arthritis in human fibroblast-like synoviocytes via regulating NOTCH1/NF-κB/NLRP3 pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Qi-Sai-Er-Sang-Dang-Song Decoction (QSD, ཆུ་སེར་སེང་ལྡེང་སུམ་ཐང་།), a Tibetan classical herbal formula, is commonly used in Tibetan hospital preparation for the treatment of rheumatoid arthritis (RA). Its efficacy is to relieve inflammation, dispel cold, remove dampness, and alleviate pain. However, its anti-RA mechanism is still unclear. AIM OF THE STUDY: This study aimed to investigate the effect of QSD on rheumatoid arthritis and explore its anti-inflammatory mechanism against human fibroblast-like synoviocytes (HFLSs) by regulating the notch family of receptors (NOTCH1)/Nuclear factor-κB (NF-κB)/nucleotide-binding (NLRP3) pathway. MATERIALS AND METHODS: We used ultra-performance liquid chromatography coupled with Q-TOF mass spectrometry (UPLC-Q-TOF-MS) to identify the chemical composition of QSD. Then, HFLSs were exposed to drug-containing serum. The effect of QSD drug-containing serum on HFLS viability was detected using the cell counting kit-8 (CCK-8) assay. Next, we explored the anti-inflammatory effect of QSD using enzyme-linked immunosorbent assay (ELISA) for inflammatory factors, such as interleukin-18 (IL-18), interleukin-1ß (IL-1ß), and interleukin-6 (IL-6). The expression of NOTCH-related proteins, a member of the NOTCH1, Cleaved NOTCH1, hairy and enhancer of split-1 (HES-1), NF-κB p65, NF-κB pp65, NLRP3, and delta-like 1 (DLL-1), was examined using western blotting. Furthermore, the relative mRNA expression levels of NOTCH1, NF-κB p65, NLRP3, DLL-1, and HES-1 were detected using real-time quantitative (RT-qPCR). To explore the mechanism underlying the anti-RA effect of QSD, we the used the NOTCH signaling pathway inhibitor LY411575 and transfection with a NOTCH1 siRNA. In addition, we employed immunofluorescence to determine the expression of HES-1 and NF-κB p65 in vitro. RESULT: Our results revealed that QSD ameliorated inflammation in HFLSs. Compared with the model group, the QSD drug-containing serum group had obviously down-regulated levels of IL-18, IL-1ß, and IL-6. Consistently, the CCK-8 results showed that the QSD drug-containing serum had no obvious toxicity towards HFLSs. Moreover, both LY411575 and siNOTCH1, QSD could reduce NOTCH1, NLRP3, and HES-1 protein expression levels, and LY411575 could significantly inhibit the expression levels of NF-κB p65, NF-κB pp65, and Cleaved NOTCH1 (p < 0.05). siNOTCH1 could also suppress the expression of DLL-1. The RT-qPCR results indicated that QSD could downregulate the relative mRNA expression levels of NOTCH1, NF-κB p65, NLRP3, DLL-1, and HES-1 in HFLSs (p < 0.05). In the immunofluorescence experiment, the fluorescence intensities of HES-1 and NF-κB p65 in HFLSs were found to decrease after exposure to QSD drug-containing serum (p < 0.05). Ultimately, 44 chemical components were detected in QSD using UPLC-Q-TOF-MS. CONCLUSION: This study reveals that the QSD can markedly ameliorate inflammation induced by TNF-α on HFLS. The effect of QSD on HFLS may be exerted by inhibition of the NOTCH1/NF-κB/NLRP3 signaling pathway.

Effects of n-butanol extract of Pulsatilla decoction on the NLRP3 inflammasome in macrophages infected with Candida albicans.

ETHNOPHARMACOLOGICAL RELEVANCE: Pulsatilla decoction is a traditional Chinese medicine from Shang Han Lun that has been reported to have therapeutic efficacy in vulvovaginal candidiasis (VVC), and is a growth inhibitor of Candida albicans (C. albicans) in vitro, the causative agent of VVC. AIM OF THE STUDY: In previous studies, Pulsatilla decoction has shown therapeutic benefits for VVC. With further chemical extraction of the decoction, the n-butanol extract (of Pulsatilla decoction; BEPD) was most effective against C. albicans and therapeutic for VVC. The mechanism, however, has not been elucidated. The regulation of NOD-like receptor protein 3 (NLRP3) inflammasome has recently been demonstrated as a critical component of the inflammasome complex that initiates the vaginal inflammatory response. Therefore, the effect of BEPD on NLRP3 associated with VVC was investigated. MATERIALS AND METHODS: Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used for detecting the principal compounds of BEPD (Anemoside B4, Esculin, esculetin, Epiberberine, Berberine, Jatrorrhizine and Phellodendrine). BEPD-containing serum is prepared by intragastric administration of BEPD (4.6875 g/kg for seven days) in rats. PMA-induced THP-1 cells were challenged with C. albicans. The expression of CD68 to identify macrophages was examined by flow cytometry, the viability of THP-1 cells were assessed by CCK8 assay, the release of lactate dehydrogenase (LDH) was detected by LDH kit, and the secretion levels of IL-1ß and IL-18 were evaluated through enzyme-linked immunosorbent assay (ELISA), the levels of NLRP3 were quantified by immunofluorescence, the levels of reactive oxygen species (ROS) were measured by ROS kit, and the expression of Dectin-1, Syk, TLR2, TLR4, MyD88, NF-κB, NLRP3, Caspase-1, and ASC proteins were detected by Western blot. RESULTS: After administration of BEPD-containing serum, the levels of IL-1ß, IL-18 and LDH released from macrophages were reduced in the BEPD-containing serum group compared to the model group. In addition, BEPD-containing serum inhibited the expression of ROS in macrophages, suppressed the expression of NLRP3 and inhibited the expression of TLRs/MyD88 and Dectin-1/Syk signaling pathway-related proteins. CONCLUSIONS: BEPD suppressed the NLRP3 inflammasome and related signaling pathways in macrophages infected with C. albicans in vitro, thereby providing insight into the mechanism of BEPD action on VVC.

Xianglian Pill attenuates ulcerative colitis through TLR4/MyD88/NF-κB signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Xianglian Pill (XLP) is a classical Chinese medicine prescription applied for controlling ulcerative colitis (UC). Whereas, the underlying mechanism remains unclear. AIM OF THE STUDY: The present work was aimed to investigate the mechanism of XLP in dextran sulfate sodium (DSS)-induced UC via the Toll Like Receptor 4 (TLR4)/Myeloid Differentiation factor 88 (MyD88)/Nuclear Factor kappa-B (NF-κB) signaling in mice. MATERIALS AND METHODS: The major components of XLP were detected by high-performance liquid chromatography-diode array detection (HPLC-DAD). The ulcerative colitis model was induced by DSS in mice. 5-Amino Salicylic Acid (5-ASA) group and XLP group were intragastrically treated. Disease activity index (DAI) and colon length were monitored and hematoxylin-eosin (HE) staining was conducted. Gasdermin D (GSDMD)-N and TLR4 expressions in colon tissues were visualized by immunofluorescence. TLR4 mRNA was measured by Real Time Quantitative PCR (RT-qPCR). The expressions of NOD-like receptor thermal protein domain associated protein 3 (NLRP3), active-caspase-1, GSDMD-N, TLR4, MYD88, NF-κB, p-NF-κB, and the ubiquitination of TLR4 in colon tissues were detected by Western blot. Myeloperoxidase (MPO) enzyme activity was examined and serum inflammatory factors Interleukin (IL)-1ß, IL-6, Tumor Necrosis Factor-α (TNF-α), and IL-18 were determined by Enzyme-linked Immunosorbent Assay (ELISA). TLR4-/- mice were applied for verifying the mechanism of XLP attenuated DSS symptoms. RESULTS: The XLP treatment extended colon length, reduced DAI, and attenuated histopathological alteration in DSS-induced mice. XLP administration suppressed MPO activity and reduced the content of IL-1ß, IL-6, TNF-α and IL-18 in serum. XLP also inhibited the expression levels of GSDMD-N, TLR4, NLRP3, active-caspase-1, MyD88, p-NF-κB/NF-κB in colon tissues of DSS-induced mice. TLR4-/- mice proved that TLR4 was involved in XLP-mediated beneficial effect on DSS-induced ulcerative colitis. CONCLUSIONS: XLP might treat ulcerative colitis by regulating the TLR4/MyD88/NF-κB signaling pathway.

Qing-Chang-Hua-Shi granule ameliorates DSS-induced colitis by activating NLRP6 signaling and regulating Th17/Treg balance.

BACKGROUND: Chinese herbal medicine Qing-Chang-Hua-Shi granule (QCHS) is widely used to treat ulcerative colitis in China. However, the molecular mechanisms of QCHS remains largely unknown. PURPOSE: To assess the therapeutic effects of QCHS on colitis and to reveal its mechanisms of action. METHODS: The main components of QCHS were identified using a UHPLC-QTOF-MS method and the efficacy of QCHS was evaluated using an DSS-induced mice model. The inflammatory responses and mucosal integrity in colon were comprehensively assessed. Flow cytometry was used to analysis the proportion of Th17 and Treg cells. Detect the signal transduction of the NOD-like receptor family pyrin domain containing 6 (NLRP6) both in vitro and in vivo. Furthermore, siNLRP6 transfection was used to validate the functional targets of QCHS. RESULTS: QCHS treatment significantly alleviated colitis in mice by improving symptoms and pathological damage. Moreover, QCHS treatment suppressed the inflammatory response and preserved the integrity of colon tissue. Most importantly, QCHS balanced the Th17/Treg response of UC mice. Mechanistically, by activating NLRP6 inflammasome pathway, QCHS regulated the maturation of interleukin (IL)-1ß and IL-18 to affect inflammation and drive Th17 cell differentiation. CONCLUSIONS: The effect of QCHS on UC mice is dose-dependent, with high-dose QCHS being superior to 5-Aminosalicylic acid (200 mg/kg/day). QCHS acts through the NLRP6 signaling pathway to modulate Th17/Treg balance, resulting in the protective effects against colitis. This study investigated the relevant pharmacological mechanisms of QCHS, providing further evidence for the application of QCHS in UC treatment.

Anti-inflammatory potential of stevia residue extract against uric acid-associated renal injury in mice.

Abnormal uric acid level result in the development of hyperuricemia and hallmark of various diseases, including renal injury, gout, cardiovascular disorders, and non-alcoholic fatty liver. This study was designed to explore the anti-inflammatory potential of stevia residue extract (STR) against hyperuricemia-associated renal injury in mice. The results revealed that STR at dosages of 150 and 300 mg/kg bw and allopurinol markedly modulated serum uric acid, blood urea nitrogen, and creatinine in hyperuricemic mice. Serum and renal cytokine levels (IL-18, IL-6, IL-1Β, and TNF-α) were also restored by STR treatments. Furthermore, mRNA and immunohistochemistry (IHC) analysis revealed that STR ameliorates UA (uric acid)-associated renal inflammation, fibrosis, and EMT (epithelial-mesenchymal transition) via MMPS (matrix metalloproteinases), inhibiting NF-κB/NLRP3 activation by the AMPK/SIRT1 pathway and modulating the JAK2-STAT3 and Nrf2 signaling pathways. In summary, the present study provided experimental evidence that STR is an ideal candidate for the treatment of hyperuricemia-mediated renal inflammation. PRACTICAL APPLICATIONS: The higher uric acid results in hyperuricemia and gout. The available options for the treatment of hyperuricemia and gout are the use of allopurinol, and colchicine drugs, etc. These drugs possess several undesirable side effect. The polyphenolic compounds are abundantly present in plants, for example, stevia residue extract (STR) exert a positive effect on human health. From this study results, we can recommend that polyphenolic compounds enrich STR could be applied to develop treatment options for the treatment of hyperuricemia and gout.

Rhodiola crenulate alleviates hypobaric hypoxia-induced brain injury via adjusting NF-κB/NLRP3-mediated inflammation.

BACKGROUND: Rhodiola crenulate (R. crenulate), a famous Tibetan medicine, has been demonstrated to possess superiorly protective effects in high-altitude hypoxic brain injury (HHBI). However, its mechanisms on HHBI are still largely unknown. METHODS: Herein, the protective effects and underlying mechanisms of R. crenulate on HHBI of BABL/c mice were explored through in vivo experiments. The mice model of HHBI was established using an animal hypobaric and hypoxic chamber. R. crenulate extract (RCE) (0.5, 1.0 and 2.0 g/kg) was given by gavage for 7 days. Pathological changes and neuronal viability of mice hippocampus and cortex were evaluated using H&E and Nissl staining, respectively. The brain water content (BWC) in mice was determined by calculating the ratio of dry to wet weight of brain tissue. And serum of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH-Px) and lactate dehydrogenase (LDH) were detected via commercial biochemical kits. Synchronously, the contents of total antioxidant capacity (T-AOC), lactic acid (LA), adenosine triphosphate (ATP), succinate dehydrogenase (SDH), pyruvate kinase (PK), Ca2+-Mg2+-ATPcase, Na+-K+-ATPcase, TNF-α, IL-1ß and IL-6 in brain tissue were quantitative analysis by corresponding ELISA assay. Subsequently, NLRP3, ZO-1, claudin-5, occluding, p-p65, p65, ASC, cleaved-caspase-1, caspase-1 and IL-18 were determined by immunofluorescent and western blot analyses. RESULTS: The results demonstrated that RCE remarkably alleviated pathological damage, BWC, as well enhanced neuronal viability. Furthermore, the oxidative stress injuries were reversely abrogated after RCE treatment, evidenced by the increases of SOD, GSH-Px and T-AOC, while the decreases of MDA and LDH contents. Marvelously, the administration of RCE rectified and balanced the abnormal energy metabolism via elevating the levels of ATP, SDH, PK, Ca2+-Mg2+-ATPcase and Na+-K+-ATPcase, and lowering LA. Simultaneously, the expression of tight junction proteins (ZO-1, claudin-5 and occludin) was enhanced, illustrating RCE treatment might maintain the integrity of blood-brain barrier (BBB). Additionally, RCE treatment confined the contents of IL-6, IL-1ß and TNF-α, and attenuated fluorescent signal of NLRP3 protein. Concurrently, the results of western blot indicated that RCE treatment dramatically restrained p-p65/p65, ASC, NLRP3, cleaved-caspase-1/caspase-1 and IL-18 protein expressions in brain tissues of mice. CONCLUSION: RCE may afford a protectively intervention in HHBI of mice through suppressing the oxidative stress, improving energy metabolism and the integrity of BBB, and subsiding inflammatory responses via the NF-κB/NLRP3 signaling pathway. As a promising agent for the treatment of mice HHBI, the deep-crossing molecular mechanisms of R. crenulate still needs to be further elucidated to identify novel core hub targets.

[Effect of electroacupuncture on pyroptosis of synovial tissues in rats with knee osteoarthritis].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on pyroptosis-related proteins in synovium of knee joint in rats with knee osteoarthritis(KOA), in order to explore its mechanism underlying improvement of KOA. METHODS: Forty male SD rats were randomly divided into control， model, EA and medication groups, with 10 rats in each group. The KOA model was established by injecting 0.2 mL 4% papain solution into the right intra-articular cavity, followed by repeating the injection again on day 4 and 7 after the first injection. After successful modeling, rats of the EA group received EA stimulation of "Neixiyan" (EX-LE4) and "Dubi"(ST35) on the right limb for 15 min, once every day, 6 days per week, for a total of 4 weeks, and those of the medication group received gavage of celecoxib 24 mg/kg, once every day, 6 days per week, for a total of 4 weeks. The severity of dysfunction of the right knee was assessed by using Lequesne's score. Serum interleukin(IL)-1ß and IL-18 contents were detected by ELISA. Histopathological changes of the synovium tissue of the right knee joint were observed to give score (synovial pathological score) after H.E. staining. The expression position and intensity of Nod-like receptor pyrin domain 3 (NLRP3) in syno-vial tissue were observed by immunohistochemistry. The expression levels of NLRP3, apoptosis-associated speck-like protein containing card (ASC), Caspase-1, Gasdermin D(GSDMD), IL-1ß and IL-18 mRNAs and proteins (including GSDMD-N) in the synovial tissue of the right knee joint were detected by real-time fluorescence quantitative PCR and Western blot, separately. RESULTS: Compared with the control group, the model group had a significant increase in the Lequesne's score, synovial pathological score, serum IL-1ß and IL-18 contents, and the expression levels of NLRP3, ASC, Caspase-1, GSDMD, IL-1ß, IL-18 mRNAs and proteins and GSDMD-N protein (P<0.01). Whereas relevant to the model group, both the EA and medication groups had marked lower levels of Lequesne's score and synovial pathological score, serum IL-1ß and IL-18 contents, and expression levels of NLRP3, ASC, Caspase-1, IL-1ß, IL-18 mRNAs and proteins, GSDMD mRNA and GSDMD-N protein (P<0.01, P<0.05). Comparison between two intervention groups showed that the contents of serum IL-1ß and IL-18, and the expression levels of IL-1ß mRNA and protein were significantly higher in the EA group than in the medication group (P<0.05, P<0.01). No significant differences were found between the EA and medication groups in the Lequesne's score, synovial pathological score, NLRP3, ASC, Caspase-1 and IL-18 mRNAs and proteins, as well as GSDMD mRNA (P>0.05). CONCLUSION: EA can alleviate the inflammatory response of synovial tissues of knee joints in KOA rats, which may be related to its function in down-regulating the expression levels of synovial NLRP3, ASC, Caspase-1, IL-1ß and IL-18 mRNAs and proteins, and GSDMD mRNA and GSDMD-N proteins, reducing the occurrence of pyroptosis.

Brassica juncea (L.) Czern. leaves alleviate adjuvant-induced rheumatoid arthritis in rats via modulating the finest disease targets - IL2RA, IL18 and VEGFA.

Brassica juncea (BJ) is a familiar edible crop, which has been used as a dietary ingredient and to prepare anti-inflammatory/anti-arthritic formulations in Ayurveda. But, the scientific validation or confirmation of its therapeutic properties is very limited. This study was performed to determine the efficiency of BJ leaves for the treatment of Rheumatoid arthritis using in vivo and in silico systems. Standard in vitro procedures was followed to study the total phenolic, flavonoid contents and free radical scavenging ability of the extracts of BJ. The effective extract was screened and the presence of bioactive chemicals was studied using HPLC. Further, the possible therapeutic actions of the BJ active principles against the disease targets were studied using PPI networking and docking analysis. IL2RA, IL18 and VEGFA are found to be the potential RA target and the compounds detected from BJ extract have shown great binding efficiency towards the target from molecular docking study. The resulting complexes were then subject to 100 ns molecular dynamics simulation studies with the GROMACS package to analyze the stability of docked protein-ligand complexes and to assess the fluctuation and conformational changes during protein-ligand interactions. To confirm the anti-arthritic activity of BJ, the extract was tested in CFA-induced arthritic Wistar rats. The test groups administered with BJ extract showed retrieval of altered hematological parameters and substantial recovery from inflammation and degeneration of rat hind paw.Communicated by Ramaswamy H. Sarma.

Uso de interferon para el tratamiento de pacientes con COVID-19 / Use of interferon for the treatment of patients with COVID-19

INTRODUCCIÓN: Al momento se tienen propuestas de tratamiento para el COVID-19, en las que se incluye el uso de interferón (Lu 2020; Li and De Clercq 2020). Dentro del espectro de los medicamentos catalogados como interferón (IFN) tenemos IFN alfa o IFN alfa pegilado, IFN alfa 2a, IFN alfa 2b, IFN beta, IFN beta 1a, IFN beta 1b, e IFN gamma (Friedman 2008). De manera anecdótica, ante los brotes de MERS y SARS producidos en años previos, se investigó el uso de IFN como medida terapéutica (Sainz et al. 2004; Scagnolari et al. 2004). Sin embargo, dichos estudios corresponden a estudios in-vitro. En el estudio in vitro de Sainz y col. 2004 (Sainz et al. 2004), se encontró que sinérgicamente IFN beta e IFN gamma inhiben el crecimiento de los cultivos en placa del SARS-CoV, virus que causó el brote del 2003 en China; en el estudio in vitro de Scagnolari y col. 2005 (Scagnolari et al. 2004), se encontró que los IFN presentaban capacidad para inhibir el crecimiento en placa del SARS-CoV (principalmente de IFN beta e IFN gamma). Por otra parte, en una revisión sistemáticade Morra y col 2018 (Morra et al. 2018), donde se estudió el uso de IFN para el tratamiento de MERS-CoV, responsable del brote en Arabia Saudita en el 2012, en sus diferentes presentaciones (incluyendo IFN alfa-2a, IFN alfa-2b, e IFN beta-1a) en combinación con ribavirina, se encontró que no hubo diferencias significativas en desenlaces clínicos cuando se le comparó con otra terapia de soporte. Se observó que se suele utilizar IFN (IFN alfa-2a, IFN alfa-2b, e IFN beta1a) en combinación con ribavirina. En otro estudio tipo revisión sistemática de Momattin y col. 2019 (Momattin, Al-Ali, and Al-Tawfiq 2019), encontraron que no existe consenso para el tratamiento de MERS-CoV pues los estudios fueron heterogéneos y los resultados no concluyentes. En dicho estudio se utilizó IFN 1b en combinación con ribavirina. Debemos resaltar que existe una escasez de evidencia respecto al uso de interferón en COVID19, y la mayoría de la evidencia proviene de estudios in-vitro. GUÍAS DE PRÁCTICA CLÍNICA: La Guía de Medicina Integrativa de China Oriental para el manejo del nuevo coronavirus 2019, provee de información clínica y epidemiológica sobre el COVID 19. Dentro de las alternativas terapéuticas que se encuentran en investigación, menciona al interferón y la evidencia que proviene de estudios de ciencias básicas que encontraron que este medicamento puede frenar la replicación in-vitro del SARS-CoV (Sainz y col. 2004) (Scagnolari y col. 2005). El estudio de Sainz y col. del 2014 encontró que sinérgicamente IFN beta e IFN gamma inhiben el crecimiento de los cultivos en placa del SARS-CoV. El estudio de Scagnolari y col. del 2005, encontró que los IFN presentaban capacidad para inhibir el crecimiento en placa del SARS-CoV (principalmente de IFN beta e IFN gamma). La GPC no presenta recomendaciones respecto al uso de IFN (en ninguna presentación), solo indica que se encuentra en etapa experimental para el uso de pacientes con COVID-19. ESTUDIO SERIE DE CASOS: Lo que se presenta aqui es la experiencia de manejo de pacientes, quienes recibieron múltiples esquemas de tratamiento incluido el IFN. Por tanto, no podemos concluir que los resultados encontrados se deben al uso de IFN. Además, la metodología de una serie de casos no es la ideal para establecer que los pacientes mejoraron por el uso de IFN. Es necesario realizar ensayos clínicos para determinar como influye el IFN en el manejo clínico de pacientes con COVID19. ENSAYOS CLÍNICOS EN CURSO O NO PUBLICADOS REGISTRADOS EN CLINICALTRIALS.GOV: Ensayo clínico no publicado, en fase de reclutamiento de pacientes. NCT04254874 Abidol hydrochloride vs Abidol Hydrochloride combinado con atomización de Interferon PegIFN-α-2b, fase 4, en pacientes con neumonía viral COVID19. Patrocinador Tongji Hospital. A ser realizado en Wuhan, Hubei, China. Fecha estimada de término de estudio: 1ero de julio del 2020. Ensayo clínico no publicado, aún no inicia fase de reclutamiento de pacientes NCT04293887. Uso de interferón alfa-1b en pacientes con infección por COVID19, fase 1, patrocinador Tongji Hospital. Fecha estimada de término de estudio: 30 de junio del 2020. Ensayo clínico no publicado, aún no inicia reclutamiento de pacientes. NCT04275388. Inyección de Xiyanping vs Lopinavir / ritonavir, nebulización de interferon alfa, no aplica fase, en pacientes con neumonía viral COVID19. Patrocinador Jiangxi Qingfeng Pharmaceutical Co. Ltd. Fecha estimada de término de estudio: 14 de diciembre del 2021. Ensayo clínico no publicado, en fase de reclutamiento de pacientes NCT04276688. Lopinavir/ritonavir, ribavirina e interferón beta-1b, fase 2, patrocinador The University of Hong Kong. Fecha estimada de término de estudio: 31 de julio del 2022. EXPERIENCIA RECOGIDA DE HOSPITALES EN EL MUNDO: Debido a que la enfermedad por COVID19 inició en diciembre del 2019, algunos hospitales han recomendado el uso de IFN bajo las siguientes modalidades: España (Sociedad Española de Farmacia Hospitalaria al 18 de marzo 2020): (Sociedad Española de Farmacia Hospitalaria 2020). Se utiliza hasta que caiga la fiebre y no más de 14 días: Estados Unidos (Guía de manejo de COVID19 del Massachusetts Medical Hospital al 17 de marzo 2020): (Massachusetts General Hospital 2020). CONCLUSIONES: A la fecha, 24 de marzo del 2020, aun no se encuentran ensayos clínicos publicados con resultados de eficacia y seguridad respecto al uso del IFN en pacientes con COVID19, infectadas con el virus SARS-CoV-2. Como hecho anecdótico se toma la experiencia de uso de IFN en infecciones causadas por los virus SARS-CoV y MERS-CoV en años previos, los cuales comparten parte importante de su componente genético con el SARS-CoV-2. Encontramos resultados en estudios in-vitro donde el IFN mostró cierta capacidad de inhibir el crecimiento de los virus mencionados. Las recomendaciones de la GPC de Chan y col. (2020), se basa en estudios invitro realizados en el 2004 (Sainz et al. 2004; Scagnolari et al. 2004). La guía de Chan y col. (2020) indica que el uso de IFN en cualquiera de sus presentaciones no se encuentra recomendado para pacientes con infección por COVID-19 y su uso solo es experimental. Por otro lado, la GPC de National Health Commission & Stare Administration of Traditional Chinese Medicine recomienda incluir el uso de IFN-alfa pero no menciona la Encontramos una serie de casos en China (Wan et al. 2020) con 135 pacientes con COVID19 a quienes se le administró IFN más lopinavir/ritonavir, en combinación con antibióticos y corticoides. Alrededor del mes de seguimiento, 11.1% (n=15) de pacientes fueron dados de alta, la tasa de mortalidad a los 28 días de seguimiento fue de 2.5%. Este estudio constituye un primer aporte a la literatura científica sobre el manejo de pacientes con COVID19. Debido a que todos los pacientes del estudio recibieron IFN en combinación con lopinavir/ritonavir, no se puede discriminar si el efecto del IFN es favorable o no para pacientes con COVID19. Es necesario realizar ensayos clínicos para determinar cómo influye el IFN en el manejo clínico de pacientes con COVID19. fuente de donde se obtiene la evidencia para recomendar el uso de IFN. Encontramos que se están llevando a cabo varios ensayos clínicos (EC) respecto al uso de IFN para pacientes con COVID19. Los ensayos clínicos más próximos para finalizar son en julio del 2020. La intervención de interés en la experiencia en hospitales (Sociedad Española de Farmacia Hospitalaria 2020) (Massachusetts General Hospital 2020), por lo general, es dual. Es decir, combina la acción de un tipo de IFN junto a otro antiviral, que por lo general es ribavirina. Otros tipos de antivirales con los cuales se combina al IFN son los esteroides, lopinavir/ritonavir, o micofenolato mofetil. Encontramos guías de sociedades/hospitales internacionales donde actualmente tienen picos muy altos de pacientes con COVID19. Dichas guías incluyen en su algoritmo terapéutico al IFN en diferentes formas (IFN alfa-2b nebulizada e IFN beta-1b inyectable). Sin embargo, no existe ningún ensayo clínico a la fecha que respalde su uso, salvo la experiencia de manejo de pacientes en China. Por ello, son necesarios los resultados de los ensayos clínicos en curso. Por lo expuesto, al momento, no se encuentra que IFN en ninguna de sus presentaciones (IFN alfa o IFN alfa pegilado, IFN alfa 2a, IFN alfa 2b, IFN beta, IFN beta 1a, IFN beta 1b, o IFN gamma), tenga evidencia clínica que respalde una recomendación a favor como una alternativa de tratamiento para pacientes con COVID19. Se necesita de los resultados de los EC que se están realizando para conocer tanto su eficacia como su seguridad en pacientes con COVID19.

Interleukins 18 and 21: biology, mechanisms of action, toxicity, and clinical activity.

Interleukins 18 and 21 have been described, and the effect of each upon immune response and experimental tumors in animals has been the subject of much recent work. Both interleukins have shown antitumor effects in animals, which in some models are striking for their duration, specificity, and ability to protect against rechallenge with the same tumor. These characteristics suggest immunologic involvement in the antitumor response, and several papers suggest involvement of both innate and adaptive immune mechanisms. Recent early phase I clinical trials in human cancer patients have demonstrated evidence of clinical response. This review discusses the biology, preclinical animal tumor model data, and early clinical trial findings.