Co-Treatment of Phlegm and Stasis Improves Myocardial Inflammation in Rats with Diabetes Mellitus via TLR4/NF-κB/IκB Pathway / 中国实验方剂学杂志

Objective:To observe the intervention of phlegm-stasis co-treatment on the myocardial Toll-like receptor 4 （TLR4）/nuclear factor-<italic>κ</italic>B （NF-<italic>κ</italic>B）/nuclear factor-<italic>κ</italic>B inhibitor （I<italic>κ</italic>B） signaling pathway， and to investigate its mechanism in improving myocardial inflammation in rats with diabetes mellitus （DM）. Method:Forty-five male SD rats of SPF grade were randomly divided into a normal group， a phlegm-resolving （Xiao Xianxiongtang， 4.05 g·kg^-1） group， a stasis-resolving （Xuefu Zhuyutang， 7.02 g·kg^-1） group， a co-treatment （Didang Xianxiong decoction， 8.10 g·kg^-1） group， an alagebrium chloride （3 mg·kg^-1） group， and a model group. Except for normal group， the other rats was induced by a single intraperitoneal injection of 55 mg·kg^-1streptozotocin （STZ） to establish DM model. After adaptive feeding for three weeks， the rats were treated correspondingly by gavage daily for eight weeks. Rats were sampled under anesthesia. Enzyme-linked immunosorbent assay（ELISA） was used to detect the protein expression of TLR4 and tumor necrosis factor-alpha （TNF-<italic>α</italic>） in myocardial tissues. The expression levels of NF-<italic>κ</italic>B p65 and I<italic>κ</italic>B<italic>α</italic> were detected by immunohistochemistry. NF-<italic>κ</italic>B p65， I<italic>κ</italic>B<italic>α</italic>， TNF-<italic>α</italic>， and TLR4 mRNA expression levels were detected by real-time fluorescence-based quantitative polymerase chain reaction（Real-time PCR）. Result:The protein and mRNA levels of TLR4， NF-<italic>κ</italic>B p65， I<italic>κ</italic>B<italic>α</italic>， and TNF-<italic>α </italic>were higher in the model group than those in the normal group （<italic>P</italic><0.01）. TLR4， NF-<italic>κ</italic>B p65， I<italic>κ</italic>B<italic>α</italic>， and TNF-<italic>α</italic> protein and mRNA expression levels were reduced to varying degrees in the groups with drug intervention as compared with those in the model group （<italic>P</italic><0.01）. The inter-group comparison revealed that the co-treatment group showed more manifest reduction in protein and mRNA expression levels of TLR4， NF-<italic>κ</italic>B p65， I<italic>κ</italic>B<italic>α，</italic> and TNF-<italic>α </italic>than the phlegm-resolving group and the stasis-resolving group （<italic>P</italic><0.05<italic>，P</italic><0.01）. Conclusion:The co-treatment of phlegm and stasis can improve myocardial inflammation in DM rats， with superior effect to either the phlegm-resolving method or the stasis-resolving method. The underlying mechanism may be related to the inhibition of TLR4/NF-<italic>κ</italic>B/I<italic>κ</italic>B signaling pathway activation.

The progresses of mechanistic studies of the effect of NF-κB on radiation damage / 中华放射医学与防护杂志

The number of people needed to accept radiotherapy is increasing with the higher incidence of tumor,and the continuous development of radiation therapy technology has extended the longterm survival of patients.Avoiding the occurrence of radiation damage is particularly important.Since NF-κB plays an important role in gene transcription and regulation of radiation damage,this article introduced the structure,activation and function of NF-κB,reviewed the findings of NF-κB alterations in radiation injuries of brain and lung,and sketched the studies of NF-κB inhibitor.

Screening of anti-inflammatory components in Mangiferae Indicae Semen based on bioactivity-integrated UPLC-Q/TOF analysis / 中草药

Objective: To investigate the anti-inflammatory effects of Mangiferae Indicae Semen (MIS) and to screen its bioactive components on nuclear factor-κappa B (NF-κB) inhibition. Methods: The anti-inflammatory effects of MIS were assessed in the swelling of mouse ear induced by dimethylbenzene and the HEK293 inflammatory models induced by TNF-α. UPLC-Q/TOF MS coupled with NF-κB activity luciferase reporter assay system was applied to detect the potential anti-inflammatory components in MIS extract. Results: MIS extract could ameliorate the edema in swelling of mouse ear induced by dimethylbenzene. Ten components were screened to have the potential NF-κB inhibitory effects based on the bioactivity-integrated UPLC-Q/TOF assay system. MIS could contribute to the alleviation of inflammation in the swelling of mouse ear and inhibit the expression of NF-κB. Ten potential active ingredients were found to have anti-inflammatory effects as NF-κB inhibitors, which were mangiferin and gallotannins. The gallotannins included gallic acid, 1-galloyl-β-D-glucopyranosyl-(1→4)-β-D-galactopyranoside, 1-galloyl-β-D-glucopyranosyl- (l→6)-β-D-galactopyranoside, 1,2,3-tri-O-galloyl-β-D-glucose, 1,2,3,4-tetragalloyl-β-D-glucose, 1,2,3,4,6-pentagalloylglucose, and hexagalloylglucose. Conclusion: Mangiferin and gallotannins are verified to be the main bioactive compounds in MIS.

Expression of cerebral Kuppel-like factor 2 in focal cerebral ischemia-reperfusion injury in rats and intervention effect of nuclear factor kappa B inhibitor / 中国脑血管病杂志

Objective To investigate the expression of Kuppel-like factor 2( KLF2 )after focal cerebral ischemia-reperfusion( I/R)injury in rats and the intervention effect of nuclear factor kappa B ( NF-κB)inhibitor. Methods Sixty healthy male SD rats were randomly divided into a sham operation group,an I/R group,and a NF-κB inhibitor group( n=20 in each group). A focal cerebral I/R model was induced by the intraluminal suture method,and NF-κB inhibitor( pyrrolidinedithio carbamate,PDTC)was given to intervene. The observation time points were 6,12,24,and 48 hours after I/R. Reverse transcription-polymerase chain reaction(PCR)and Western blot were used to measure KLF2 mRNA and protein expression in ischemic brain tissue. Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of serum tumor necrosis factorα( TNF-α),and they were compared among groups. Results Compared with the sham operation group,the expression levels of KLF2 mRNA and protein in I/R group in the ischemic brain tissue at each time point were averagely decreased( the relative expression levels of KLF2 mRNA:0. 46 ± 0. 03 vs. 0. 82 ± 0. 04,0. 30 ± 0. 04 vs. 0. 78 ± 0. 05,0. 18 ± 0. 04 vs. 0. 76 ± 0. 02,0. 26 ± 0. 02 vs. 0. 81 ± 0. 04,respectively;the relative expression levels of KLF2 protein:0. 46 ± 0. 04 vs. 0. 80 ± 0. 02,0. 30 ± 0. 02 vs. 0. 79 ± 0. 02,0. 15 ± 0. 02 vs. 0. 77 ± 0. 01,0. 24 ± 0. 01 vs. 0. 79 ± 0. 02,respectively). They reached the lowest values at 24 hours after I/R,while the serum TNF-αlevels were increased. There were significant differences(all P<0. 05). After giving NF-κB inhibitor PDTC,the expression levels of KLF2 mRNA and protein at 6,12,24,and 48 hours after I/R were upregulated differently compared with the I/R group. The relative expression levels of KLF2 mRNA were 0. 61 ± 0. 04,0. 44 ± 0. 03,0. 34 ± 0. 02,and 0. 43 ± 0. 04, respectively. Those of KLF2 protein were 0. 60 ± 0. 02,0. 43 ± 0. 02,0. 33 ± 0. 01,and 0. 44 ± 0. 03, respectively,while the levels of TNF-αwere decreased. There were significant differences(all P<0. 05). There was a negative correlation between the KLF2 mRNA levels and the serum TNF-αlevels at each time point in the I/R group and the PDTC group( r= —0. 728 ,P<0. 05 ). Conclusions The expression levels of KLF2 mRNA in brain tissue are decreased after I/R,and it is negatively correlated with the serum TNF-α levels. It may be involved in the pathological process of I/R by NF-κB pathway mediated inflammatory reaction.

Study on anti-inflammatory components in Rosae Rugosae Flos based on bioactivity-integrated UPLC-Q/TOF method / 中草药

Objective: To investigate the anti-inflammatory effect of Rosae Rugosae Flos (RRF) and screen its bioactive components on nuclear factor-κappa B (NF-κB) inhibition by ultra-performance liquid chromatography coupled with quadrupole/time of flight mass spectrometry (UPLC-Q/TOF MS). Methods: The anti-inflammatory effect of RRF was assessed in Pseudomonas aeruginosa strain-induced acute lung infection mouse model. UPLC-Q/TOF MS coupled with NF-κB activity luciferase reporter assay system was applied to detect the potential anti-inflammatory components in RRF extract. Results: The administration of RRF extract could ameliorate the neutrophil infiltration and diminish the levels of inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interlukin-6 (IL-6), and IL-8 in P. aeruginosa induced mice. Seven components were screened to have potential NF-κB inhibitory effects based on the bioactivity-integrated UPLC-Q/TOF assay system. Conclusion: RRF can contribute to the alleviation of inflammation in mice with acute lung infection induced by P. aeruginosa strain. Seven potentially active ingredients, such as gallic acid, 4-O-galloylquinic acid, methylgallate, centaureidin, strictinin, casuariin, and ellagic acid are found to have anti-inflammatory effects as NF-κB inhibitors.

Effect of NF-κB inhibitor pyrrolidine dithiocarbonate on the proliferation and apoptosis in K562 cells / 中国病理生理杂志

AIM: To investigate the effect of pyrrolidine dithiocarbamate (PDTC), a specific inhibitor of NF-κB on the proliferation and apoptosis of K562 cells and to explore the anti-tumor mechanism of PDTC.METHODS: Trans AM~(TM) NF-κB p65 kit was used to detect the activity of p65 in K562 cells treated by PDTC. The effect of PDTC on the proliferation of K562 cells was measured by WST-1 method. DNA damage was detected by single cell gel electrophoresis (comet assay). The procaspase-3 and activated protein level of caspase-3 were detected by Western blotting.RESULTS: The activity of p65 in K562 cells was inhibited after treated by PDTC (P<0.01). Simultaneously the cell proliferation was significantly inhibited in a dose-and time-dependent manner (P<0.01). The degree of DNA damage in K562 cells treated with PDTC at concentrations of 25 μmol/L, 50 μmol/L or 100 μmol/L was more severe than that in control. The rates of comet cells in the PDTC-treated groups (43.50%, 84.00%, 95.63%) were significantly higher than those in control (9.75%, P<0.01), and it was also dose-dependent. The expression of procaspase-3 and activated caspase-3 protein were detected in the cytoplasm of the K562 cells treated by PDTC by Western blotting.CONCLUSION: NF-κB plays an important role in regulating cell proliferation and apoptosis in K562 cells. PDTC inhibits NF-κB activity and elevates the expression of caspase-3, which is related to increase in cell apoptosis.