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OBJECTIVE@#To study the mechanism of the anti-tumor effect of Morinda citrifolia (noni).@*METHODS@#The influences of noni juice on cell proliferation, apoptosis, invasion, migration and the activity of AKT/nuclear factor- κ B (NF- κ B) signaling pathway in A549 human lung cancer cells were detected by MTT, cell counting kit-8, colony formation, Annexin V/PI double labeling, transwell, scratch test and immunoblotting assay, respectively. A549 cells were inoculated into the right axilla of nude mice, followed by noni juice treatment. The body weight of the nude mice was weighed, and the tumor volume and weight were measured. Cell proliferation and expression of apoptosis-related proteins were measured by immunohistochemistry, and the activity of NF- κ B signaling pathway was measured by immunoblotting.@*RESULTS@#The in vitro studies showed that noni juice inhibited the A549 cells proliferation, migration and invasion. Noni juice also promoted cells apoptosis in A549 cells. Immunoblotting assay showed that the phosphorylation level of AKT, p50, and STAT3 proteins was inhibited to different extents after noni juice treatment. The in vivo studies showed that noni juice effectively suppressed tumor formation of A549 cells in nude mice. Noni juice treatment inhibited the expression of Ki67, PCNA, and Bcl-2 protein in the tumor; while promoted the expression of caspase-3 protein. Additionally, we also found that noni juice treatment could restrain the activity of AKT/NF- κ B signaling pathway in the tumor tissue.@*CONCLUSION@#Noni juice inhibited the proliferation of A549 lung cancer cells, induced apoptosis, and inhibited cell invasion and migration via regulating AKT/NF- κ B signaling pathway.
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OBJECTIVE：To study the anti-inflammatory effects of Yuyang capsule on bacterial dermatitis model rats and its effect on TLR/NF-κ B pathway. METHODS：Minimum inhibition concentration （MIC）and minimal bactericidal concentration （MBC）of Yuyang capsule against Staphylococcus aureus were determined by microdilution test. Totally 50 SD rats were randomly divided into model group ，positive control group （amoxicillin and clavulanate potassium ，120 mg/kg as amoxicillin ），Yuyang capsules high-dose ，medium-dose and low-dose groups （according to MIC ），with 10 rats in each group. The model of bacterial dermatitis was established by using the burned skin of rats infected with S. aureus . 24 h after modeling ，administration groups were intragastrically given the corresponding drug ，and model group was intragastrically given the same amount of normal saline ，once a day，for consecutive 7 days. The skin healing rate was calculated on the 1st，3rd，5th and 7th day of administration ，and the scab formation，decrustation and healing were recorded. The contents of IL- 1β，IL-6，IL-10，TNF-α，hydroxyproline（HYP），collagen Ⅰ（Col Ⅰ）and Col Ⅲ in skin tissue were detected by ELISA. Morphology changes of skin tissues were observed by HE staining. The ultrastructure was observed by transmission electron microscope. Protein expression of TLR 2，TLR4 and p-NF-κB p65 were detected by Western blotting assay. RESULTS ：MIC and MBC of Yuyang capsule were 25 mg/mL and 50 mg/mL，respectively. Dose of Yuyang capsules high-dose ，medium-dose and low-dose groups were set at 600，300，150 mg/kg. Compared with model group，scab appeared on the injured skin 3 days after administration in Yuyang capsule high-dose and medium-dose groups ，and decrustation appeared on the injured skin of part mice 5-7 days after administration ；the skin healing rate of the positive control group，Yuyang capsule high-dose and medium-dose groups were all significantly increased at each time point. The contents of IL- 1 β，IL-6，IL-10 and TNF-α，pathological score ，protein expression of TLR 2，TLR4 and p-NF-κB p65 were decreased significantly （P＜0.05 or P＜0.01）；the pathological changes such as inflammatory cell infiltration in skin tissue were improved. The contents of HYP，Col Ⅰ and Col Ⅲ were increased significantly in positive control group and Yuyang capsule high-dose group （P＜0.05 or P＜0.01）. There was no statistical significance in most above indexes in positive control group ，Yuyang capsule high-dose and medium-dose groups （P＞0.05）. CONCLUSIONS ：Yuyang capsule can promote skin healing of bacterial dermatitis model rats and shows certain anti-inflammatory effects ；the mechanism may be related to inhibiting TLR/NF-κB signaling pathway and reducing inflammatory response.
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Objective To study the growth difference and possible mechanism between nasopharyngeal carcinoma (NPC) cell line CNE-2 and its subclone S-18.Methods CNE-2 and S-18 cells were cultured in vitro.6 x 105 cells/mouse were xenografted subcutaneously in the back of nude mice.The volumes of rumors were measured on the 3 rd,7 th,10 th,14 th day after grafting.Mice were sacrificed on the 14 th day and tumors were isolated and weighed.RNA from tumor tissues were extracted and transcriptional levels of HSP27 and NF-K B were detected.Results (1) S-18,instead of CNE-2,grew to form tumor mass 7 days after xenografting subcutaneously;both cell lines formed tumor mass 10 days after xenografting,however,the volumes of S-18 tumors [(223.13 ± 21.32) mm3,10 th day;(420.25 ± 24.52) mm3,14 th day] were significant bigger than CNE-2tumors [(113.70±11.70) mm3,10thday;(279.86±25.78) mm3,14thday];The weights of S-18 umors were significantly higher than CNE-2 tumors on the 14 th day after xenografting;(2) The transcriptional levels of HSP27 and NF-KB in S-18 tumor were significantly higher than in CNE-2 tumor.Conclusion Xenografted S-18 NPC grows faster than Xenografted CNE-2 NPC.HSP27 and NF-κ B are probably involved in the regulation of growth in NPC.
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OBJECTIVE@#To explore effect of high glucose on expression of osteoprotegerin (OPG) and receptor activator of NF- κ B ligand (RANKE) in rat aortic vascular smooth muscle cells.@*METHODS@#SD rats were intraperitoneally injected with streptozotocin, OPG and RANKL expression in rat thoracic aortas were detected by immunohistochemical staining. In cultured vascular smooth muscle cells (VSMCs) (A7r5), qRT-PCR and Western blot analysis were used to examine the mRNA and protein levels of OPG and RANKL.@*RESULTS@#Our results demonstrated that OPG expression was increased in hyperglycemic rat aortic VSMCs, while RANKL expression was decreased. Besides, in vitro experiments high glucose induced OPG expression, but depressed RANKL expression by dose- and time-dependent manner in cultured A7r5.@*CONCLUSIONS@#Our findings suggested that high glucose could promote the expression of OPG, and inhibit the expression of RANKL in VSMCs, which may be partly be the molecular mechanism of diabetic vascular calcification.
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Objective: To explore effect of high glucose on expression of osteoprotegerin (OPG) and receptor activator of NF- κ B ligand (RANKE) in rat aortic vascular smooth muscle cells. Methods: SD rats were intraperitoneally injected with streptozotocin, OPG and RANKL expression in rat thoracic aortas were detected by immunohistochemical staining. In cultured vascular smooth muscle cells (VSMCs) (A7r5), qRT-PCR and Western blot analysis were used to examine the mRNA and protein levels of OPG and RANKL. Results: Our results demonstrated that OPG expression was increased in hyperglycemic rat aortic VSMCs, while RANKL expression was decreased. Besides, in vitro experiments high glucose induced OPG expression, but depressed RANKL expression by dose- and time-dependent manner in cultured A7r5. Conclusions: Our findings suggested that high glucose could promote the expression of OPG, and inhibit the expression of RANKL in VSMCs, which may be partly be the molecular mechanism of diabetic vascular calcification.
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Morphine has been reported to suppress human immune response. We aimed to observe the effects of morphine, fentanyl and tramadol on NF- κ B and IL-2 from both laboratory and clinical perspective. Jurkat cells were incubated with ten times clinically relevant concentrations of morphine,fentanyl and tramadol before being stimulated with PMA. NF- κ B binding activity and IL-2 levels were measured. In the clinical study, 150 consenting patients were randomized into 3 groups according to the analgesics used in them, namely, group morphine (M), group fentanyl (F) and group tramadol (T). IL-2 was measured preoperatively and 1, 3 and 24 h after operation. Consequently, NF-κ B activation was suppressed by morphine and fentanyl but not by tramadol. IL-2 was significantly decreased by morphine and fentanyl but not by tramadol in vitro. In the PCA patients, IL-2 was decreased in group M and increased in group F postoperatively. Whereas in group T, IL-2 was unchanged 1 h after operation but was significantly elevated 3 and 24 h after operation. Our results showed that the inhibition of morphine on IL-2 was most probably related to its suppression on NF-κ B. Fentanyl had different effects on human immune response in vitro and in vivo. Tramadol may have immune enhancing effect.
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· AIM: To study the inhibitory effect of pyrrolidine dithiocarbamate (PDTC) on the inflammatory reaction in an experimental proliferative vitreoretinopathy (PVR)model with laser flare cell meter (LFCM).· METHODS: A total of 20 pigmented rabbits were divided into two groups randomly, with 10 rabbits in each group. After the creation of retinal holes, 0.1mL PDTC was injected intravitreally into the right eyes of Group 1(A1) and the left eyes of Group 1 (A2), and 0.1mL balanced saline solution (BSS) into the right eyes of Group2 (B1). One hour later, 0.1mL BSS into the eyes of A1,and 5000U IL-1 β in 0.1mL BSS was injected intravreally into the eyes of A2 and B1. Clinical evaluation and LFCM examination were performed before retinal injury (PO)and at 4h, 24h, 1, 2 and 4wk after the second injection(P4h, P24h, Plwk, P2wk and P4wk). Histopathologic and immunohistochemical examination were also performed at these time points.· RESULTS: PDTC could inhibit the inflammatory reaction obviously from P24h to P2wk. The eyes of A1 and A2 recovered earlier than those of B1. Although inflammatory reaction in the 3 groups resolved completely by the end of P2wk measured with the slit-lamp microscope,the eyes of the B1 still showed obvious aqueous flare judged by the LFCM compared with those of A1 and A2.Histopathologic and immunohistochemical examination showed that nuclear factor- κ B (NF- κ B) was activated by IL-1 β and the PDTC had inhibitory effect on it without obvious toxicity to retina.· CONCLUSION: Inflammatory reaction involves in the rabbit model of PVR induced by injecting intravitreally IL-1 β and the PDTC can relieve it significantly. The LFCM provides a new, sensitive, objective and noninvasive method to quantify the inflammatory reaction in the PVR model.