Protective effects of electroacupuncture on acetylsalicylic acid-induced acute gastritis in rats

AIM: To investigate the protective effects of electroacupuncture (EA) pretreatment on acetylsalicylic acid (ASA)-induced ulceration in rats. METHODS: We randomly divided 72 rats into three groups including control (administered with distilled water), ASA group (administered 100 mg/kg ASA) and EA group (administered EA + 100 mg/kg ASA). Each rat was fasted for 18 to 24 h before experimentation, and lesion scores, gastric acidity, cyclooxygenase (COX)-1 and -2 mRNA levels, and total nitric oxide (NO) concentration were measured. RESULTS: The lesion scores of the EA group were significantly lower than those of the ASA group. Gastric acidity of the ASA and EA groups was reduced compared to the control group. COX-1 and -2 mRNA levels were significantly increased in the EA group as compared to the control and ASA groups, and NO levels were also significantly increased in the EA group as compared to the ASA group. CONCLUSION: These results suggest that EA-mediated protection against ASA-induced ulceration in rats may occur via gastric defense components.

Inhibitory effect of Patrinia scabiosaefolia on acute pancreatitis

瞄准：在缩胆囊素(CCK ) 上调查 Patrinia scabiosaefolia (PS ) 的效果在老鼠的导致 octapeptide 的尖锐胰腺炎(AP ) 。方法：称 240-260 g 的 Wistar 老鼠被划分成三个组：(1 ) 正常的盐对待的组；(2 ) 在哪个 PS 口头上地被管理，有在 100 mg/kg 组的 PS 的处理在 1， 3 和 5 h，和这个全部以后由 75 microg/kg CCK octapeptide 的下的管理列在后面三次过程为 5 d 被重复；(3 ) 与盐的组，协议与在处理一样在是一起的处理与 PS 组织。我们决定了胰腺的重量 / 身体重量比率，胰腺的 HSP60 的层次， HSP72 和支持 inflammatory cytokines 的分泌物。重复 CCK octapeptide 处理导致了试验性地导致的胰腺炎的典型实验室调查结果。结果：PS 在 CCK 导致 octapeptide 的 AP 减少了胰腺的重量 / 身体重量比率，浆液淀粉酶和脂肪分解酵素的层次，和支持 inflammatory cytokines 的禁止的表情。而且， PS 预告的处理增加了 HSP60 和 HSP72 的胰腺的层次。结论：有 PS 的预告的处理在 CCK 导致 octapeptide 的 AP 上有反煽动性的效果。

Potentiating activity of luteolin on membrane permeabilizing agent and ATPase inhibitor against methicillin-resistant Staphylococcus aureus

Objective:To investigate the mechanism of antibacterial activity of luteoiin(LUT) against methicillin-resistant Staphylococcus aureus(MRSA).Methods:The mechanism of anti-MRSA activity of LUT was analyzed by the viability assay in membrane permeabilizing agent ATPase inhibitors,and peptidoglycan(PGN) derived from Staphylococcus aureus(S.aureus).Also,transmission electron microscopy was used to monitor survival characteristics and changes in S.aureus morphology.Results:Compared to the LUT alone,the optical density of suspensions treated with the combination of 125 μg/mL Tris and 230 μg/mL DCCD were reduced to 60%and 46%,respectively.PGN(15.6 μg/mL) gradually impeded the activity of LUT,and PGN(62.5 μg/mL) completely blocked the activity of LUT on S.aureus.Conclusions:Increased susceptibility to LUT with me Tris and DCCD combinations is evident in all tested MRSA isolates.The results indicate LUT synergy in increasing cytoplasmic membrane permeability and inhibiting ATPase.S.aureus PGN directly blocks the antibacterial activity of LUT,suggesting the direct binding of LUT with PGN.These findings may be validated for the development of antibacterial agent for low MRSA resistance.

Effect of biologically active fraction of Nardostachys jatamansi on cerulein-induced acute pancreatitis

AIM:To determine if the fraction of Nardostachys jatamansi(NJ) has the potential to ameliorate the severity of acute pancreatitis(AP).METHODS:Mice were administered the biologically active fraction of NJ,i.e.,the 4th fraction(NJ4),intraperitoneally,and then injected with the stable cholecystokinin analogue cerulein hourly for 6 h.Six hours after the last cerulein injection,the pancreas,lung,and blood were harvested for morphological examination,measurement of cytokine expression,and examination of neutrophil infiltration.RESULTS:NJ4 administration attenuated the severity of AP and lung injury associated with AP.It also reduced cytokine production and neutrophil infiltration and resulted in the in vivo up-regulation of heme oxygenase-1(HO-1).Furthermore,NJ4 and its biologically active fraction,NJ4-2 inhibited the cerulein-induced death of acinar cells by inducing HO-1 in isolated pancreatic acinar cells.CONCLUSION:These results suggest that NJ4 may be a candidate fraction offering protection in AP and NJ4 might ameliorate the severity of pancreatitis by inducing HO-1 expression.

Antimicrobial activity and synergism of Sami-Hyanglyun-Hwan with ciprofloxacin against methicillin-resistant Staphylococcus aureus

Objective: To investigate the antibacterial activity of SHHextracted with either water or ethanol against methicillin-resistant Staphylococcus aureus(MRSA) and combinatory antimicrobial effect with ciprofloxacin(CIP) by time kill assay and checkerboard dilution test. Methods: The antibacterial activity determined by broth dilution method indicated that the antibacterial activity of Sami-Hyanglyun-Hwan(SHH) water extract(SHHW) and SHH ethanol extract(SHHE) ranged from 250 to 2000 μg/m L and 125 to 1000 μg/m L against MRSA, respectively. Results: In the checkerboard method, the combinations of SHHE with CIP had a partial synergistic or synergistic effect against MRSA. The time-kill curves showed that a combined SHHE and CIP treatment reduced the bacterial counts dramatically after 24 h. Conclusions: The present study demonstrates the therapeutic ability of SHHE against MRSA infections.

Effects of Gastrodiae rhizoma on proliferation and differentiation of human embryonic neural stem cells

Objective:To investigate the effects of Gastrodiae rhizoma,a dried root of Gastrodia elata Blume,on proliferation and differentiation of human NSCs derived from embryonic stem cells.Methods:A 70%ethanol extract of Gastrodiae rhizoma(EEGR) was estimated with4-hydroxybenzyl alcohol as a representative constituent by HPLC.Results:MTT assay showed that the treatment with EEGR increased the viability of NSCs in growth media.Compared to contro1,EEGR increased the number of dendrites and denritic spines extended from a differentiated NSC.Whereas EEGR decreased the mRNA expression of Nestin,it increased that of Tuj1 and MAP2 in NSCs grown in differentiation media.Immunocytochemical analysis using confocal microscopy also revealed the increased expression of MAP2 in dendrites of EEGR-treated NSCs.Furthermore,EEGR decreased mRNA expression of Sox2 in NSCs grown even in growth media.Conclusions:In conclusion,our study demonstrates for the first time that EEGR induced proliferation and neuronal differentiation of NSCs,suggesting its potential benefits on NSC-based therapies and neuroregeneration in various neurodegenerative diseases and brain Injuries.

The inhibition effect of Chlorpromazine against the β-lactam resistance of MRSA

Objective:To investigate the gene related to β-lactam resistance and to confirm the mechanism about a synergy effect between CPZ and β-lactam antibiotics.Methods:To measure antibacterial activity,we performed a minimum inhibitory concentration(MIC) and synergy test.Transmission electron microscopy(TEM) was used in morphological analysis.To analyze gene expression,we conducted reverse transcriptase polymerase chain reaction(PCR).Results:We confirmed a synergy effect between CPZ and β-lactam antibiotics.Furthermore,we observed that CPZ affect the cell envelope of MRS A by using TEM.At the gene level,CPZ reduced the expression of resistance genes.Conclusions:Through this result,we hypothesize that a decrease of resistance factor expressions was caused by CPZ because it disrupts the activity of a sensor protein located in the cell membrane.

Cortex cinnamomi extract prevents streptozotocin- and cytokine-induced β-cell damage by inhibiting NF-κB

AIM: To clarify the mechanism underlying the anti-diabetic activities of cortex cinnamomi extract (CCE). METHODS: To induce in vivo diabetes, mice were injected with streptozotocin (STZ) via a tail vein (100 mg STZ/kg body weight). To determine the effects of CCE, mice were administered CCE twice daily for 7 d by oral gavage starting 1 wk before the STZ injection. Blood glucose and plasma insulin concentration were measured as an index of diabetes. Also, to induce cytotoxicity of RINm5F cells, we treated with cytokines (IL-1β(2.0 ng/mL) and IFN-γ(100 U/mL)). Cell viability and nitric oxide production were measured colorimetrically. Inducible nitric oxide synthase (iNOS) mRIMA and protein expression were determined by RT-PCR and Western blotting, respectively. The activation of NF-κB was assayed by using gel mobility shift assays of nuclear extracts. RESULTS: Treatment of mice with STZ resulted in hyperglycemia and hypoinsulinemia, which was further evidenced by immunohistochemical staining of islets. However, the diabetogenic effects of STZ were completely prevented when mice were pretreated with CCE. The inhibitory effect of CCE on STZ-induced hyperglycemia was mediated through the suppression of iNOS expression. In rat insulinoma RINm5F cells, CCE completely protected against interleukin-1βand interferon-γ-mediated cytotoxicity. Moreover, RINm5F cells incubated with CCE showed significant reductions in interleukin-lp and interferon-γ-induced nitric oxide production and in iNOS mRNA and protein expression, and these findings correlated well with in vivo observations. CONCLUSION: The molecular mechanism by which CCE inhibits iNOS gene expression appears to involve the inhibition of NF-κB activation. These results reveal the possible therapeutic value of CCE for the prevention of diabetes mellitus progression.

Scolopendra subspinipes mutilans protected the ceruleininduced acute pancreatitis by inhibiting high-mobility group box protein-1

AIM:To evaluate the inhibitory effects of Scolopendra subspinipes mutilans(SSM) on cerulein-induced acute pancreatitis(AP) in a mouse model.METHODS:SSM water extract(0.1,0.5,or 1 g/kg) was administrated intraperitoneally 1 h prior to the first injection of cerulein.Once AP developed,the stable cholecystokinin analogue,cerulein was injected hourly,over a 6 h period.Blood samples were taken 6 h later to determine serum amylase,lipase,and cytokine levels.The pancreas and lungs were rapidly removed for morphological examination,myeloperoxidase assay,and real-time reverse transcription polymerase chain reaction.To specify the role of SSM in pancreatitis,the pancreatic acinar cells were isolated using collagenase method.Then the cells were pre-treated with SSM,then stimulated with cerulein.The cell viability,cytokine productions and high-mobility group box protein-1(HMGB-1) were measured.Furthermore,the regulating mechanisms of SSM action were evaluated.RESULTS:The administration of SSM significantly attenuated the severity of pancreatitis and pancreatitis associated lung injury,as was shown by the reduction in pancreatic edema,neutrophil infiltration,vacuolization and necrosis.SSM treatment also reduced pancreatic weight/body weight ratio,serum amylase,lipase and cytokine levels,and mRNA expression of multiple inflammatory mediators such as tumor necrosis factor-α and interleukin-1β.In addition,treatment with SSM inhibited HMGB-1 expression in the pancreas during AP.In accordance with in vivo data,SSM inhibited the cerulein-induced acinar cell death,cytokine,and HMGB-1 release.SSM also inhibited the activation of c-Jun NH2-terminal kinase,p38 and nuclear factor(NF)-κB.CONCLUSION:These results suggest that SSM plays a protective role during the development of AP and pancreatitis associated lung injury via deactivating c-Jun NH2-terminal kinase,p38 and NF-κB.

Selective cyclooxygenase-2 inhibitor ameliorates cholecystokinin-octapeptide-induced acute pancreatitis in rats

AIM: To investigate the effect of selective Cycloo- xygenase-2 (COX-2) inhibitor 4-[5-(4-Chloro-phenyl)-3- (trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide (SC-236), on the cholecystokinin (CCK)-octapeptide- induced acute pancreatitis (AP) in rats. METHODS: Wistar rat weighing 240 g to 260 g were divided into three groups. (1) Normal DMSO treated group, (2) SC-236 at 4 mg/kg treated group; SC-236 systemically administered via the intravenous (i.v.) catheter, followed by 75 μg/kg CCK octapeptide subcutaneously three times, after 1, 3 and 5 h. This whole procedure was repeated for 5 d. (3) Dimethyl sulfoxide (DMSO) treated group: an identical protocol was used in this group as in the SC-236 cohort (see 2. above). Repeated CCK octapeptide treatment resulted in a typical experimentally induced pancreatitis in the Wistar rats. RESULTS: SC-236 improved the severity of CCK- octapeptide-induced AP as measured by laboratory criteria [the pancreatic weight/body weight (p.w/ b.w) ratio, the level of serum amylase and lipase]. The SC-236 treated group showed minimal histologic evidence of pancreatitis and a significant reduction inmyeloperoxidase activity. SC-236 also increased heat shock protein (HSP)-60 and HSP72 compared with the DMSO-treated group in the CCK-octapeptide-induced AP and also reduced the pancreatic levels of COX-2. Furthermore, SC-236 reduced proinflammatory cytokine synthesis and inhibited NF-κB activation compared with the DMSO-treated group in the CCK-octapeptide-induced AP. CONCLUSION: Our results suggested that COX-2 plays pivotal role in the development of AP and COX-2 inhibitors may play a beneficial role in preventing AP.

Nardostachys jatamansi extract protects against cytokine-induced β-cell damage and streptozotocin-induced diabetes

AIM: To investigate the anti-diabetogenic mechanism of Nardostachys jatamansi extract (NJE). METHODS: Mice were injected with streptozotocin viaa tail vein to induce diabetes. Rat insulinoma RINm5F cells and isolated rat islets were treated with interleukin1β and interferon-γ to induce cytotoxicity. RESULTS: Treatment of mice with streptozotocin resulted in hyperglycemia and hypoinsulinemia, which was conf irmed by immunohistochemical staining of the islets. The diabetogenic effects of streptozotocin were completely abolished when mice were pretreated with NJE. Inhibition of streptozotocin-induced hyperglycemia by NJE was mediated by suppression of nuclear factor (NF)-κB activation. In addition, NJE protected against cytokine-mediated cytotoxicity. Incubation of RINm5F cells and islets with NJE resulted in a signif icant reduction in cytokine-induced NF-κB activation and downstream events, inducible nitric oxide synthase expression and nitric oxide production. The protective effect of NJE was further demonstrated by the normal insulin secretion of cytokine-treated islets in response to glucose. CONCLUSION: NJE provided resistance to pancreatic β-cell damage from cytokine or streptozotocin treatment. The β-cell protective effect of NJE is mediated by suppressing NF-κB activation.

Gardenia jasminoides protects against cerulein-induced acute pancreatitis

AIM: To investigate the effect of Gardenia jasminoides (GJ) on cerulein-induced acute pancreatitis (AP) in mice.METHODS: C57BL/6 mice weighing 18-20 g were divided into three groups.(1) Normal saline-treated group,(2) treatment with GJ at a dose of 0.1 g/kg,(3) treatment with GJ at a dose of 1 g/kg.GJ was administered orally (n = 6 per group) for 1 wk.Three hours later,the mice were given an intraperitoneal injection of cerulein (50 μg/kg),a stable cholecystokinin (CCK) analogue,every hour for a total of 6 h as described previously.The mice were sacrificed at 6 h after completion of cerulein injections.Blood samples were obtained to determine serum amylase,lipase and cytokine levels.The pancreas was rapidly removed for morphologic examination and scoring.A portion of pancreas was stored at -70℃ and prepared for the measurement of tissue myeloperoxidase (MPO) activity,an indicator of neutrophil sequestration,and for reverse-transcriptase PCR (RT-PCR) and real-time PCR measurements.RESULTS: Treatment with GJ decreased significantly the severity of pancreatitis and pancreatitis-associated lung injury.Treatment with GJ attenuated the severity of AP compared with saline-treated mice,as shown by reduction in pancreatic edema,neutrophil infiltration,serum amylase and lipase levels,serum cytokine levels,and mRNA expression of multiple inflammatory mediators.CONCLUSION: These results suggest that GJ attenuated the severity of AP as well as pancreatitis-associated lung injury.

Involvement of heme oxygenase-1 induction in anti-vascular inflammation effects of Xanthoceras sorbifolia in human umbilical vein endothelial cells

OBJECTIVE: To define the effects of Xanthoceras sorbifolia(EXS) on vascular inflammation and the mechanisms in endothelial cells.METHODS: Vascular protective effects of an ethanol extract of seeds from EXS(1-50 μg/mL) against tumor necrosis factor-α(TNF-α)-induced vascularinflammation were examined in human umbilical vein endothelial cells(HUVECs).RESULTS: EXS significantly decreased TNF-α-induced expression of cell adhesion molecules, such as intracellular adhesion molecule-1, vascular cell adhesion molecule-1, and endothelial cell selectin,in a dose-dependent manner. Pre-treatment with EXS significantly inhibited translocation and transcriptional activity of nuclear factor-κB(NF-κB) increased by TNF-α. EXS also significantly inhibited formation of intracellular reactive oxygen species(ROS). Moreover, the vascular protective effects of EXS were linked to up-regulation of heme oxygenase-1(HO-1) and nuclear factor E2-related factor-2(Nrf-2) expression. EXS-induced HO-1 expression was significantly decreased in SnPP(HO-1 inhibitor)-and HO-1 siRNA-treated cells, whereas an increase was found in cobalt protoporphyrin IX(CoPP)(HO-1 inducer)-treated cells. In addition, pretreatment with EXS increased HO-1 and Nrf-2 expression under TNF-α stimulation with or without N-acetyl-L-cysteine. Furthermore, the inhibitory effects of EXS on TNF-α-induced vascular inflammation were partially reversed in SnPP-and of HO-1siRNA-treated cells but increased by CoPP.CONCLUSION: These results suggest that EXS may have important implications for prevention of vascular complications associated with vascular inflammation by inhibition of the NF-κB/ROS pathway and activation of the Nrf-2/HO-1 pathway.

Effects of Yiqi Huoxue Recipe （益气活血方） and Coxsackie Virus B Type 3 on the Expression of Ribosomal Protein S20 in Rat Cardiac Myocytes

Objective： To study the effects of Yiqi Huoxue Recipe （益气活血方） and Coxsackie B virus type 3 （CVB3） on the expression of ribosomal protein S20 in rat cardiac myocytes, to explore the pathogenesis of myocarditis induced by CVB3 and the mechanism of Yiqi Huoxue Recipe on gene level, and to further investigate whether Yiqi Huoxue Recipe is an effective prescription for CVB3 myocarditis. Methods： A modified suppression subtractive hybridization （SSH） was used to isolate differentially expressed genes between the CVB3 infection group and the treatment group with Yiqi Huoxue Recipe. The results were further verified by fluorescence RT-PCR. Results： The results of SSH showed that the gene expression of ribosomal protein S20 in the treatment group was higher than that in the CVB3 infection group （P〈0.05）, which agreed with the results of fluorescent RT-PCR. Conclusion： Down-regulation of ribosomal protein S20 mRNA expression might be one of the mechanisms in CVB3 myocarditis, and Yiqi Huoxue Recipe could achieve the treatment of viral myocarditis by regulating the expression of ribosomal protein S20.

Neuroprotective effects of Suhexiang Wan on the in vitro and in vivo models of Parkinson’s disease

OBJECTIVE:To examine the role of KSOP1009(a modified formulation of Suhexiang Wan essential oil)in an animal model of Parkinson's disease(PD)induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydro-pyridine(MPTP)injection.METHODS:Cell toxicity,apoptosis,and reactive oxygen species(ROS)levels were analyzed in the human neuroblastoma cell line SH-SY5 Y.After that,changes in animal behavior and tyrosine hydroxylase(TH)protein levels in the substantia nigra(SN)of MPTP-injected mice were examined.Three different doses of KSOP1009(30,100,and 300 mg/kg,n=8 for each group)were administered daily for 7 d before MPTP injection and 14 d after MPTP injection,totaling 21 d.RESULTS:MPP+,the active metabolite of MPTP,decreased the viability of SH-SY5 Y cells,whereas KSOP1009 alleviated MPP+-induced cytotoxicity.KSOP1009(10 and 50 mg/m L)reduced MPP+-induced ROS generation compared with the control group.Treatment with 1 m M MPP+increased the percentage of depolarized/live cells,whereas KSOP1009 intake at a dose of 10 mg/m L decreased the percentage of these cells.The mean latency to fall in the rotarod test was reduced in mice treated with MPTP compared with the control group.However,mice receiving three different doses of KSOP1009 performed better than MPTP-treated animals.MPTP-treated mice were more hesitant and took longer to traverse the balance beam than the control animals.In contrast,KSOP1009-treated mice performed significantly better than MPTPtreated mice.Furthermore,the KSOP1009-treated groups had a significantly higher number of TH-positive neurons in the lesioned SN and significantly higher expression of TH in the striatum than the MPTP-treated group.MPTP treatment strongly induced Jun-N-terminal kinase(JNK)activation,whereas KSOP1009 suppressed MPTP-induced JNK activation.In addition,KSOP1009 intake reversed the decrease in the phosphorylation levels of c AMP-response element-binding protein in the brain of MPTP-treated mice.KSOP1009 also restored the decrease in dopaminergic neurons and dopamine levels in the brain of MPTP-treated mice.CONCLUSION:KSOP1009 protected mice against MPTP-induced toxicity by decreasing ROS formation and restoring mitochondrial function.

Beneficial Effect of Berberis amurensis Rupr. on Penile Erection

Objective： To investigate whether the methanol extract of Berberis amurensis Rupr. （BAR） augments penile erection using in vitro and in vivo experiments. Methods： The ex vivo study used corpus cavemosum strips prepared from adult male New Zealand White rabbits. In in vivo studies for intracavernous pressure （ICP）, blood pressure, mean arterial pressure （MAP）, and increase of peak ICP were continuously monitored during electrical stimulation of Sprague-Dawley rats. Results： Preconstricted with phenylephrine （PE） in isolated endothelium- intact rabbit corus cavernosum, BAR relaxed penile smooth muscle in a dose-dependent manner, which was inhibited by pretreatment with NG-nitro-L-arginine methyl ester （L-NAME）, a nitric oxide synthase inhibitor, and 1H-[1,2,4]-oxadiazole-[4,3-α]-quinoxalin-l-one, a soluble guanylyl cclase inhibitor. BAR significantly relaxed penile smooth muscles dose-dependently in ex vivo, and this was inhibited by pretreatment with L-NAME 1H-[1,2,4]- oxadiazole-[4,3-α ]-quinoxalin-l-one. BAR-induced relaxation was significantly attenuated by pretreatment with tetraethylammonium （TEA, P〈0.01）, a nonselective K＋ channel blocker, 4-aminopyridine （4-AP, P〈0.01）, a voltage-dependent K＋ channel blocker, and charybdotoxin （P〈0.01）, a large and intermediate conductance Ca2. sensitive-K＋ channel blocker, respectively. BAR induced an increase in peak ICP, ICP/MAP ratio and area under the curve dose dependently. Conclusion： BAR augments penile erection via the nitric oxide/cyclic guanosine monophosphate system and Ca2. sensitive-K＋ （BK＋ and IK＋） channels in the corpus cavemosum.

Electroacupuncture on PC6 Prevents Opioid-Induced Nausea and Vomiting after Laparoscopic Surgery

Objective： To investigate the treatment time dependence of electroacupuncture （EA） on Neiguan （PC6） for preventing postoperative nausea and vomiting （PONV）. Methods： One hundred and seventy-eight patients, who had received intravenous patient-controlled analgesia （PCA） with Fentanyl, were assigned randomly to three groups using random numbers： a pre-operative EA group （PrEA）, a post-operative EA group （PoEA）, and a non-acupuncture control group （NC）. An anesthetist evaluated the incidence and severity of nausea and vomiting for 48 h after surgery blindly. The main outcomes were severity and freguency of PONV, which were measured with a self-reported questionnaire and a confirmation from the anesthetist. The data were analyzed with ANOVA and Z-test. Results： The incidence of nausea and vomiting was significantly lower in the PrEA group than the NC group during 48 h after surgery （P〈0.01, P〈0.05）. The incidence of vomiting was also significantly lower in the PrEA group than the PoEA group （P〈0.05）. The PoEA subjects evidenced no significant differences compared with the NC subjects in terms of the incidence of nausea and vomiting （P〈0.05）. The severity of nausea was significantly lower in the PrEA group than in the NC and PoEA groups （P〈0.05）. Conclusions： EA on PC6 is effective in the prevention of PONV, and pre-operative acupuncture is more effective than post-operative acupuncture.

Hwangryunhaedoktang exerts anti-inflammation on LPS-induced NO production by suppressing MAPK and NF-κB activation in RAW264.7 macrophages

OBJECTIVE： This study aimed to evaluate whether Hwangryunhaedoktang （HHT）, a herbal compound, has an inhibitory effect on lipopolysaccharide （LPS）-induced inflammation in RAW264.7 macrophages. METHODS： The effects of HHT were evaluated by confirming nitric oxide （NO） production and expression of inducible NO synthase （iNOS） and mitogen-activated protein kinases （MAPKs） in LPS-stimulated RAW264.7 macrophages via the Griess assay, Western blotting, and real-time reverse transcription quantitative polymerase chain reaction. Western blot analyses and luciferase assays were used to evaluate whether HHT has an effect on the phosphorylation and translocation of nuclear factor-κB （NF-κB）. The secretion and expression of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） were determined via enzyme-linked immunosorbent assay and Western blot analyses. RESULTS： HHT suppressed LPS-induced NO production and expression of iNOS in a dose-dependent manner. Additionally, MAPKs activation was also attenuated via inhibition of phosphorylation of extracellular signal-regulated kinases 1/2, c-Jun N-terminal kinase and p38 which were related to inflammatory pathway. Furthermore, HHT also effectively attenuated NF-κB activation and its translocation to the nucleus, a process that is closely linked to inflammation. LPS normally induced the expression of inflammatory cytokines such as TNF-α and IL-6, but the secretion and expression of TNF-α and IL-6 were significantly attenuated by pretreating the cells with HHT. CONCLUSION： HHT suppressed LPS-induced NO production by blocking the activation of NF-κB and MAPK signaling pathways in RAW264.7 macrophages. Furthermore, HHT may have an anti-inflammatory effect by suppressing the LPS-induced secretion of TNF-α and IL-6. Therefore, the traditional herbal formula HHT might be a useful potential therapeutic agent for inflammation.

Sinkihwan-gamibang ameliorates puromycin amino nucleoside-induced nephrotic syndrome

Nephrotic syndrome(NS)is a kidney disease characterized by hypertriglyceridemia,massive proteinuria,hypo-albuminemia and peripheral edema.Sinkihwan-gamibang(SKHGMB)was recorded in a traditional Chinese medical book named“Bangyakhappyeon(方藥合編)”and its three prescriptions Sinkihwan,Geumgwe-sinkihwan,and Jesaeng-sinkihwan belong to Gamibang.This study confirmed the effect of SKHGMB on renal dysfunction in an NS model induced by puromycin aminonucleoside(PAN).The experimental NS model was induced in male Sprague Dawley(SD)rats through injection of PAN(50 mg·kg^(-1))via the femoral vein.SKHGMB not only reduced the size of the kidneys increased due to PAN-induced NS,but also decreased proteinuria and ascites.In addition,SKHGMB significantly ameliorated creatinine clearance,creatinine,and blood urea nitrogen.SKHGMB relieved glomeruli dilation and tubules fibrosis in the glomeruli of the NS model.SKHGMB inhibited the protein and mRNA levels of the NLRP3 inflammasome including NLRP3,ASC,and pro-caspase-1 in NS rats.SKHGMB reduced the protein and mRNA levels of fibrosis regulators in NS rats.The results indicated that SKHGMB exerts protective effects against renal dysfunction by inhibiting of renal inflammation and fibrosis in NS rats.