Apigenin-Loaded Solid Lipid Nanoparticle Attenuates Diabetic Nephropathy Induced by Streptozotocin Nicotinamide Through Nrf2/HO-1/NF-kB Signalling Pathway

Boschniakine, a bioalkaloid isolated from the Tecoma stans plant, has been reported to be traditionally utilized for the management of diabetes in model systems through multiple biochemical mechanisms. Studies on Boschniakine are sparse, also, taking into consideration that poor absorption, a rapid metabolic rate, and rapid systemic elimination could limit bioalkaloids’ pharmacological efficacy. To recognize such limitations, we propose the use of highly bioavailable Boschniakine-Solid Lipid Nanoparticles (Bos-SLNPs). This study examined the potential molecular mechanisms underlying the anti-diabetic effects of Bos-SLNP. Bos-SLNP (25 and 50 mg/kg) was evaluated for 30 days for its antidiabetic activity in streptozotocin-induced Type 1 and Type 2 diabetic rats. In addition, the RT-PCR technique was used to determine messenger-RNA expression of Interleukin-1, Interleukin-6, and Tumor Necrosis Factor-alpha in renal tissue. haematoxylin and eosin staining and Western blotting were used to find differences in the nuclear factor erythroid 2-related factor 2/heme oxygenase/Nuclear Factor-κB signaling pathway and how its proteins were expressed. Bos-SLNPs treatment resulted in an increase in the expression of nuclear factors erythroid 2-related factor 2 and heme oxygenase-1 and a decrease in the expression of Nuclear factor-B. This was because Bos-SLNPs have a protective mechanism, which comes from their anti-inflammatory and anti-oxidant properties. Bos-SLNPs exhibited a protective effect against diabetic nephropathy in rats induced with streptozocin, possibly via the nuclear factor erythroid 2-related factor pathway 2 / heme oxygenase 1/ nuclear factor B.

BackgroundMany synthesized drugs with clinical severe side effects have been used for diabetic nephropathy (DN) treatment. Therefore, it is urgent and necessary to identify natural and safe agents to remedy DN. Timosaponin B-II (TB-II), a major steroidal saponin constituent in Anemarrhena asphodeloides Bunge, exhibits various activities, including anti-inflammatory and hypoglycemic functions. However, the anti-DN effects and potential mechanism(s) of TB-II have not been previously reported.PurposeTo investigate the effect of TB-II on DN in alloxan-induced diabetic mice.MethodsTB-II was isolated and purified from A. asphodeloides Bunge using macroporous adsorption resin and preparative high-performance liquid chromatography. The effect of TB-II on DN was evaluated in alloxan-induced diabetic mice using an assay kit and immunohistochemical determination in vivo. The expression of mammalian target of rapamycin (mTOR), thioredoxin-interacting protein (TXNIP), and nuclear transcription factor-κB (NF-κB) signaling pathways was also measured using Western blot analysis.ResultsTB-II significantly decreased the blood glucose levels and ameliorated renal histopathological injury in alloxan-induced diabetic mice. In addition, TB-II remarkably decreased the levels of renal function biochemical factors, such as kidney index, blood urea nitrogen, serum creatinine, urinary uric acid, urine creatinine, and urine protein, and it reduced lipid metabolism levels of total cholesterol and triglycerides and the levels of inflammatory cytokines interleukin-6 and tumor necrosis factor-α in alloxan-induced mice. Furthermore, TB-II inhibited the expression of mTOR, TXNIP, and NF-κB.ConclusionThe results revealed that TB-II plays an important role in DN via TXNIP, mTOR, and NF-κB signaling pathways. Overall, TB-II exhibited a prominently ameliorative effect on alloxan-induced DN.

The Chinese medicine Fufang Zhenzhu Tiaozhi capsule protects against renal injury and inflammation in mice with diabetic kidney disease

Aucubin, an iridoid glycoside, possesses beneficial bioactivities in many diseases, but little is known about its neuroprotective effects and mechanisms in brain ischemia and reperfusion (IR) injury. This study evaluated whether aucubin exhibited neuroprotective effects against IR injury in the hippocampal CA1 region through anti-inflammatory activity in gerbils. Aucubin (10 mg/kg) was administered intraperitoneally once a day for one week prior to IR. Neuroprotective effects of aucubin were assessed by neuronal nuclei (NeuN) immunofluorescence and Floro-Jade C (FJC) histofluorescence. Microgliosis and astrogliosis were evaluated using immunohistochemistry with anti-ionized calcium binding adapter protein 1 (Iba1) and glial fibrillary acidic protein (GFAP). Protein levels of proinflammatory cytokines interleukin1 beta (IL1β) and tumor necrosis factor alpha (TNFα) were assayed using enzyme-linked immunosorbent assay and Western blot. Changes in toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling pathway were assessed by measuring levels of TLR4, inhibitor of NF-κB alpha (IκBα), and NF-κB p65 using Western blot. Aucubin treatment protected pyramidal neurons from IR injury. IR-induced microgliosis and astrogliosis were suppressed by aucubin treatment. IR-induced increases in IL1β and TNFα levels were significantly alleviated by the treatment. IR-induced upregulation of TLR4 and downregulation of IκBα were significantly prevented by aucubin treatment, and IR-induced nuclear translocation of NF-κB was reversed by aucubin treatment. Briefly, aucubin exhibited neuroprotective effects against brain IR injury, which might be related to the attenuation of neuroinflammation through inhibiting the TLR-4/NF-κB signaling pathway. These results suggest that aucubin pretreatment may be a potential approach for the protection of brain IR injury.

The present study examined the effects of simvastatin on the proliferation, apoptosis and gene expression levels involved in the nuclear factor-κB (NF-κB) signaling pathway in the human acute promyelocytic leukemia NB4 cell line by methyl thiazolyl tetrazolium assay, flow cytometry and the Human NF-κB Signaling Pathway RT2 Profiler™ PCR Array profiles. The results showed that simvastatin significantly inhibited proliferation and induced apoptosis of the NB4 cells in a time- and dose-dependent manner. Changes were noted in the expression levels of 56 genes involved in the NF-κB signaling pathways in the NB4 cells treated with 15 μm simvastatin at 48 h post-incubation, among which, 47 genes were downregulated and 9 were upregulated. In conclusion, simvastatin potentially inhibits the proliferation and induces the apoptosis of NB4 cells through the regulation of the expression levels of genes involved in the NF-κB signaling pathway.

Lycopene is widely used for nutritional supplementation, but the potential benefits in diabetic nephropathy (DN) remains unknown. This study aimed to highlight the therapeutic prospect of lycopene against streptozotocin (STZ)-induced kidney injury in mice. During the process of the experiments, biochemical kits were employed to determine the diabetes-metabolic parameters in STZ-lesioned mice. Routine pathological and ultrastructural observations were screened for the histological changes of kidney tissue. Moreover, immunohistochemical staining was used to investigate the inflammatory conditions expressed in kidney tissue. Furthermore, intrarenal heme oxygenase 1 (HO-1) mRNA level was assayed via RT-PCR and Western blot analyses. The results showed that lycopene alleviated the lesioned signs of DN mice induced by STZ, accompanied with the increase in body weight, reduced serum concentrations of blood sugar and low-density lipoprotein cholesterol (LDL-C), elevated high-density lipoprotein cholesterol (HDL-C) level, and the decrease in urine protein content. In addition, oxidative defense patterns in the kidneys of DN mice were ameliorated, as shown in augmented bioactivities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and in turn lowered malondialdehyde (MDA) content. The immunohistochemical experiment exhibited that lycopene intake contributed to attenuation of nuclear factor-kappa B (NF-κB) and tumor necrosis factor alpha (TNF-α) expressions in kidney tissue. Moreover, intrarenal HO-1 level was up-regulated in the presence of lycopene. Our findings provide the evidence that lycopene protects kidney cells from STZ-induced lesions via inhibiting NF-κB signal pathway for anti-inflammation and attenuating oxidative stress for anti-dysmetabolism.

Gastric cancer is a common digestive tract malignancy that is mainly treated with surgery combined with perioperative adjuvant chemoradiotherapy and biological targeted therapy. However, the diagnosis rate of early gastric cancer is low and both postoperative recurrence and distant metastasis are thorny problems. Therefore, it is essential to study the pathogenesis of gastric cancer and search for more effective means of treatment. The nuclear factor-κB (NF-κB) signaling pathway has an important role in the occurrence and development of gastric cancer and recent studies have revealed that microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) are able to regulate this pathway through a variety of mechanisms. Understanding these interrelated molecular mechanisms is helpful in guiding improvements in gastric cancer treatment. In the present review, the functional associations between miRNAs, lncRNAs and the NF-κB signaling pathway in the occurrence, development and prognosis of gastric cancer were discussed. It was concluded that miRNAs and lncRNAs have complex relations with the NF-κB signaling pathway in gastric cancer. miRNAs/target genes/NF-κB/target proteins, signaling molecules/NF-κB/miRNAs/target genes, lncRNAs/miRNAs/NF-κB/genes or mRNAs, lncRNAs/target genes/NF-Κb/target proteins, and lncRNAs/NF-κB/target proteins cascades are all important factors in the occurrence and development of gastric cancer.

Chlorogenic acid (CGA) is the primary constituent of Caulis Lonicerae, a Chinese herb used for the treatment of rheumatoid arthritis (RA). The present study aimed to investigate whether CGA was able to inhibit the proliferation of the fibroblast-like synoviocyte cell line (RSC-364), stimulated by interleukin (IL)-6, through inducing apoptosis. Following incubation with IL-6 or IL-6 and CGA, the cellular proliferation of RSC-364 cells was detected by MTT assay. The ratio of apoptosed cells were detected by flow cytometry. Western blot analysis was performed to observe protein expression levels of key molecules involved in the Janus-activated kinase/signal transducer and activator of transcription 3 (JAK/STAT) signaling pathway [phosphorylated (p)-STAT3, JAK1 and gp130] and the nuclear factor κB (NF-κB) signaling pathway [phosphorylated (p)-inhibitor of κB kinase subunit α/β and NF-κB p50). It was revealed that CGA was able to inhibit the inflammatory proliferation of RSC-364 cells mediated by IL-6 through inducing apoptosis. CGA was also able to suppress the expression levels of key molecules in the JAK/STAT and NF-κB signaling pathways, and inhibit the activation of these signaling pathways in the inflammatory response through IL-6-mediated signaling, thereby resulting in the inhibition of the inflammatory proliferation of synoviocytes. The present results indicated that CGA may have potential as a novel therapeutic agent for inhibiting inflammatory hyperplasia of the synovium through inducing synoviocyte apoptosis in patients with RA.

Chronic obstructive pulmonary disease (COPD) is the major leading cause of disease with high-mortality worldwide. Cigarette smoke (CS) is a major factor for COPD. CS causes chronic inflammation and oxidative stress, which contributes to lung dysfunction in COPD. Isoliquiritigenin (ILG), a natural flavonoid derived from the root of liquorice, has been reported to possess antiinflammatory and antioxidant activity. In the present study, we tested the mechanism and protective effects of ILG on CS-induced COPD. Mice were exposed to CS for 2 h twice a day for 4 weeks. ILG was given orally 1 h before CS exposure twice a day for 4 weeks. The bronchial alveolar lavage fluid was collected to test the levels of inflammatory cytokines and the number of inflammatory cells. The lung tissues were obtained to evaluate the pathological changes, lung edema, myeloperoxidase (MPO) activity, malondialdehyde (MDA) level, as well as the expression of the nuclear factor-erythroid 2 (Nrf2) and nuclear factor κB (NF-κB) signaling pathways. The results showed that ILG reduced the infiltration of inflammatory cells and the production of inflammatory cytokines. ILG also reversed CS-induced lung pathological injuries, wet/dry ratio, MPO activity, and MDA level. Further research also showed that ILG dose-dependently up-regulated the expression of Nrf2 and down-regulated the expression of NF-κB signaling pathways induced by CS. In conclusion, ILG protected against CS-induced COPD by inhibiting inflammatory and oxidative stress via the regulation of the Nrf2 and NF-κB signaling pathways.

Objective To investigate the protective effect of paeoniflorin on acute pancreatitis with lung injury and its effect on nuclear factor-κB (NF-κB) signaling pathway. Methods Rats were randomly divided into 3 groups (n=26 each): the sham operation group, severe acute pancreatitis (SAP) group, and the paeoniflorin group. After the model of ASP was established, the serum amylase (AMY), interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) expression levels were detected by enzyme linked immunosorbent assay (ELISA). The NF-κB mRNA expression level was detected by real-time fluorescent quantitative polymerase chain reaction(FQ-PCR), and the wet/dry ratio and pathological changes of lung tissue were measured and compared. Results The expression level of AMY [(4 267±1 459), (2 681±822) vs. (1 482±5 240) U/L] in peripheral blood, MPO [(5.47±1.28), (4.39±0.57) vs. (2.17±0.37) U/g] in the lung, and the wet/dry ratio (5.13±0.62, 4.54±0.34 vs. 4.41±0.21) in the lung tissue in SAP group and the paeoniflorin group were significantly higher than in the sham operation group at 6 h after the operation, and the changes were even more significant in 12 h sub-groups. Pathological score of pancrea tissue (7.44±1.57 vs. 11.30±2.27, P=0.021) and lung tissue (0.83±0.82 vs. 1.47±1.31, P=0.029), aortic blood TNF-α [(83.44±15.28) vs. (120.05±25.88) ng/L, P=0.042], IL-1β [(46.33±10.03) vs. (84.75±17.83) ng/L, P=0.036], IL-6 [(72.77±18.51) vs. (92.81±19.92) ng/L, P=0.046] protein levels and NF-κB mRNA in paeoniflorin group were significantly lower than in SAP group 12 h after the operation (P=0.002). Conclusion Peoniflorin may have the positive protection on lung injury in SAP through inhibition of the NF-κB signaling pathway and thus reducing the release of inflammatory factors such as IL-1β, IL-6 and TNF-α. Key words: Peoniflorin; Severe acute pancreatitis; Lung injury; Inflammation

Objective: To investigate the effects of miR-141-3p on proliferation and migration of gastric cancer cells and nuclear factor-κB (NF-κB) signaling pathway. Methods: Human gastric cancer cell line BGC-823 was cultured, and miR-141-3p mimetic (miR-141-3p mimics) was transfected into BGC-823 cells by lipofection. The miR-141-3p overexpressed BGC was constructed. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the transfection effect. The proliferation of BGC-823 cells was determined by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Transwell assay was used to detect the effect of miR-141-3p on BGC-823 cell migration. The expressions of NF-κB p65, p-IKK-α and p-IKB-α protein in NF-κB signaling pathway were detected by western blot. Results: Compared with the control group and the negative control group, the expression level of miR-141-3p in BGC-823 cells of the miR-141-3p group was (2.39±0.27), which was higher than (1.00±0.09) of the control group and (1.01±0.10) of the negative control group (P<0.05). The number of migrating cells in the miR-141-3p group was (47.64±5.65), which was lower than (106.22±12.14) in the control group and (110.40±12.26) in the negative control group (P<0.05). The expression levels of NF-κB p65, p-IKK-α and p-IKB-α protein in BGC-823 cells were down-regulated (P<0.05). Conclusion: MiR-141-3p can inhibit the proliferation and migration of human gastric cancer BGC-823 cells, which may be related to the inhibition of NF-κB signaling pathway activation.

The flavonoid constituents of Aesculus wilsonii, a source of the Chinese medicinal drug Suo Luo Zi, and their in vitro anti-inflammatory effects were investigated. Fifteen flavonoids, including aeswilflavonosides IA-IC (1: - 3: ) and aeswilflavonosides IIA-IIE (4: - 8: ), along with seven known derivatives were isolated from a seed extract. Their structures were elucidated by extensive spectroscopic methods, acid and alkaline hydrolysis, and calculated electronic circular dichroism spectra. Among them, compounds 3: and 7: possess a 5-[2-(carboxymethyl)-5-oxocyclopent-yl]pent-3-enylate or oleuropeoylate substituent, respectively, which are rarely reported in flavonoids. Compounds 2, 3, 7: , and 12: - 15: were found to inhibit lipopolysaccharide-induced nitric oxide production in RAW 264.7 cell lines. In a mechanistic assay, the flavonoid glycosides 2, 3: , and 7: reduced the expressions of interleukin-6 and tumor necrosis factor-alpha induced by lipopolysaccharide. Further investigations suggest that 2: and 3: downregulated the protein expression of tumor necrosis factor-alpha and interleukin-6 by inhibiting the phosphorylation of p38. Compound 7: was found to reduce the production of inducible nitric oxide synthase, and the secretion of tumor necrosis factor-alpha and interleukin-6 through inhibiting nuclear factor kappa-light-chain-enhancer of activated B signaling pathway. Compounds 2, 3: , and 7: possessed moderate inhibitory activity on the expression of signal transducer and activator of transcription-3. Taken together, the data indicate that the flavonoid glycosides of A.wilsonii seeds exhibit nitric oxide release inhibitory activity through mitogen-activated protein kinase (p38), nuclear factor kappa-light-chain-enhancer of activated B, and signal transducer and activator of transcription-3 cross-talk signaling pathways.

The aim of this article is to identify the relationship between the nuclear factor κB (NF-κB) signal pathway expression and the anastomotic stricture. It particularly focuses on the expression of p65, bcl-2, and cIAP-1 in the anastomotic stricture formation after radical resection of esophageal cancer. A total of 82 esophageal squamous carcinoma patients who had undergone esophagectomy by Ivor Lewis procedure were enrolled in the study. Tissues were collected from the patients who developed the anastomotic stricture after the operation, while para anastomotic tissues served as the control. The protein expression of p65, bcl-2, and cIAP-1 was analyzed by immunohistochemistry and western blot analyses, while the messenger RNA (mRNA) levels of p65, bcl-2, and cIAP-1 were evaluated by reverse transcription polymerase chain reaction (RT-PCR). The results showed that lower expression of p65, bcl-2, and cIAP-1 proteins was observed in the para anastomotic tissue; in the esophageal stricture’s tissue, the expression of these proteins was significantly higher ( P &lt; 0.05). The mRNA levels of P56, bcl-2, and cIAP-1 in the stricture tissue were remarkably increased ( P &lt; 0.05) compared with the para anastomotic tissues, and the mRNA levels in the sample of grade 3 dysphagia were higher ( P &lt; 0.05) than the levels of grade 1 and 2. In the normal esophageal epithelial cell of stricture patients was upregulated compared with that of no stricture patients. We can confirm that the anastomotic stricture has the relationship with the NF-κB signal pathway. The bcl-2 and cIAP-1, located downstream of NF-κB signal pathway, could be a new preventive and therapeutic target for the anastomotic stricture.

Long noncoding RNA urothelial carcinoma-associated 1 (UCA1) has been reported to be a vital mediator in various cancers. But, in terms of gastric cancer (GC), the effects of UCA1 on GC cell proliferation, migration, invasion, and apoptosis remain unclear. This study aimed to uncover the potential regulatory mechanism of UCA1 in GC cells. The expression level of UCA1 was first examined in the five different cell lines of HEK293, CCL-153, HUVEC, SUN-216, and SGC-7901 using a reverse-transcriptase quantitative polymerase chain reaction. Then, the vectors of short hairpin UCA1, the microRNA-182 (miR-182) mimic/inhibitor, and the pEX-tissue inhibitor of metalloproteinases 2 (TIMP2)/small interfering TIMP2 were transfected into SUN-216 and SGC-7901 cells to alter UCA1, miR-182, and TIMP2 expression. To investigate the biological functions, cell viability, migration, invasion, and apoptosis were examined by Cell Counting Kit-8, Transwell, and flow cytometry. The key factors of apoptosis and phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/glycogen synthase kinase 3β (GSK3β) and nuclear factor κB (NF-κB) signal pathways were determined by Western blot analysis. UCA1 was upregulated in SUN-216 and SGC-7901 cells than in the other three cell lines of HEK293, CCL-153, and HUVEC. Knockdown of UCA1 significantly suppressed cell viability, migration, and invasion, and promoted apoptosis by regulating B-cell lymphoma 2, Bax, and cleaved-caspase-3/9 expressions. However, miR-182 overexpression markedly reversed the regulatory effect of UCA1 knockdown on SUN-216 and SGC-7901 cells. TIMP2 was a direct target gene of miR-182, and TIMP2 overexpression exhibited the same effect of UCA1 knockdown on cell viability, migration, invasion, and apoptosis. Besides, miR-182 activated PI3K/AKT/GSK3β and NF-κB signal pathways by regulation of TIMP2. Knockdown of UCA1 exerts an anticancer effect on GC cells by regulating miR-182.

Diabetic nephropathy (DN) is a fatal complication of diabetes and the main cause of end-stage renal disease. Due to the suboptimal effects of current treatments, there is an urgent need to develop new therapeutic strategies for DN. Trametenolic acid (TA), a lanostane-type tetracyclic triterpenoid, is one of the main active ingredients extracted from the natural product Inonotus obliquus. Our study was aimed at clarifying the potential protective effects of TA on DN and its underlying mechanism. In this research, C57BLKS/db (db/db) mice were used as the spontaneous DN model, and TA (10 mg/kg/d) was intraperitoneally injected for 4 consecutive weeks. Ratio of right kidney weight/body weight was calculated, and the contents of serum creatinine (Scr), blood urea nitrogen (BUN), and urine albumin were detected. The activities of superoxide dismutase (SOD) and catalase (CAT) and the contents of reductive glutathione (GSH) and malondialdehyde (MDA) were measured. The histopathological changes of renal tissues were observed by hematoxylin and eosin (HE), periodic acid-Schiff (PAS), and Masson staining. The protein expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), NAD(P)H:quinone oxidoreductase-1 (NQO-1), nuclear factor kappa B (NF-κB), proinflammation cytokine tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), Nephrin, and Podocin were detected by western blot assay. Immunohistochemistry was utilized to detect expressions of collagen III (COL-III) and fibronectin (FN). Our results showed that TA administration significantly reduced the ratio of right kidney weight/body weight, BUN, Scr, and urine albumin levels and alleviated the histopathological changes of DN mice. Moreover, TA administration remarkably increased GSH content and SOD and CAT activities and decreased MDA content. Western blot assay demonstrated that TA activated Nrf2 signaling and increased the expression of downstream antioxidant enzymes HO-1 and NQO-1. Further studies illustrated that NF-κB signaling was inhibited, and downstream proinflammation cytokine expressions of TNF-α, IL-6, and IL-1β were also downregulated. In addition, we also found that TA administration significantly increased the expression of nephrin and podocin proteins and reduced the protein expression of COL-III and FN. These findings suggested that TA exhibited a renoprotective effect by ameliorating oxidative stress and inflammation via Nrf2/HO-1 and NF-κB signaling pathways.