Improvement effects of glycyrrhizin on Helicobacter pylori-associated gastritis in rats and its mechanism / 中国药房

OBJECTIVE To investigate the improvement effects of glycyrrhizin （GL） on Helicobacter pylori （HP）-associated gastritis in rats and its mechanism. METHODS HP-associated gastritis rat model was induced by inoculating with 1×109 cfu/mL HP. The model rats were randomly divided into model group， positive control group （HP standard quadruple group）， GL low-dose， medium-dose and high-dose groups （5， 20， 50 mg/kg）， with 12 rats in each group. Another 12 healthy rats were selected as normal control group. Except the normal control group and model group were given constant volume of normal saline intragastrically， the other groups were given corresponding drugs intragastrically， once a day， for 30 consecutive days. After administration， rats received 13C urea breath test， and delta-over-baseline （DOB） was recorded； the pathological and cellular morphological changes of gastric mucosa in rats were observed， and pathological scoring was performed； the levels of interleukin-8 （IL-8）， IL-1β， tumor necrosis factor-α （TNF-α）， reactive oxygen species （ROS） and malondialdehyde （MDA） were detected in gastric mucosa of rats； mRNA expressions of high mobility group box-1 protein （HMGB1） and nuclear factor-κ-B （NF-κB）， relative expressions of nitric oxide synthases （iNOS） and HMGB1， the phosphorylation level of NF- κBp65 were also detected in rats. RESULTS Compared with normal control group， the DOB value， histopathological score of gastric mucosa， the levels of IL-8， IL-1β， TNF-α， ROS and MDA， relative expressions of HMGB1 and NF- κB mRNA， relative expressions of iNOS and HMGB1 protein and the phosphorylation level of NF-κB p65 were all increased significantly in model group （P＜0.05）； the epithelial cells of gastric mucosa in rats were incomplete in structure and decreased in the number， with an increase in cell fragments and vacuoles， and significant cell pyknosis. Compared with model group， the changes of the above indexes in GL groups and positive control group were significantly reversed （P＜0.05）； the changes in the above indicators in the GL high-dose group were more significant than GL low-dose and medium-dose groups （P＜0.05）； the pathological changes of gastric mucosal cells in rats had all improved. CONCLUSIONS GL may inhibit inflammation and oxidative stress by inhibiting the activation of HMGB1/NF-κB pathway， thus relieving HP-induced gastric mucosal injury.

Curcumin plays an anti-osteoporosis role by inhibiting NF-κB signaling pathway to reduce oxidative stress damage to osteogenesis / 中国药理学通报

Aim To investigate the mechanism of curcumin inhibition of oxidative stress on osteogenic differentiation and its dose-dependent anti-osteoporosis effect. Methods Cellular oxidative stress models were used, different concentrations of curcumin were added to determinethebone formation markers, and the potential signaling pathways involvedwere detected. Meanwhile, the mouse model of osteoporosis ( ovariecto- mized, 0VX) was used to confirm its effect against osteoporosis. Results In vitro experiments found that low concentrations of curcumin (1-10 μmol · L

Effect of cangfudaotan decoction on TLR4/NF-κB p65 signaling pathway in obese PCOS-IR rats / 中国药理学通报

Aim To explore the regulatory effect of Cangfudaotan Decoction on the ovarian Toll receptor 4 (TLR4)/nuclear transcription factor kBp65 (NF-κB p65) signaling pathway in obese PCOS-IR rats. Methods Forty-eight female rats were randomly divided into normal group (n = 8) and model group (n = 40). The obese PCOS-IR rats were established by letrozole (1 mg · kg

Mechanism of Flemiphilippinin D Regulating Inflammatory Response in CIA Rats Through TLR2/MyD88/NF-κB Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effect of Flemiphilippinin D on collagen-induced arthritis （CIA） in rats and explore its mechanism. MethodForty rats were randomly divided into normal group， CIA group， methotrexate （MTX） group （1.35 mg·kg-1）， low-dose Flemiphilippinin D group （1.5 mg·kg-1）， and high-dose Flemiphilippinin D group （3.0 mg·kg-1）， with eight rats in each group. Except for the normal group， the CIA model was induced by type Ⅱ collagen. Each group was given corresponding liquid medicine or normal saline， once a week in the MTX group， and once a day in the Flemiphilippinin D groups for a total of 28 days. The arthritis score and joint swelling degree of rats were experimentally recorded. Pathological changes in the ankle joint of rats were observed by hematoxylin-eosin （HE） staining. Serum levels of inflammatory cytokines interleukin （IL）-1β， IL-6， IL-8， and tumor necrosis factor （TNF）-α were detected by enzyme-linked immunoabsorbent assay （ELISA）， and the mRNA expression of Toll-like receptor 2 （TLR2）， myeloid differentiation factor 88 （MyD88）， and nuclear transcription factor-κB （NF-κB） p65 were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and the protein expressions of TLR2， MyD88， and NF-κB p65 were detected by Western blot. ResultCompared with the normal group， the ankle joint of the CIA group was significantly swollen， and the clinical score of arthritis and the degree of joint swelling were significantly increased （P<0.01）. The ankle joint tissue structure was significantly damaged， and the levels of inflammatory factors IL-1β， IL-6， IL-8， and TNF-α in serum were significantly increased （P<0.01）. The mRNA levels and protein levels of TLR2， MyD88， and NF-κB p65 were significantly increased（P<0.01）. Compared with the CIA group， arthritis clinical score and joint swelling of rats in each administration group were significantly reduced （P<0.05， P<0.01）， and the pathological changes in the ankle joint were significantly improved. The contents of serum IL-1β， IL-6， IL-8， and TNF-α were significantly decreased （P<0.05， P<0.01）. The mRNA levels and protein levels of TLR2， MyD88， and NF-κB p65 in the ankle joint were significantly decreased （P<0.05， P<0.01）. ConclusionTo a certain extent， Flemiphilippinin D can reduce the expression of inflammatory factors in rheumatoid arthritis rats and play a good therapeutic effect. It works perhaps by inhibiting the activation of the TLR2/MyD88/NF-κB signaling pathway and thus shows an anti-inflammatory effect.

Effect of Xielitang on TLR4/NF-κB/HIF-1α Signaling Pathway in Mice with Ulcerative Colitis / 中国实验方剂学杂志

ObjectiveTo explore the protective effect of Xielitang on ulcerative colitis （UC） mice induced by dextran sodium sulfate （DSS） and its possible mechanism. MethodSixty C57BL/6 mice were randomly divided into normal group， model group， sulfasalazine group and and low-， medium-， and high-dose Xielitang groups. Free drinking DSS solution to build the chronic UC model mice. Except for normal group， other groups were given 1.5% DSS for 3 cycles of drinking （days 1-7， days 22-28 and days 43-49） and distilled water for the rest of the time （days 8-21， days 29-42 and days 50-63）. After the first cycle， corresponding drugs were given for 42 days. The changes of general condition， body weight and disease activity index （DAI） score of mice were daily recorded during the experiment. At the end of the treatment， serum and colon tissue samples were collected， colon length was measured， intestinal weight index and colonic mucosal injury （CMDI） score were calculated. The pathological status of colon tissue was observed by hematoxylin-eosin （HE） staining. The levels of interleukin-6 （IL-6）， interleukin-10 （IL-10） and tumour necrosis factor-α （TNF-α） were measured by enzyme-linked immunosorbent assay （ELISA）. The gene and protein expressions of Toll like receptor 4 （TLR4）， nuclear transcription factor-κB （NF-κB） and hypoxia inducible factor-1α （HIF-1α） in colon tissue was detected by Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultCompared with the normal group， the body weight， colon length and IL-10 content in the model group were significantly decreased （P<0.01）， DAI score， intestinal weight index， CMDI score， IL-6 and TNF-α contents， and mRNA and protein expression levels of TLR4， NF-κB and HIF-1α in the model group were significantly increased （P<0.01）. Moreover， the structure of colonic mucosa was destroyed and inflammatory cells infiltrated in the model group. Compared with model group， body weight， colon length and IL-10 content in each dose group of Xielitang were significantly increased （P<0.05， P<0.01）， DAI score， intestinal weight index and CMDI score， IL-6 and TNF-α contents， mRNA and protein expression levels of TLR4， NF-κB and HIF-1α were notably decreased （P<0.05， P<0.01）. The pathological injury of colon was obviously alleviated. ConclusionXielitang can significantly improve the inflammatory response of UC mice induced by DSS， and its mechanism may be related to the regulation of TLR4/NF-κB/HIF-1α signaling pathway.

Intervention of Osteoking in Rats with Myofascial Pain Syndrome / 中国实验方剂学杂志

ObjectiveTo clarify the intervention effect of Osteoking （OK） in rats with myofascial pain syndrome （MPS） and preliminarily explore the pharmacological mechanism of OK in relieving chronic pain from the perspective of anti-inflammatory disease. MethodThe 60 SD rats were divided into normal group， model group， low， medium， and high dose OK groups （0.66， 1.31， 2.63 mL·kg-1）， and positive celecoxib group （21 mg·kg-1）. The MPS rat model was established by beating combined with the centrifugal exercise method， and the OK and celecoxib were given at the same time. SMALGO paw pressure pain manometer detected the shock pain point tenderness threshold of rats， and the Von-Frey needle and acetone stimulation method detected the mechanical hyperalgesia threshold and cold hyperalgesia stimulation response respectively. Eight weeks and 10 weeks after modeling， the spontaneous discharge state and convulsion response of MPS rats were determined by electromyograph （EMG） instrument. The gait changes of MPS rats were detected using a CatWalk gait analyzer. The expression levels of interleukin-1 β （IL-1β）， tumor necrosis factor-α （TNF-α）， substance P （SP）， and bradykinin （BK） were measured by enzyme-linked immunosorbent assay （ELISA）. The protein expression levels of nuclear transcription factor-κB （NF-κB） inhibiting protein α （IκBα）， phosphorylates （p）- IκBα， NF-κB p65， and p-NF-κB p65 were detected in MPS rats by Western blot. The positive expression of p-NF-κB p65 was detected by immunofluorescence. ResultCompared with the normal group， the model group shows 100% positive rates for EMG signal and local convulsions response at both the 8th and 10th weeks. The tenderness threshold and mechanical hyperalgesia threshold are significantly reduced. Cold hyperalgesia score is significantly increased， and gait is abnormal. The expression levels of serum and trigger points IL-1β， TNF-α， SP， BK， p-IκBα， and p-NF-κB p65， as well as the positive expression intensity of p-NF-κB p65 are significantly increased （P<0.01）. Compared with the model group， the positive rate of EMG detection and local convulsion response is significantly reduced in the medium and high dose OK groups （P<0.05）. The tenderness threshold and mechanical hyperalgesia threshold increase significantly in the medium and high dose OK groups， and the cold hyperalgesia score is significantly reduced in the high dose OK group （P<0.01）. The standing time， swing time， and walking period are significantly increased. The swing speed， maximum contact area， and maximum contact intensity are significantly decreased in the high dose OK group （P<0.05）. Moreover， the protein expression levels of p-IκBα/IκBα and p-NF-κB p65/NF-κB p65 are significantly reduced in the medium and high dose OK groups （P<0.05，P<0.01）. The positive expression intensity of p-NF-κB p65 is significantly decreased in the high dose OK group （P<0.01）. ConclusionThe mechanism of OK in relieving the pain in trigger points of MPS and improving gait abnormalities is related to the downregulation of the NF-κB p65 inflammatory signaling pathway to reduce the expression of inflammatory factors and pain mediators in blood and trigger point tissue.

Effect of Tanxiang Qingyan Tablets on MyD88/NF-κB/ICAM-1 signaling pathway in rats with chronic bronchitis / 中国药理学通报

Aim To explore the effect of Tanxiang Qingyan Tablets on rat model with chronic bronchitisand the effect of MyD88/NF-κB/ICAM-1 signaling pathway expression in bronchopulmonary tissues of rats.Methods A rat model of chronic bronchitis was established by smoking method combined with lipopolysaccharide(LPS,2g·L-1)tracheal injection.The rats were randomly divided into normal group,sham operation group,model group,and positive drug(Guilong Ruikening tablets)1 g·kg-1)group,Tanxiang Qingyan Tablet high,medium and low dose(1.44,0.72,0.36 g·kg-1)group,with intragastric interventionin continuous 15 days.The pathological changes of bronchopulmonary tissues were observed by HE staining,and the infiltration of bronchial inflammatory cells was counted; ELISA method was used to detect the contents of tumor necrosis factor(TNF-α)and interleukin 10(IL-10)in peripheral serum; Western blot and immunohistochemical methodswere employed to detectmyeloid cell differentiation protein 88(MyD88),nuclear transcription factor-κB(NF-κB)and anti-intercellular adhesion molecule 1(ICAM-1)protein expression in bronchopulmonary tissues.Results Compared with normal group and sham operation group,the bronchial mucosal epithelial cells of model group were severely damaged,the alveolar septum was widened,the bronchial inflammatory infiltrationsignificantly increased,the serum TNF-α levels significantly increased,IL-10 levels decreased, and MyD88,NF-κB and ICAM-1 protein expression levels increased significantly(P<0.05,0.01)in bronchopulmonary tissues; compared with model group,the pathological damage and inflammatory changes of the bronchopulmonary tissues of rats in Tanxiang Qingyan Tablet group were reduced,and the serum TNF-α content was significantly reduced,IL-10 content did not change significantly,and MyD88,NF-κB and ICAM-1 protein expression levels in bronchopulmonary tissues were significantly down-regulated(P<0.05,0.01).Conclusions Tanxiang Qingyan Tablets can effectively improve bronchopulmonary tissue inflammatory infiltration,which may be related to reducing the release of inflammatory mediators such as TNF-α and regulating the expression of MyD88/NF-κB/ICAM-1 signaling pathway.

Effect of Mahonia bealei Leaf Extract on Inflammation in Depression of Rats via NF-κB/NLRP3 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Mahonia bealei leaf extract on depression of rats and the underlying mechanism. MethodThe chemical constituents of the extract were analyzed by HPLC-MS/MS. Forced swimming test and tail suspension test were carried out to estimate the antidepressant effect. The mice were randomly assigned into the following groups： blank group， positive control group （fluoxetine， 10 mg·kg-1）， and Mahonia bealei leaf extract groups （10， 2.5 g·kg-1）. The gavage lasted for 12 days and the immobility time of the mice in the tests was recorded 1 h after the last administration. Furthermore， to explore the underlying mechanism of the antidepressant effect， we established the rat depression model by intraperitoneal injection with reserpine （0.5 mg·kg-1）. Rats were grouped as follows： blank group， model group， positive control group （fluoxetine， 1.8 mg·kg-1）， and Mahonia bealei leaf extract groups （10， 2.5 g·kg-1）. The gavage， once a day， lasted for 10 consecutive days. The depression of rats was detected by behavioral tests 1 h after the last administration. The levels of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in serum of rats were detected by enzyme-linked immunosorbent assay （ELISA）. The protein expression of IL-6 and interleukin-1β （IL-1β） in brain tissue was detected by immunohistochemical staining. The protein levels of nuclear transcription factor-κB （NF-κB） and NOD-like receptor protein 3 （NLRP3） in hippocampus of rats were detected by Western blot. ResultSeven chemical constituents， mainly alkaloids， were identified from the extract. Compared with the blank group， Mahonia bealei leaf extract shortened the immobility time of mice in tail suspension and forced swimming tests. Compared with the blank group， the modeling of rat depression increased the blepharoptosis incidence and retention time in circles （P<0.05， P<0.01）， elevated the levels of IL-6 and TNF-α in serum （P<0.05）， and up-regulated the protein levels of IL-6， IL-1β， NF-κB， and NLRP3 in brain tissues （P<0.01）. Compared with the model group， high dose of Mahonia bealei leaf extract shortened the retention time in circles （P<0.05）， lowered the levels of IL-6 and TNF-α in serum （P<0.05， P<0.01）， and down-regulated the protein levels of IL-6， IL-1β， NF-κB， and NLRP3 （P<0.01） in brain tissues. ConclusionMahonia bealei leaf extract had significant antidepressant effect and alleviated the inflammatory response in reserpine-induced rat model of depression， the mechanism of which may be related to the inhibition of NF-κB/NLRP3 signaling pathway.

Total Glucosides of Paeony Improve Renal Injury in Mice with Systemic Lupus Erythematosus by Regulating TLR9/MyD88/NF-κB Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the intervention effect of total glucosides of paeony （TGP） on the renal injury of MRL/lpr mice based on the Toll-like receptor 9 （TLR9）/myeloid differentiation factor 88 （MyD88）/nuclear transcription factor-κB （NF-κB） signaling pathway and explore the immunological mechanism of TGP in preventing and treating systemic lupus erythematosus （SLE）. MethodMRL/lpr female mice of SPF grade were randomly divided into a model group， a dexamethasone group （0.15 g·kg-1）， and high- （0.078 g·kg-1） and low-dose （0.039 g·kg-1） TGP groups， and female C57BL/6J mice were assigned to a blank group， with 7 mice in each group. Mice in each group were treated with corresponding drugs or normal saline by gavage at the same time every day. After 4 weeks， samples were collected. The kidney and spleen were weighed， and the organ index was calculated. Serum creatinine （SCr） and blood urea nitrogen （BUN） levels in each group were detected by biochemical assay. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in the kidney. The degree of renal fibrosis was evaluated by Masson staining. The serum levels of interleukin （IL）-2， interferon （IFN）-α， IL-4， and anti-nuclear antibody （ANA） were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA expression of TLR9， MyD88， and NF-κB p65 in renal tissues was detected by real-time quantitative polymerase chain reaction （Real-time PCR）. The protein expression of TLR9 and NF-κB p65 in renal tissues was detected by immunofluorescence. The protein expression of TLR9， MyD88， and NF-κB p65 in renal and spleen tissues was tested by Western blot. ResultCompared with the blank group， the model group showed increased SCr， BUN， spleen index， and kidney index （P<0.05）， deteriorated pathological injury and fibrosis in renal tissues， elevated serum levels of IFN-α， IL-4， and ANA， decreased level of IL-2 （P<0.05）， and up-regulated TLR9， MyD88， and NF-κB p65 mRNA and protein levels in the kidney and spleen （P<0.05）. Compared with the model group， the TGP groups displayed reduced SCr， BUN， spleen index， and kidney index （P<0.05）， relieved pathological damage and fibrosis in renal tissues， decreased serum levels of IFN-α， IL-4， and ANA （P<0.05）， increased level of IL-2， and declining mRNA and protein expression levels of TLR9， MyD88， and NF-κB p65 in the kidney and spleen （P<0.05）. ConclusionTGP may inhibit the expression of downstream inflammatory factors to regulate immunity and resist SLE-induced renal injury by regulating the TLR9/MyD88/NF-κB signaling pathway.

Mechanism of Scutellariae Radix-Paeoniae Radix Rubra Combination of Different Proportions Against Hepatic Fibrosis in Rats / 中国实验方剂学杂志

ObjectiveTo observe the effects of Scutellariae Radix （SR）-Paeoniae Radix Rubra （PRR） combination of different proportions on the expression of the Toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear transcription factor κB （NF-κB） and phosphatidylinositol kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathways in liver tissues of rats with hepatic fibrosis and explore the mechanism against hepatic fibrosis. MethodSixty male SD rats of SPF grade were randomly divided into a normal group， a model group， a positive control （silymarin） group， and SR-PRR 1∶1， SR-PRR 1∶2， and SR-PRR 1∶4 groups， with 10 rats in each group. The hepatic fibrosis model was induced in rats except for those in the normal group by intraperitoneal injection of 40% tetrachloromethane （CCl4）-olive oil solution at 3 mL·kg-1， 5 mL·kg-1 for the first time， for 8 weeks， twice per week. After 4 weeks， rats were treated correspondingly at 10 mL·kg-1 by intragastric administration， and the body weight of rats in each group was weighed for 8 weeks. After administration， histopathological changes in the liver were observed. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， hyaluronic acid （HA）， laminin （LN）， albumin （ALB）， alkaline phosphatase （AKP）， and superoxide dismutase （SOD） activities， malondialdehyde （MDA）， and hydroxyproline （HYP） content in liver tissues were detected. The mRNA expression levels of TLR4， MyD88， NF-κB， PI3K， Akt， and mTOR in the liver of rats were detected by real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the model group， SR-PRR combination of different proportions could recover the body weight and improve the pathological injury of the liver. As revealed by enzyme linked immunosorbent assay （ELISA） results， compared with the normal group， the model group showed increased ALT， AST， HA， LN， AKP， MDA， and HYP levels to different degrees （P<0.05）. Compared with the model group， the groups with drug intervention showed decreased levels of ALT， AST， HA， LN， AKP， MDA， and HYP， potentiated SOD activity， and increased level of ALB （P<0.05）. As revealed by Real-time PCR results， compared with the normal group， the model group showed increased mRNA expression of TLR4， MyD88， NF-κB， PI3K， Akt， and mTOR （P<0.05）. Compared with the model group， the groups with drug intervention showed reduced mRNA expression of TLR4， MyD88， NF-κB， PI3K， Akt， and mTOR in the liver of rats （P<0.05）. ConclusionSR-PRR combination of different proportions can improve the histopathological injury in liver tissues caused by CCl4， with the optimal effect observed in the SR-PRR 1∶4 group. SR-PRR may inhibit the development of liver fibrosis by inhibiting the expression of TLR4/MyD88/NF-κB and PI3K/Akt/mTOR signaling pathways， thereby alleviating chemical-induced liver injury.

Qiling Tongluo Prescription Improves Renal Injury in Rats with Idiopathic Membranous Nephropathy by Inhibiting TLR4/MyD88/NF-κB Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore the therapeutic effect and mechanism of Qiling Tongluo prescription against idiopathic membranous nephropathy （IMN） in rats based on Toll-like receptor 4/myeloid differentiation factor 88/nuclear transcription factor-κB （TLR4/MyD88/NF-κB） signaling pathway. MethodSixty male SD rats were randomly divided into the normal group， model group， benazepril hydrochloride （10 mg·kg-1） group， and low-，medium-， and high-dose （6.48， 12.95， and 25.9 g·kg-1） Qiling Tongluo prescription groups. The IMN rat model was established by injection of cationized bovine serum albumin （C-BSA） into the tail vein. After the model was successfully prepared， the rats were gavaged with the corresponding drugs， once a day， for four consecutive weeks. After the treatment， the pathological changes in rat kidneys were observed by hematoxylin-eosin （HE） staining， Masson staining， and periodic acid-silver metheramine （PASM） staining， followed by the detection of 24 h urinary total protein （24 h UTP）， plasma albumin （ALB）， total serum protein （TP）， serum creatinine （SCr）， urea nitrogen （BUN）， and uric acid （UA） levels. The levels of interleukin-1β （IL-1β） and interleukin-6 （IL-6） were determined by enzyme-linked immunosorbent assay （ELISA）， and the mRNA and protein expression levels of TLR4， MyD88， and NF-κB in the kidney tissue were assayed by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， immunohistochemistry （IHC）， and Western blot. ResultCompared with the normal group， the model group exhibited elevated 24 h UTP and serum SCr， BUN， UA， IL-1β， and IL-6 （P<0.05， P<0.01）， decreased ALB and TP （P<0.01）， up-regulated TLR4， MyD88， and NF-κB p65 mRNA and protein expression in kidney tissue （P<0.05， P<0.01）， obvious inflammation， disordered glomerular structure with enlarged volume， irregularly thickened basement membrane， inflammatory cell infiltration in the renal interstitium， reduced renal tubular epithelial cells due to shedding and apoptosis， and some vacuolar degeneration. Compared with the model group， benazepril hydrochloride and Qiling Tongluo prescription at the high dose remarkably lowered the serum SCr and UA （P<0.05） and increased ALB and TP （P<0.05）. Benazepril hydrochloride and Qiling Tongluo prescription at the low， medium， and high doses down-regulated the 24 h UTP， serum IL-1β and IL-6 levels， and renal TLR4， MyD88， and NF-κB p65 mRNA and protein expression to varying degrees （P<0.05， P<0.01）， alleviated IMN inflammatory reaction， glomerular swelling， and volume increase， slightly dilated glomerular capillaries， proliferated mesangial matrix， and relieved pathological and morphological damages in rat kidney， with inflammatory cell infiltration occasionally observed. ConclusionQiling Tongluo prescription may reduce the release and expression of inflammatory factors by regulating the TLR4/MyD88/NF-κB signaling pathway to inhibit the inflammatory response in IMN rats， ameliorate proteinuria and kidney damage， and protect kidney function.

Effect and Mechanism of Xianlian Jiedu Prescription Against Proliferation of Colorectal Cancer Cells in Hypoxic Microenvironment / 中国实验方剂学杂志

ObjectiveTo study the effect of Xianlian Jiedu prescription （XLJDP） on the activation of nuclear transcription factor-κB （NF-κB） signaling pathway induced by bromodomain-containing protein 4 （Brd4） in hypoxic microenvironment and to explore its mechanism in inhibiting the proliferation of colorectal cancer HT-29 cells. MethodThe human colorectal cancer HT-29 cells were cultured in a hypoxic incubator or normoxia incubator and treated with XLJDP at 0.8，1，1.2，1.6，3.2，6.4，and 12.8 g·L-1 for 48 h， respectively. Following the detection of cell vitality using methyl thiazolyl tetrazolium （MTT） colorimetry， the effects of XLJDP （1.25，2.5，and 5 g·L-1） on the cell mitochondrial membrane potential were determined using a fluorescent probe （JC-1）， and the apoptosis of colorectal cancer HT-29 cells was detected by flow cytometry. The cell colony formation assay and 5-ethynyl-2'-deoxyuridine （EDU） staining were conducted to test the proliferation of colorectal cancer HT-29 cells. The Western blot was carried out to measure the expression levels of Brd4 and its downstream relevant proteins such as c-Myc and hexamethylene bisacetamide-inducible protein 1 （HEXIM1）， as well as the effects of XLJDP on related proteins in the NF-κB signaling pathway. ResultCompared with the blank control group， XLJDP at 0.8，1，1.2，1.6，3.2，6.4，and 12.8 g·L-1 inhibited the vitality of colorectal cancer HT-29 cells （P<0.05 ， P<0.01）， with the median inhibitory concentration （IC50） under the hypoxic condition higher than that under the normoxia condition. Compared with the blank control group， XLJDP at 1.25，2.5，and 5 g·L-1 significantly decreased the mitochondria membrane potential， enhanced the apoptosis （P<0.05，P<0.01）， and lowered the number of cell colonies and also the EDU-positive cells （P<0.05， P<0.01）. The results of Western blot showed that compared with the blank control group， XLJDP at 1.25，2.5，and 5 g·L-1 down-regulated Brd4， c-Myc， p-NF-κB p65， and p-IκBα protein expression to varying degrees and up-regulated the expression of HEXIM1 （P<0.05， P<0.01）. ConclusionIn the hypoxic microenvironment， XLJDP inhibits the proliferation of colorectal cancer HT-29 cells regulated by Brd4， which may be related to its inhibition of the activation of NF-κB signaling pathway.

Effect of Wuwei Xiaoduyin on NF-κB Signaling Pathway in Lipopolysaccharide-induced Rat Mesangial Cells / 中国实验方剂学杂志

ObjectiveTo study the effect and mechanism of Wuwei Xiaoduyin in treating rat renal mesangial cells （HBZY-1） induced by lipopolysaccharide （LPS） through the nuclear factor-κB （NF-κB） signaling pathway. MethodRat HBZY-1 cells were randomly assigned into the normal group， model group， benazepril （50 μmol·L-1） group， and high- and low-dose （2.75 and 0.69 g·kg-1） Wuwei Xiaoduyin groups. The normal group， model group， and benazepril group were treated with 10% normal rat serum， and the Wuwei Xiaoduyin groups with 10% medicated serum. Except the normal group， the other four groups were treated with LPS （100 ng·mL-1） for modeling in vitro. The changes of cell morphology were observed under optical microscope. The expression of NF-κB p65 was detected by immunofluorescence （IF） method. Methyl thiazolyl tetrazolium （MTT） colorimetry was employed to detect cytotoxicity and cell proliferation. The levels of interleukin-1β （IL-1β）， intercellular adhesion molecule-1 （ICAM-1）， laminin （LN）， and fibronectin （FN） in cell supernatant were determined by enzyme-linked immunosorbent assay （ELISA）. The mRNA levels of IL-1β， FN， and NF-κB p65 were measured by real-time fluorescence quantitative PCR. The protein levels of phosphorylated inhibitor of NF-κB kinase β （p-IKKβ）， phosphorylated NF-κB inhibitor （p-IκBα）， and NF-κB p65 were determined by Western blot. ResultCompared with the normal group， the modeling increased cell proliferation （P<0.01）， elevated the levels of IL-1β， ICAM-1， LN， and FN in cell supernatant （P<0.01）， and up-regulated the mRNA levels of IL-1β， FN， and NF-κB p65 （P<0.01） and the protein levels of p-IKKβ， p-IκBα， and NF-κB p65 （P<0.01）. Such changes were recovered by benazepril and Wuwei Xiaoduyin （P<0.05， P<0.01）. ConclusionWuwei Xiaoduyin can mitigate the inflammatory injury of renal mesangial cells induced by LPS by inhibiting the NF-κB signaling pathway.

Every-other-day fasting attenuated inflammation in rats after spinal cord injury via the aryl hydrocarbon receptor/suppressor of cytokine signaling 2/nuclear transcription factor-κB signaling pathway / 中国康复理论与实践

ObjectiveTo explore the role of aryl hydrocarbon receptor/suppressor of cytokine signaling 2/nuclear transcription factor-κB (AHR/SOCS2/NF-κB) signaling in the reduction of spinal cord injury (SCI) inflammation with every-other-day fasting (EODF). MethodsA total of 36 healthy male Sprague-Dawley rats were randomly divided into sham group (n = 12), model group (n = 12) and EODF group (n = 12). The latter two groups were established C5 spinal cord hemi-clamp model. The EODF group received EODF, namely fasting and feeding per 24 hours (water free), for two weeks after operation. They were assessed with Grooming Test before operation, and one, seven and 14 days after operationt, respectively. Pathological injury of spinal cord was observed with HE staining 14 days after operation, while the expression of AHR, SOCS2 and NF-κB proteins were detected with Western blotting, and the mRNA of AHR, SOCS2, inhibitor α of NF-κB (IκBα), NF-κB and tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) were measured with reverse transcription real-time quantitative polymerase chain reaction. ResultsCompared with the sham group, the Grooming Test score decreased in the other two groups (P < 0.05), and cavity, edema and structural disorder appeared in the spinal cord. Compared with the model group, the pathology of EODF group improved, and the Grooming Test score increased (P < 0.05). The mRNA and protein expression of AHR and SOCS2 increased (P < 0.05), the mRNA and protein expression of NF-κB decreased (P < 0.05), the mRNA expression of IκBα increased (P < 0.05), and the mRNA expression of TNF-α and IL-6 decreased (P < 0.05). ConclusionEODF may promote the recovery of motor function of forelimb in rats with SCI, and relieve damage and inflammation, which may associate with the activation of AHR/SOCS2/NF-κB signaling pathway.

Effect and Mechanism of Yuxuebi Tablet Against Collagen-Induced Arthritis of Rats / 中国实验方剂学杂志

Objective:To explore the intervention effect of Yuxuebi tablet （YXB） on collagen-induced arthritis （CIA） in rats and its anti-inflammatory mechanism. Method:Following CIA modeling， the rats in the drug administration groups were separately treated with intragastric administration of YXB （0.1， 0.2， and 0.4 g·kg^-1） and methotrexate （MTX， 0.4 mg·kg^-1）， once a day. The incidence of CIA， mechanical pain threshold （MPT） and cold pain threshold （CPT） were evaluated once every three days. After continuous administration for 30 days， the peripheral blood of rats was collected for the determination of platelet （PLT） count and fibrinogen （FIB） content. The hematoxylin-eosin （HE） staining was conducted to analyze the pathological changes in joint tissues. The protein expression levels of interleukin （IL）-1<italic>β</italic>， IL-8， nuclear transcription factor-<italic>κ</italic>B （NF-<italic>κ</italic>B） p65， phosphorylated NF-<italic>κ</italic>B （p-NF-<italic>κ</italic>B） p65， Ras， and Raf-1 in joint tissues of CIA rats were detected by immunohistochemistry （IHC） and Western blot. The rheumatoid arthritis fibroblast-like synoviocytes （RA-FLS） were induced by tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>， 10 μg·L^-1） <italic>in vitro</italic> and then subjected to transwell migration/invasion assay， followed by the detection of protein expression levels of Ras， Raf-1， and p-NF-<italic>κ</italic>B p65 in RA-FLS by Western blot. Result:Compared with the control group， the model group exhibited an increased incidence of CIA， significantly decreased MPT （<italic>P</italic><0.05，<italic>P</italic><0.01）， elevated CPT （<italic>P</italic><0.01） and PLT and FIB in the peripheral blood， worsened histopathological score of joints， enhanced RA-FLS migration and invasion， and up-regulated inflammatory factors （<italic>P</italic><0.01）. The comparison with the model group revealed that YXB at different doses obviously reduced the incidence of CIA， increased MPT， down-regulated CPT and PLT and FIB in the peripheral blood （<italic>P</italic><0.05，<italic>P</italic><0.01）， ameliorated the pathological changes like synovial hyperplasia and bone and cartilage destruction （<italic>P</italic><0.05，<italic>P</italic><0.01）， and inhibited RA-FLS migration and invasion. Besides， the low-， medium-， and high-dose YXB reversed the IL-1<italic>β</italic>， IL-8， Ras， Raf-1， and p-NF-<italic>κ</italic>B p65 expression in joint tissues of CIA rats to different extents， as well as the protein expression of Ras， Raf-1 and p-NF-<italic>κ</italic>B p65 in RA-FLS （<italic>P</italic><0.05，<italic>P</italic><0.01）. Conclusion:YXB reduces the incidence of CIA， ameliorates the clinical symptoms of RA and the pathological changes in joint tissues， and inhibits the formation of synovium， which may be attributed to its inhibition against Ras/Raf-1/NF-<italic>κ</italic>B signaling pathway.

Mechanism of Dahuang Zhechongwan in Inhibiting Silicosis in Mice via p38 MAPK/NF-κB/TGF-β1 Pathway / 中国实验方剂学杂志

Objective:To explore the effects and mechanism of Chinese classical prescription Dahuang Zhechongwan on silicosis in mice. Method:Thirty-six male Kunming mice of SPF grade were randomized into the normal control group， model control group， tetrandrine （Tet， 0.039 mg·kg^-1） group， as well as high- （1.560 g·kg^-1）， medium- （0.780 g·kg^-1）， and low-dose （0.390 g·kg^-1） Dahuang Zhechongwan groups， with six mice in each group. Mice in all groups except for the normal control group underwent static inhalation of silica （SiO₂） dust for 40 consecutive days to induce fibrosis. After 28 days of intervention with corresponding drugs， the mice were sacrificed to collect the serum and lung tissues， with the former used for detecting tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， interleukin-1<italic>β </italic>（IL-1<italic>β</italic>）， IL-6， and hydroxyproline （HYP） levels by enzyme-linked immunosorbent assay （ELISA） and the latter for observing the pathological changes. Meanwhile， the protein and mRNA expression levels of p38 mitogen-activated protein kinase （p38 MAPK）， nuclear transcription factor-<italic>κ</italic>B （NF-<italic>κ</italic>B）， transforming growth factor-<italic>β</italic>₁ （TGF-<italic>β</italic>₁）， <italic>α</italic>-smooth muscle actin （<italic>α</italic>-SMA）， Smad2， Smad3， and Smad7 in the lung tissues were determined by Western blot and real-time polymerase chain reaction （Real-time PCR）. Result:Compared with the normal group， the contents of TNF-<italic>α</italic>， IL-1<italic>β</italic>， IL-6 and HYP in the model group were significantly increased， the difference was statistically significant（<italic>P</italic><0.05，<italic>P</italic><0.01）； compared with the model group， the high-dose group of Dahuang Zhechongwan could significantly reduce the contents of TNF-<italic>α</italic>， IL-6 and HYP in the serum of mice（<italic>P</italic><0.05， <italic>P</italic><0.01）， indicating that Dahuang Zhechongwan could reduce the lung inflammation of silicosis mice. At the same time， compared with the normal group， the protein and mRNA expression levels of p38 MAPK， NF-<italic>κ</italic>B p65， TGF-<italic>β</italic>₁， <italic>α</italic>-SMA， Smad2 and Smad3 in the model group were significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）， while the protein and mRNA expression levels of Smad7 were significantly decreased（<italic>P</italic><0.01）； compared with the model group， the protein and mRNA expression levels of p38 MAPK， NF-<italic>κ</italic>B p65， TGF-<italic>β</italic>₁， <italic>α</italic>-SMA， Smad2 and Smad3 in the high-dose Dahuang Zhechongwan group were significantly increased the protein and mRNA expression levels were significantly decreased（<italic>P</italic><0.05，<italic>P</italic><0.01）， while Smad7 protein and mRNA expression levels were significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）. Conclusion:Dahuang Zhechongwan ameliorates the alveolar inflammation， extracellular matrix （ECM） deposition， and fibrosis in mice with silicosis possibly by regulating the p38 MAPK/NF-<italic>κ</italic>B/TGF-<italic>β</italic>₁ pathway.

Effect of Siwu Paste on Hematopoietic Function of Bone Marrow in Rats with Aplastic Anemia and Its Mechanism / 中国实验方剂学杂志

Objective:To explore the regulatory effect of Siwu paste on the bone marrow hematopoietic function of aplastic anemia （AA） model rats. Method:SD rats were randomly divided into normal control group， model group， positive drug （Fufang E'jiao Jiang 10.8 g·kg^-1） group， high-dose Siwu paste （22.68 g·kg^-1） group and low-dose Siwu paste （5.67 g·kg^-1） group. Acetophenazine （APH） combined with cyclophosphamide （CTX） injection was used to establish the aplastic anemia rat model. The administration groups were given the corresponding drugs （<italic>ig</italic>） for 15 consecutive days. The levels of white blood cells （WBC）， red blood cells （RBC）， hemoglobin （HGB）， hematocrit （HCT） and platelets （PLT） in peripheral blood cells of rats were detected， thymus and spleen indexes were calculated and compared. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin-3 （IL-3） and interleukin-6 （IL-6） in rat serum. The pathological changes of bone marrow were observed by hematoxylin eosin （HE） staining. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） methods were used to detect Toll receptor 4 （TLR4） and nuclear transcription factor-<italic>κ</italic>B （NF-<italic>κ</italic>B） protein and gene expression in rat femoral bone marrow cells. Result:Compared with the normal control group， the WBC， RBC， HGB， HCT and PLT levels of the model group were significantly reduced， the thymus index was significantly decreased， the spleen index was significantly increased， the serum IL-3 level was significantly decreased， and the IL-6 level was significantly increased. The number of neutrophils and megakaryocytes in the femoral bone marrow was reduced， and the medullary cavity was filled with edema fibrofatty tissue. The expressions of TLR4 and NF-<italic>κ</italic>B protein and mRNA in bone marrow cells were significantly increased （<italic>P</italic><0.01）. Compared with the model group， the levels of WBC， RBC， HCT and PLT in peripheral blood cells of rats in the high-dose Siwu paste group increased， the thymus index increased， the spleen index decreased， the IL-3 level was significantly increased， the IL-6 level was significantly decreased， the pathological morphology of femoral bone marrow was slightly improved， and the expressions of TLR4 and NF-<italic>κ</italic>B protein and mRNA in bone marrow cells decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:Siwu paste may improve the bone marrow hematopoietic function of rats with aplastic anemia by regulating the expression of the bone marrow inflammation signal pathway TLR4/NF-<italic>κ</italic>B.

Therapeutic Effect of Guizhi Shaoyao Zhimutang on Osteoporosis in Rats Based on RANKL/OPG / 中国实验方剂学杂志

Objective:To investigate the therapeutic effect of Guizhi Shaoyao Zhimutang （GSZT） on the osteoporosis （OP） in SD female rats induced by ovariectomy （OVX） combined with glucocorticoid injection and its related mechanisms. Method:The rats were divided into a sham operation group and an experimental group for OVX. One week later， the experimental rats were divided into an OP model group， a calciferol （positive control， 0.1 mg·kg^-1） group， and low（0.8 g·kg^-1）， medium（1.6 g·kg^-1）， and high-dose （3.2 g·kg^-1） GSZT groups. Except for those in the sham operation group， the rats received an intramuscular injection of dexamethasone （1 mg·kg^-1） twice per week for six weeks. After the rats were treated correspondingly for eight weeks， the rats were sacrificed for thighbone sample collection. The computer microtomography （Micro CT） was used to analyze the parameters of bone mineral density （BMD）， bone volume （BV）， tissue volume （TV）， structure model index （SMI）， connectivity density （Conn.D）， trabecular number （Tb.N）， trabecular thickness （Tb.Th）， trabecular pattern factor （Tb.Pf）， and trabecular separation （Tb.Sp）. In addition， hematoxylin-eosin （HE） and Masson staining was used for the histopathological examination of the thighbone. Tartrate-resistant acid phosphatase （TRAP） staining was used to analyze osteoclasts in bone tissues. The enzyme-linked immunosorbent assay （ELISA） was used to determine matrix metalloproteinase-9 （MMP-9）， cathepsin K （CTSK）， and TRAP5b in bone tissues， and Western blot was carried out to determine receptor activator of NF-kappaB ligand （RANKL）， osteoprotegerin （OPG）， alkaline phosphatase （ALP） in bone tissues. Result:Compared with normal group， the number of bone trabeculae in cancellous part of OP model rats decreased significantly， the spacing widened， the thickness became thinner， and the continuity was poor. Compared with model group， GZST medium and high-dose groups could improve the changes of femoral bone trabecular cancellous bone. Masson staining showed that compared with the normal group， the growth plate thickness of the distal femur epiphysis was thinner and the area of new bone was smaller in the OP model group， while the reduction of new bone was inhibited in the high dose group of calcitol and GZST. TRAP staining results showed that the number of osteoclasts in OP group was significantly increased compared with the normal group， and the number of osteoclasts in medium-high dose GZST group was reduced. Micro CT results showed that compared with normal group， BMD， BV， Tb.N， Tb.Th and Conn.D of OP rats were decreased， SMI and Tb.Sp were significantly increased（<italic>P</italic><0.01）. Compared with model group， medium and high dose of GZST could improve the degree of osteoporosis in OP rats， BV， Tb.N were significantly increased， SMI， Tb.Sp were significantly decreased （<italic>P</italic><0.01）. ELISA results revealed that compared with the sham operation group， the OP model group showed decreased ALP content （<italic>P</italic><0.01） and increased MMP-9， TRAP5b， and CTSK （<italic>P</italic><0.01）. Compared with the OP model group， the high-dose GSZT group showed increased ALP content （<italic>P</italic><0.01） and decreased MMP-9， TRAP5b， and CTSK （<italic>P</italic><0.01）. Western blot results demonstrated that compared with the OP model group， the calciferol group and the high-dose GSZT group showed inhibited expression of the RANKL protein （<italic>P</italic><0.05）， and increased OPG expression （<italic>P</italic><0.05）. Conclusion:The findings suggest that GZST has a potential therapeutic effect on OP caused by OVX combined with glucocorticoids in rats， and the underlying mechanism is associated with the regulation of RANKL/OPG in bone tissues. This study is expected to lay a preliminary foundation for its clinical application and modern preparation development.

Effects of NF-κB/NOX1 signaling pathway in TNF-α-induced apoptosis of A549 cells / 中国职业医学

OBJECTIVE: To investigate the effects of nuclear transcription factor-κB(NF-κB)/amide adenine dinucleotide phosphate oxidase 1(NOX1) signaling pathway in tumor necrosis factor-α(TNF-α) induced apoptosis of A549 cells. METHODS: i) A549 cells were stimulated with TNF-α at the concentrations of 0.00, 0.25, 0.50, and 1.00 nmol/L. CCK-8 assay was used to detect the cell viability to screen the optimal stimulating concentration of TNF-α. ii) A549 cells at logarithmic growth stage were randomly divided into four groups, the control group, the TNF-α group, the BAY11-7082(NF-κB inhibitor) group and the TNF-α+BAY11-7082 group. The cells in the control group were not treated. The TNF-α and BAY11-7082 groups were stimulated with 0.50 nmol/L TNF-α and 5 μmol/L BAY11-7082, respectively. The TNF-α+BAY11-7082 group was stimulated by both TNF-α and BAY11-7082. After 24 hours of culture, the cell survival rate was detected by CCK-8 assay. Flow cytometry was used to detect cell apoptotic rate, and Western blotting was used to detect the relative expression of NF-κB(p65) and NOX1 proteins. RESULTS: i) When A549 cells were stimulated with TNF-α at the concentration of 0.50 nmol/L, the cell proliferative activity was reduced and the cell apoptosis was promoted. This concentration was selected as the stimulation dose of TNF-α in subsequent experiments. ii) The survival rate of A549 cells in the TNF-α group decreased(P<0.05), the apoptotic rate and the protein expressions of NF-κB(p65) and NOX1 increased in TNF-α group(all P<0.05) compared with the control group. In BAY11-7082 group, the survival rate and the relative expression of NF-κB(p65) and NOX1 of A549 cells were decreased(all P<0.05), and the apoptotic rate of A549 cells was increased(P<0.05) compared with the control group. A549 cells in TNF-α+BAY11-7082 group changed from a long spindle shape to an irregular one. The cell survival rate increased(P<0.05), the apoptotic rate and the relative expression of NF-κB(p65) and NOX1 decreased(all P<0.05) compared with the TNF-α group. CONCLUSION: NF-κB/NOX1 signaling pathway is involved in A549 cells apoptosis induced by TNF-α.

Effect of Testudinis Carapax et Plastrum Aqueous Extract Regulating NF-κB Inflammation Microenvironment on Osteogenic Differentiation of MC3T3-E1 / 中国实验方剂学杂志

Objective::To investigate the role and mechanism of Testudinis Carapax et Plastrum aqueous extract in promoting osteogenic differentiation of mouse preosteoblast cell line(MC3T3-E1) by regulating nuclear transcription factor-κB(NF-κB) inflammation microenvironment. Method::MC3T3-E1 cells were cultured in vitro, and osteogenic induction (OI) was performed. Testudinis Carapax et Plastrum was prepared and treated the cells. Cells were devided into control group, osteogenic induction group and Testudinis Carapax et Plastrum (20 mg·L-1)with osteogenic induction group. The proliferation of MC3T3-E1 was detected by cell counting kit-8 (CCK-8), and the optimum concentration of intervention was determined. MC3T3-E1 differentiation and osteogenic mineralization were assayed using alkaline phosphatase (ALP) and Alizarin red staining (ARS), respectively. The expressions of NF-κB p65, NF-κB p105, interleukin-6(IL-6), ALP and Collagen-Ⅰ(COL-Ⅰ) mRNA were detected by Real-time PCR. Result::The results of CCK-8 showed that the proliferation of MC3T3-E1 did not change statistically with time, but it showed an upward trend, while the proliferation at 20 mg·L-1 was more obvious than other groups. The ALP and ARS showed that the positive staining rate of osteogenic induction group and Testudinis Carapax et Plastrum with osteogenic induction group were higher than control group.Real-time PCR results showed that on the 7th day in culture, the expression of NF-κB p105 and IL-6 mRNA in Testudinis Carapax et Plastrum with osteogenic induction group was significantly lower than that in control group (P<0.01), and the expression of ALP and COL-Ⅰ mRNA was significantly upregulated(P<0.05), on the 14th day, the expression of NF-κB p65, NF-κB p105 and IL-6 mRNA in Testudinis Carapax et Plastrum with osteogenic induction group was significantly lower than that in control group (P<0.01). The expression of ALP and COL-Ⅰ mRNA was significantly increased (P<0.05, P<0.01). Conclusion::Testudinis Carapax et Plastrum aqueous extract can promote osteogenic differentiation of MC3T3-E1 via a mechanism associated with the regulation of inhibition of NF-κB inflammatory microenvironment.