经皮耳迷走神经刺激干预功能性消化不良模型大鼠的效应及机制研究

ObjectiveTo explore the effect and its possible mechanism of the intervention of transcutaneous auricular vagus nerve stimulation (taVNS) for the functional dyspepsia (FD) model rats.MethodsOf the 25 male SD rats, 6 rats were set as blank group, the other 19 rats were established to be the functional dyspepsia (FD) model by iodoacetamide intragastric administration, and 18 FD models were successfully established. The 18 model rats were randomly divided into a model group, a sham-taVNS group, and taVNS group, with 6 rats in each group. There was no intervention applied to the rats in the blank and model groups. Whereas, the rats in sham-taVNS group received stimulation to the rim of auricular concha of both sides, and those in taVNS group received stimulation to the cavity of auricular concha of both sides, for 30 min each time, once a day, 7 days in succession. After the intervention, the gastric sensitivity of the rats in each group under different pressure conditions in the stomach, the expressions of serum brain-gut peptide motilin (MTL), cholecystokinin (CCK), glucagon-likepeptide1 (GLP-1), and inflammatory factors IL-4, IL-10, and IL-1β were detected.Results(1) Gastric sensitivity: compared with the blank group, the gastric sensitivity of the model group was higher (P <0.05). Compared with the model group, the gastric sensitivity of the taVNS group was lower (P < 0.05). Compared with the sham-taVNS group, the gastric sensitivity of the taVNS group was lower (P <0.05).(2) Expression of brain-gut peptide: compared with the blank group, MTL was lower, CCK and GLP-1 were higher in the model group (all P <0.05). Compared with the model group, MTL was higher, CCK and GLP-1 were lower in the taVNS group (all P <0.05). Compared with the sham-taVNS group, CCK and GLP-1 were lower in the taVNS group (both P<0.05). (3)Expression of inflammatory factors: compared with the blank group, IL-4 and IL-10 were lower and IL-1β was higher in the model group (all P <0.05). Compared with the model group, IL-10 was higher and IL-1β was lower in the sham-taVNS group (all P <0.05), while IL-4 and IL-10 were higher and IL-1β was lower in the taVNS group (all P <0.05). Compared with the sham-taVNS group, IL-4 and IL-10 were higher and IL-1β was lower in the taVNS group (all P <0.05).ConclusionTaVNS can reduce the gastric sensitivity of FD model rats by peripheral anti-inflammatory action and regulating the abnormal secretion of brain-gut peptide.     

Intervention of the Mahuang Lianqiao Chixiaodou decoction on immune imbalance in atopic dermatitis-like model mice

ObjectiveTo explore the potential mechanism of intervention on the immune imbalance of atopic dermatitis (AD) by studying the effects of Mahuang Lianqiao Chixiaodou decoction (MLCD) on skin damage and inflammation factors in an AD-like mouse model.MethodsNinety-six male BALB/c mice were divided into normal, model, positive control (mometasone furoate), and traditional Chinese medicine treatment (MLCD) groups by a random number table. 2,4-dinitrofluorobenzene was used to induce AD-like mice in all groups except the normal group. The treatment or intervention was administered for seven consecutive days on days 4, 18, 32, and 39. The mRNA relative expressions of interleukin-4 (IL-4), IL-10, interferon-γ (IFN-γ), thymic stromal lymphopoietin (TSLP), and the TSLP receptor (TSLPR) were measured using quantitative real-time polymerase chain reaction, and the serum immunoglobulin E, IL-4, IL-10, and IFN-γ levels were detected using enzyme-linked immunosorbent assay.ResultsCompared with the normal group, the hematoxylin-eosin staining of the skin lesions of the mice in the model group was significantly thickened on days 11, 25, and 39. Compared with the model group, the epidermal thickness of the positive control group was significantly alleviated on day 39 (P < .001), and that of the MLCD group was significantly improved on days 25 and 39 (P < .001). Compared with the four observation time points, MLCD had the best treatment effect on day 39 of the experiment and significantly improved the skin damage performance and relieved pathological lesions. On day 39, compared with the model group, MLCD downregulated the skin mRNA relative expressions of IL-4 (P = .009), TSLP (P = .030), and TSLPR (P < .001), and reduced the mouse serum levels of IL-4 (P = .003). For other serum indicators, no significant difference was observed between the model and MLCD groups.ConclusionMLCD improved AD-like mice skin damage by regulating the Th1/Th2 immune imbalance.     

Hewei Jiangni granule alleviates visceral hypersensitivity in a rat model of non-erosive reflux disease via transient receptor potential channel signaling

ObjectiveTo uncover the underlying mechanism of Hewei Jiangni granule (HWJNG) on non-erosive reflux disease (NERD) treatment by examining histological changes, gastrointestinal neurochemicals release and visceral hypersensitivity-related receptor expression in NERD model rats.MethodsA NERD rat model was established via a combination of basal sensitization and acid perfusion. HWJNG treatments at different doses were then administered. Pathological changes to tissues, mast cell (MC) activation, serum levels of esophageal visceral hypersensitivity-related neurochemicals, and transient receptor potential (TRP) receptor mRNA and protein levels were investigated.ResultsCompared with the control group, the expression of tryptase in MCs, the changes of intercellular space, and the serum levels of substance P (SP), calcitonin gene-related peptides (CGRP) and proteinase-activated receptor 2 (PAR2) increased in the model group (all P < .05). The expression of TRP vanilloid 1 (Trpv1) mRNA decreased in esophagus and dorsal root ganglia (DRG) of the model group (P = .030 & P = .013), and the expression of TRP melastatin channel subfamily member 8 (Trpm8) mRNA decreased in the esophagus of model group (P < .01). The level of esophageal TRPV1 protein increased in the model group (P < .01) and the level of TRPM8 protein decreased in esophagus and DRG of the model group (both P < .05). Compared with the model group, the serum levels of SP, CGRP, and PAR2 in the medium-dose HWJNG group showed significant decreases (all P < .05). The expression of Trpv1 mRNA in esophagus and DRG of the HWJNG groups and the Omeprazole group remarkably decreased (all P < .05), as was the expression of Trpm8 mRNA in esophagus of the HWJNG groups (all P < .05).ConclusionHWJNG alleviated visceral hypersensitivity in NERD model rats by regulating TRP-mediated signaling. Our results indicate that HWJNG has potential as a therapeutic agent for NERD.     

Hemostatic mechanism of the effect of Jianpi Yiqi Shexue decoction on vascular factors in immune thrombocytopenia model mice

ObjectiveTo explore the hemostatic mechanism of Jianpi Yiqi Shexue decoction (JYSD) by regulating vascular factors in an immune thrombocytopenia (ITP) mouse model.MethodsAn ITP mouse model was established by the passive-immune modeling method, and interventional drugs used were prednisone tablets and JYSD. The platelet count; vascular activity–related factors vWF, VCAM-1, and TM; and VEGF and bFGF were used as observational indicators.ResultsOn the 8th day of administration, compared with the model group, platelet counts in the prednisone and JYSD groups increased (both P < .001). Compared with the control group, the levels of vWF, VCAM-1, and TM in the other groups were lower (all P < .05). The VCAM-1 level in the JYSD group was higher than that in the prednisone group (P = .012), but without significant difference compared with the model group (P = .051). The TM level in the JYSD group was the lowest (vs. the model group, P = .047; vs. the prednisone group, P = .006). Compared with the control group, the IOD values of VEGF and bFGF in the other three groups were lower (all P < .01). The IOD values of VEGF in the prednisone and JYSD groups were both higher than those in the model group (P = .002 and P < .001, respectively). The IOD values of bFGF among the model, prednisone, and JYSD groups were not statistically significant (P > .05).ConclusionA vascular factor disorder is involved in the pathogenesis of ITP. JYSD can increase the platelet count, upregulate VEGF expression, and reduce the TM level. JYSD has the same effect as prednisone tablets in regulating platelet, vWF, VEGF, and bFGF, with a stronger effect in normalizing VCAM-1 and TM levels. The hemostatic mechanism of JYSD is closely related to the effective balance of vascular factors.     

Xiaoqinglong decoction reduces dendritic cell differentiation and regulates the Th1/Th2 balance in a mouse model of allergic asthma

BackgroundXiaoqinglong decoction (XQLD) is a classic Chinese medicinal formula that is widely used to treat allergic asthma. Recently, the use of XQLD to treat allergic asthma has inspired research to determine its mechanism of action. Because dendritic cells (DCs) and the T helper 1 (Th1)/Th2 cytokine balance play important roles in allergic asthma, the present work aimed to assess how these immune system components are affected by XQLD.MethodsThirty-six female BALB/C mice were randomly divided into three groups: an ovalbumin-based allergic asthma model group, a XQLD treatment group, and a control group. Histology was performed with haematoxylin and eosin staining and immunohistochemical staining. Bronchoalveolar lavage fluid and blood were collected from the animals and used to analyze the composition of inflammatory cells and expression levels of the cytokines interleukin (IL)-5 and IL-13. The thymic stromal lymphopoietin (TSLP) protein expression was assessed by western blot analysis, and the Gata3 and Tbx21 mRNA levels were assessed by polymerase chain reaction.ResultsCompared with the OVA group, the levels of TSLP expression, IL-5, IL-13, and immunoglobulin E in the XQLD group were lower (all P < .01). The level of IL-4-expressing cells (Th2 cells) was lower (P = .0013), and the percentage of IFN-γ-expressing cells (Th1 cells) was higher in the XQLD group compared with those in the OVA group (P = .0065). In addition, XQLD increased the expression of Tbx21 mRNA and decreased the expression of Gata3 mRNA in the lungs compared with the OVA group (both P < .01).ConclusionThese findings suggest that XQLD may ameliorate the course of allergic asthma by regulating the Gata3/Tbx21 balance and inhibiting TSLP expression, changes which are indicative of an altered Th1/Th2 balance. Thus, the clinical effectiveness of XQLD in treating allergic asthma may be due to its regulation of Th1/Th2 balance.     

Integrated network pharmacology and experimental verification to explore the mechanism of Sangqi Qingxuan formula against hypertensive vascular remodeling

ObjectiveTo investigate the bioactive components of Sangqi Qingxuan formula (SQQX), predict the pharmacological targets, and explore the mechanism of hypertensive vascular remodeling (HVR).MethodsNetwork pharmacology was adopted to predict how SQQX acts in HVR. The effectiveness was assessed by blood pressure measurements and pathological morphology observation based on a spontaneously hypertensive rat model, while the mechanism of SQQX on HVR was validated by immunohistochemistry (IHC) and western blot (WB) according to the results of network pharmacology.ResultsThere were 130 bioactive components of SQQX and 231 drug targets predicted by the Traditional Chinese Medicine Systems Pharmacology Database. Subsequently, 181 common targets were identified for SQQX against HVR, with TP53, MAPK1, and AKT1 as the core targets. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses was employed to identify the top 20 enriched functions and the top 20 pathways (P < .01). Finally, the key role of the ERK/MAPK signaling pathway in HVR was determined. The in vivo results suggested that SQQX reduced systolic blood pressure and increased the ratio of thoracic aortic wall thickness to lumen diameter. Additionally, compared with the model group, SQQX increased the expression of smooth muscle 22 alpha (IHC: P < .001; WB: P < .05) and decreased the expression of osteopontin (IHC: P < .001; WB: P < .05), ERK1/2 (IHC: P < .001; WB: ERK1 & ERK2, all P < .05), p-ERK1/2 (IHC: P < .001; WB: ERK1 & ERK2, all P < .05), and the ratio of p-ERK1/2 to ERK1/2 protein (IHC: P < .001).ConclusionsSQQX, which has multiple bioactive ingredients and potential targets, is an effective treatment for HVR. The mechanism of antihypertensive and vascular protection may be related to the inhibition of phenotypic transformation of vascular smooth muscle cells and the ERK/MAPK signaling pathway.     

Seasonal photoperiodic influence of pineal melatonin on hypothalamic-pituitary-adrenal axis-hippocampal-receptor in male rats

BackgroundBased on the effect of seasonal changes on human visceral function, this study investigated the impact of seasonal photoperiod of the pineal body on hypothalamic-pituitary-adrenal axis-hippocampal-receptor in rats, aiming to reveal the mechanism by which pineal gland melatonin regulates the seasonal secretion of hippocampal neurotransmitters.MethodsVernal equinox, summer solstice, autumn equinox, and winter solstice were selected as four experimental time points, and rats were randomly divided into normal control group, sham operation group, and pinealectomized group. The seasonal changes in corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), corticosterone, hypothalamic melatonin receptor (MTR), and hippocampal corticosterone receptor (CORTR) were examined by enzyme-linked immunosorbent assay.ResultsComparing the same group between different seasons, we showed that in the normal control group, CRH, ACTH, corticosterone, and MTR were higher, while CORTR was lower in autumn and winter than in spring (all P < .05). Compared with the normal control group, the pinealectomized group showed higher levels of corticosterone (P = .01), MTR (P = .01), and CORTR (P = .03) during spring; reduced levels of MTR and CORTR (both P < .001) during summer; higher levels of ACTH (P = .001) and MTR (P < .001), and lower levels of CRH (P = .001), corticosterone (P < .001), and CORTR (P = .003) during autumn; and lower levels of CRH (P < .001) and MTR (P = .004), and higher level of ACTH (P < .001) in winter.ConclusionsSeasonal photoperiod acts on the pineal gland to secrete different levels of melatonin, resulting in seasonal changes in the hypothalamic-pituitary-adrenal axis-hippocampal-receptor, which may be the pathophysiological basis for the onset of seasonal affective disorder.     

Effects of electroacupuncture combined with hydrogel on the formation and changes in the glial scar in rats with spinal cord injury

ObjectiveTo observe the effect of electroacupuncture (EA) combined with oriented conductive bioprotein hydrogel (OCBH) on the recovery of nerve function in rats with complete spinal cord injury (SCI) and to explore its effect and mechanism on the formation and changes of glial scars.MethodsA total of 72 female Sprague–Dawley rats were randomly divided into groups according to the treatment received. A rat model of complete SCI was constructed using a spinal cord transection. Behavioral assessments, hematoxylin-eosin (H&E) staining, immunofluorescence staining, and Western blotting were performed at a fixed period after the operation.ResultsThe material group and the material + EA group obtained better results in the behavioral assessments (all P < .05) and the H&E staining. In the immunofluorescence staining and Western blotting, the GFAP protein was expressed more and denser in the material group and the material + EA group than in the model group, and the density of the GFAP expression in the material + EA group was lower at week 12 than in the material group (all P < .05). The expression of complement C3 in the model, material, and material + EA groups decreased in turn. Some inflammatory factors and the NF-κB signaling pathway showed similar results in the Western blotting (all P < .05). The expression of the GDNF protein in the material + EA group was significantly higher than that in the model group and the material group (both P < .01).ConclusionEA combined with OCBH can promote the recovery of motor functions after SCI by facilitating the formation of glial scars in the early stage, preventing the further spread of an inflammatory response that would affect the activation of A1/A2 astrocytes and change the morphology of glial scars at the spinal cord–material interface in its late stage.     

Effects of Xiangsha Liujunzi decoction drug serum on gastric antrum smooth muscle cells from rats with functional dyspepsia by regulating gastrointestinal hormones

ObjectiveTo observe the effect and mechanism of Xiangsha Liujunzi decoction (XSLJZD) drug serum on gastric antrum smooth muscle cells (SMCs) in rats with functional dyspepsia (FD).MethodsGastric antrum SMCs from rats with FD were isolated, cultured, and then divided into six groups as follows: control, model, domperidone, low-dose XSLJZD (LXSLJZD), medium-dose XSLJZD (MXSLJZD), and high-dose XSLJZD (HXSLJZD). Each group was administered the corresponding drug serum for intervention. Drug serum intervention conditions and proliferative activity of SMCs were tested by cholecystokinin octapeptide. Ghrelin, gastrin, somatostatin, and substance P (SP) levels were measured by ELISA. Somatostatin and SP mRNA expression was measured by real-time PCR.ResultsA concentration of 10% drug serum for 24 h was decided to be the best intervention condition for later study. The mean optical density value in the model group was lower than that in the control group (P = .001). Optical density values in the domperidone and HXSLJZD groups were higher than those in the model group (P = .025, P = .032, respectively). Gastrin, SP, and ghrelin levels in the model group were lower (P = .007, P = .037, P = .005, respectively), but somatostatin levels were higher, compared with those in the control group (P = .031). Gastrin, SP, and ghrelin levels in the domperidone, MXSLJZD, and HXSLJZD groups were higher than those in the model group (all P<.05). Somatostatin levels in the four drug-treated groups were lower than those in the model group (P = .002, P = .007, P = .001, P = .009, respectively). SP mRNA levels in the model group were lower than those in the control, domperidone, MXSLJZD, and HXSLJZD groups (P = .037 P = .016, P = .025, P = .002, respectively). Somatostatin mRNA levels in the model group were higher than those in the control and MXSLJZD groups (P = .042, P = .035).ConclusionsXSLJZD and domperidone drug serum effectively promote proliferative activity of gastric antrum SMCs in an FD model. The mechanism of this activity may be regulated by gastrointestinal hormones.     

柴胡加龙骨牡蛎汤对心肌梗死合并焦虑大鼠海马区NLRP3/GSDMD炎性信号通路的影响

目的探讨心肌梗死合并焦虑大鼠海马区NLRP3/GSDMD炎性信号通路变化及柴胡加龙骨牡蛎汤对大鼠海马区炎性损伤的作用效应。方法将60只SD大鼠随机分为假手术组9只、心梗并焦虑组17只、心梗组17只、心梗并焦虑+中药组17只。心梗组采用结扎冠状动脉左前降支方法进行造模;假手术组穿线不结扎;心梗并焦虑组和心梗并焦虑+中药组先采用空瓶刺激方法进行焦虑造模(周期21 d),于造模第15天采用结扎冠状动脉左前降支方法进行心肌梗死造模。心肌梗死造模后第2天,心梗合并焦虑+中药组给予柴胡加龙骨牡蛎汤10 mL/kg灌胃,其余组灌胃等量蒸馏水,均1次/d,连续7 d。灌胃结束后进行心脏超声检查,采用高架十字迷宫实验、旷场实验评价各组大鼠行为学,大脑海马区HE、尼氏染色观察脑神经元病理改变,免疫组化检测大鼠海马区NLRP3、Caspase-1、GSDMD表达情况,Western blot法检测大鼠海马组织中NLRP3、Caspase-1、GSDMD、IL-1β、IL-18蛋白表达水平。结果与心梗并焦虑组比较,心梗并焦虑+中药组左室射血分数(LVEF)、左室短轴缩短率(LVFS)均明显升高(P均<0.05),左心室舒张末期内径(LVIDd)和左心室收缩末期内径(LVIDs)均明显缩短(P均<0.05),大鼠进入开放臂次数所占比和开放臂停留时间所占比、大鼠水平运动得分和垂直运动得分均明显升高(P均<0.05)。心梗并焦虑组和心梗组大鼠海马区尼氏阳性面积均明显低于假手术组(P均<0.05),心梗并焦虑+中药组明显高于心梗并焦虑组和心梗组(P均<0.05)。心梗并焦虑组和心梗组大鼠海马区NLRP3、Caspase-1、GSDMD表达IOD值和海马组织中NLRP3、Caspase-1、GSDMD、IL-1β、IL-18蛋白表达水平均明显高于假手术组(P均<0.05),且心梗并焦虑组GSDMD表达IOD值和NLRP3、Caspase-1、IL-18蛋白表达水平均明显高于心梗组(P均<0.05),而心梗并焦虑+中药组NLRP3、Caspase-1、GSDMD表达IOD值和NLRP3、Caspase-1、GSDMD、IL-1β、IL-18蛋白表达水平均明显低于心梗并焦虑组(P均<0.05)。结论柴胡加龙骨牡蛎汤可能通过调控海马区NLRP3/GSDMD炎性信号通路,抑制心肌梗死合并焦虑大鼠海马区的炎性反应,缓解大鼠焦虑状态。     

当归多糖对糖尿病肾病KK-Ay小鼠肾脏AMPK信号通路及线粒体自噬的影响

目的 研究当归多糖对糖尿病肾病（diabetic nephropathy,DN）KK-Ay小鼠肾脏磷酸腺苷激活的蛋白激酶（AMPactivated protein kinase,AMPK）信号通路及线粒体自噬的影响。方法 SPF级雄性KK-Ay小鼠用高糖高脂饲料喂养,随机分为模型组、厄贝沙坦（25 mg/kg）组和当归多糖高、中、低剂量（400、200、100 mg/kg）组,每组10只;将10只雄性C57BL/6J小鼠作为对照组。给予药物干预4周,观察小鼠一般情况,每周称定体质量并检测血糖;末次给药后,心脏取血并处死小鼠,分离血清检测尿微量白蛋白（urine microalbuminuria,U-ALB）、肌酐（creatinine,SCr）、尿素氮（urea nitrogen,BUN）;采用苏木素-伊红（HE）染色观察肾组织病理变化;采用Western blotting检测肾组织线粒体自噬相关蛋白[微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)、p62、Nix]和线粒体裂变蛋白[线粒体动力相关蛋白1(dy...     

Shenkang Injection protects against diabetic nephropathy in streptozotocin (STZ)-induced mice through enhancement of anti-oxidant and anti-inflammatory activities

ObjectiveTo investigate the protective effects and possible mechanisms of Shenkang Injection (SKI) on the diabetic nephropathy in streptozotocin-induced mice.MethodsSTZ with the feeding of high fat diet (HFD) was used to induce diabetic mice. The balb/c mice and diabetic mice were then randomly divided into five groups: (1) control group, (2) model group, (3) alprostadil (Alp, 1.5 μg/kg) group, (4) SKI (30 ml/kg) group, (5) Alp (1.5 μg/kg) + SKI (15 ml/kg) group. After six weeks' treatment, blood, urine and kidney tissues were collected for biochemical assay, ELISA assay, and pathological analysis.ResultsDiabetic mice exhibited evident manifestations of diabetic nephropathy (DN), as indicated by increased 24-h urine volume, urinary albumin and kidney weight index (P < 0.01), which could be attenuated by SKI treatment (P < 0.01). SKI was further found to improve abnormal morphology in glomerulus with increased glomerular volume and to decrease urinary N-acetyl-b-D-glucpsaminidase (NAG), β2-microglobulin (β2-MG), and kidney injury molecules-1 (KIM-1) levels (P < 0.05, P < 0.01). Plasma levels of anti-oxidant enzymes significantly reduced in the diabetic mice, and those decreases could be reversed by SKI and Alp treatments. Additionally, SKI obviously suppressed the diabetes-induced increases of pro-inflammatory cytokines (IL-6, IL-1β and TNF-α) (P < 0.01). Meanwhile, SKI was found to effectively attenuate the diabetes-induced coagulation dysfunction, as evidenced by lengthening prothrombin and thrombin time, and decreasing plasma levels of fibrinogen (FIB), 6-K-PGF1α and thromboxane B2 (TXB2) (P < 0.05, P < 0.01). With SKI and Alp combined treatment, the anti-oxidant activities and improvements of coagulation dysfunction were enhanced.ConclusionSKI possesses a remarkable property to prevent diabetic nephropathy. The improvements of kidney function and hypercoagulability by SKI were enhanced with Alp combined treatment. The molecular mechanisms underlying the protection of SKI against DN may be related to enhancing the anti-oxidant and anti-inflammatory activities, and improving the coagulation dysfunction.     

Effect of acupotomy on chondrocyte proliferation and expression of CyclinD1, CDK4 and CDK6 in rabbits with knee osteoarthritis

ObjectiveThe aim of this study was to investigate the mechanism of acupotomology (Apo) in the prevention of articular cartilage destruction via the promotion of chondrocyte proliferation and chondrocyte expression of cell cycle regulators, CyclinD1, CDK4 and CDK6 in a rabbit knee osteoarthritis (KOA) model.MethodsTwenty-eight rabbits were randomly divided into a control group, an OA (osteoarthritis) model group, an Apo (acupotomology) group and EA (electro-acupuncture) group (n = 7). Improved Videman's method was used to induce a rabbit model of KOA over 6 weeks. One week later, acupotomy and electro-acupuncture therapy was applied to animals in the respective groups and treatment lasted 4 weeks. Following these treatments, quantitative real-time PCR, immunohistochemical staining and western blotting were performed to assess the mRNA and protein levels of cell cycle regulators CyclinD1 (Cell cycle protein D1), CDK4 (Cyclin-dependent kinase 4) and CDK6 (Cyclin-dependent kinase 6). Ethology measures and knee morphology were also compared among groups.ResultsThe Lequesne MG index score of morphology was increased (P < .01), and the passive range of motion (PROM) and the mRNA and protein levels of CyclinD1, CDK4, and CDK6 were significantly decreased (P < .01) in the OA model compared with the control group. The Lequesne MG index score and the morphology score were decreased in the Apo and EA group compared with the OA model group (P < .05 or P < .01), while the mRNA levels of CyclinD1, CDK4, and CDK6, and the protein levels of CDK4 were increased in the Apo and EA groups compared with the OA model group (P < .05 or P < .01). The PROM, and the protein levels of CyclinD1 and CDK6 were increased (P < .05) in the Apo group compared with the OA model group, while the PROM and the protein levels of CyclinD1 and CDK6 in the EA group were not significantly different (P > .05). Compared with the EA group, the morphology score was decreased in Apo group (P < .05).ConclusionsThe mRNA levels of CyclinD1 and CDK4, and the protein level of CDK4 in chondrocytes are regulate by both Apo and EA. Apo is more effective than EA in regulating the protein levels of CyclinD1 and CDK6. According to the observed changes in morphology and cytokine levels, acupotomy can promote chondrocyte proliferation and can alleviate the destruction of articular cartilage in a model of KOA.     

Assessment of traditional Chinese medicine pattern in a bleomycin-induced pulmonary fibrosis mouse model: A pilot study

ObjectiveTo initially explore traditional Chinese medicine patterns in a bleomycin-induced pulmonary fibrosis mouse model.MethodsThirty-six C57BL/6 mice were divided by the random number table method (with 12 rats per group) into three groups: a blank group, a model group, and a number 2 Feibi recipe (FBR-2) group. The pulmonary fibrosis mouse model was established by intratracheal instillation of bleomycin. The FBR-2 group was treated with FBR-2 for 4 weeks. Symptoms in the mice such as mental behavior, food/water intake, body weight, body temperature, respiratory rate, and tongue image were observed. The samples were collected on the 14th day and 28th day after modeling, and lung tissues were visually assessed and microscopically evaluated by staining with hematoxylin-eosin and Masson. The expression levels of hydroxyproline, interleukin (IL)-33, IL-37, tissue plasminogen activator, and plasminogen activator inhibitor-1 were determined by enzyme-linked immunosorbent assay.ResultsMice in the model group were poor in spirit, less active, slow in response, showed reduced food/water intake, body temperature, and body weight, increased respiratory rate, and their tongue color had changed from light red to dark red. However, treatment with FBR-2 significantly improved these symptoms. Extensive inflammatory cell infiltration and collagen fiber deposition were observed in the lung tissues of the model group. Compared with the blank group, the levels of hydroxyproline, IL-33, and plasminogen activator inhibitor-1 in the model group significantly increased (all P < .05), whereas that of tissue plasminogen activator significantly decreased on the 14th day and 28th day (P = .036 and P = .005, respectively). Moreover, FBR-2 improved lung inflammation and fibrinolysis imbalance and reduced collagen fiber deposition.ConclusionTo some extent, our bleomycin-induced pulmonary fibrosis mouse model exhibited traditional Chinese medicine patterns of qi deficiency, blood stasis, and heat retention.     

Ginsenoside Rb1 attenuates lipopolysaccharide-induced chronic neuroinflammation in mice by tuning glial cell polarization

ObjectiveTo evaluate whether ginsenoside Rb1 (Rb1) can attenuate lipopolysaccharide (LPS)-induced chronic neuroinflammation in mice and to explore its relationship with glial cell polarization.MethodsIntraperitoneal injection with an escalating dose of LPS was used to establish a chronic neuroinflammation model in mice. Once LPS was initiated, 10 or 20 mg/kg Rb1, or sterile saline, was administered for 14 consecutive days. Open field test and beam walking test were used to monitor the changes in behavior. The concentration of cytokines in the serum and brain were used to monitor the systemic inflammation and neuroinflammation, respectively. Molecules specific to each glial cell phenotype were used to investigate glial cell polarization.ResultsMice in the LPS group had reduced spontaneous activities and impaired beam walking performance. Rb1 obviously eased LPS-induced behavior disturbances. Regarding the levels of serum cytokines, both tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were significantly increased, while interleukin-10 (IL-10) and transforming growth factor β (TGF-β) remarkably decreased after LPS treatment (all P < .001). Rb1 treatment significantly attenuated LPS-induced serum cytokine changes (all P < .05). The results of quantitative polymerase chain reaction and western blotting showed that the mRNA and protein expression levels of TNF-α and complement component 3 (C3) in the brain were significantly increased after LPS treatment (all P < .01). Rb1 treatment significantly inhibited LPS-induced inflammation in the brain (all P < .05). Glial cell polarization analysis showed that M1 and M2 microglia, and A1 astrocytes increased following LPS treatment, while A2 astrocytes decreased. Rb1 treatment reduced M1 and M2 microglia, and A1 astrocytes, and significantly increased A2 astrocytes.ConclusionRb1 can attenuate chronic neuroinflammation induced by LPS in mice, which may be partially attributable to its fine tuning of microglia and astrocyte polarization. Rb1 has potential value for treating neurodegenerative diseases.     

Influence of moxibustion on NR2B and PKMζ after neural stem cell transplantation in the rats with vascular dementia: 艾灸对血管性痴呆大鼠神经干细胞移植后海马NR2B和PKMζ的影响

ObjectiveTo observe the influence of moxibustion on learning and memory ability in the rats with vascular dementia (VD) and explore the potential effect mechanism.MethodsA total of 80 rats, screened by Morris water maze, were randomly divided into a sham-operation group, a model group, a neural stem cells (NSCs) group, a NSCs + piracetam group and a NSCs + moxibustion group, 16 rats in each group. After corresponding treatments, Morris water maze and immunofluorescence technique were adopted to evaluate the therapeutic effect respectively.ResultsComparison among groups after modeling: compared with the sham-operation group, the escape latency was longer (P < 0.01) and the times of crossing platform were reduced (P < 0.01) in the rats of the model group. Comparison among groups after treatment: compared with the model group, the escape latency was shortened (P < 0.01) and the times of crossing platform were increased (P < 0.05) in the rats of the NSCs group. Compared with the NSCs group, the escape latency was shorter and the expressions of the hippocampus NR2B/EGFP and PKMζ/EGFP expression level increased (all P < 0.05) in the rats of the NSCs + piracetam group and the NSCs + moxibustion group (all P < 0.05). Compared with the NSCs + piracetam group, the escape latency was shorter and the expressions of the hippocampus NR2B/EGFP and PKMζ/EGFP expression level were higher in the rats of the moxibustion + NSCs group (all P < 0.05)ConclusionMoxibustion improves the spatial learning and memory ability of the VD rats and promotes the reconstruction of neurogenesis and synaptic function, which may be related to the up-regulation of the expressions of hippocampus NR2B and PKMζ expressions.     

Effects of Fuhe decoction on behaviors and monoamine neurotransmitters in different brain regions of CUMS combined with social isolation depression model rats

ObjectiveTo investigate the effect of Fuhe decoction on the behavior and levels of monoamine neurotransmitters in different brain regions in a depression rat model induced by chronic unpredictable mild stimulation (CUMS) combined with social isolation.MethodsFifty male SD rats were randomly divided into a blank group, model group, fluoxetine group, Chaiqinwendan decoction group, and Fuhe decoction group. Chronic unpredictable mild stimulation combined with a social isolation method was used to replicate the depression rat model. After 42 days of administration, a tail suspension test and high-performance liquid electrochemical detection (HPLC-ECD) were used to detect the behavioral changes and changes in the content of monoamine neurotransmitters norepinephrine (NE), dopamine (DA), 5-hydroxytrytamine (5-HT), and metabolites in different brain regions of rats in each group before and after treatment.ResultsCompared with the model group, the epinephrine (E) content in the Fuhe decoction group was highly significantly increased (P < .01). Compared with the model group, the 5-HT content of the prefrontal cortex in rats in the Fuhe decoction group was highly significantly increased (P < .01). Furthermore, compared with the model group, the 5-HT content in the hippocampus of rats in the Fuhe decoction group was significantly increased (P < .05).ConclusionFuhe decoction can improve the depression-like behaviors of model rats, and its antidepressant effect may be related to the increase in 5-HT content in the prefrontal cortex and hippocampus of rats.     

Correlation of gut microbiota and neurotransmitters in a rat model of post-traumatic stress disorder

ObjectiveTo determine the effect of gut microbiota on a rat model of post-traumatic stress disorder (PTSD) and explore the correlation of gut microbiota with behavior and neurotransmitters.MethodsWe established a single prolonged stress (SPS) model to examine the pathogenesis of PTSD on rat behavior, gut microbiota, and neurotransmitter levels. Rats were separated into control and model groups, and neurotransmitter levels were measured using enzyme linked immunosorbent assay. Then, 16 S rRNA sequencing was used to compare the gut microbiota between the control and model groups.ResultsCompared with those in the control group, freezing time significantly increased, while number of standing upright, crossing frequency, time spent in the central arena, and total distance traveled were significantly reduced in the model group after exposure to SPS (all P < .05). Meanwhile, serotonin, or 5-hydroxytryptamine, levels in the brain in the model group were significantly lower than those the control group (P = .0332). In addition, changes were observed in the gut microbiota diversity and relative abundances of bacterial phyla, orders, families, and genera in the model group. Especially, changes in Firmicutes, Bacteroidetes, Cyanobacteria, and Proteobacteria levels were most pronounced after SPS exposure. Correlation analysis showed that the strongest positive correlation was found between Bacteroidaceae and 5-HT (P = .0009). Moreover, RF32 abundance was the most negatively related to 5-HT (P = .0009), crossing frequency (P = .0007), and total distance (P = .0003).ConclusionOur results suggest that SPS model rats showed differences in behavior, neurotransmitter levels, and gut microbiota with control rats. Moreover, Firmicutes, Bacteroidetes, Cyanobacteria, and Proteobacteria were most relevant to the exhibited fear-like and anxiety-like behaviors and significant serotonin content reduction in SPS model rats.     

Manipulation for degenerative lumbar spondylolisthesis: A systematic review of randomized controlled trials

ObjectiveTo assess the effectiveness and safety of manipulation intervention for degenerative lumbar spondylolisthesis (DLS).MethodsThis is a systematic review and meta-analysis. A full-scale retrieval method was performed until February 1, 2021, including nine databases. The homogeneity of different studies was summarized using the Review Manager. The quality of studies was determined with the Cochrane risk-of-bias tool. The evidence quality was graded with the Grading of Recommendations, Assessment, Development, and Evaluations approach.ResultsA total of 6 studies involving 524 participants were included. The review demonstrated that manipulation has statistically significant improvements for treating DLS according to Japanese Orthopedic Association scores (mean difference, 3.76; 95% confidence interval, 2.63 to 4.90; P < .001) and visual analog scale scores (mean difference, ?1.50; 95% confidence interval, ?1.66 to ?1.33; P < .001) compared to the control group. One study reported that the difference in the Oswestry Disability Index between the traction group and the combination of manipulation and traction group was statistically significant (P < .05), while another reported that manipulation treatment can significantly improve the lumbar spine rotation angle on X-ray images compared with the baseline data (P < .05). Moreover, the manipulation group (experimental group) had fewer adverse events than the lumbar traction group (control group).ConclusionManipulation intervention is more effective and safer for DLS. Nevertheless, large-scale randomized controlled trials are required to confirm the current conclusions.