Ramulus Cinnamomi （RC）, a traditional Chinese herb, has been used to attenuate inflammatory responses. The purpose of this study was to investigate the effect of RC extract on lipopolysaccharide （LPS）-induced neur...Ramulus Cinnamomi （RC）, a traditional Chinese herb, has been used to attenuate inflammatory responses. The purpose of this study was to investigate the effect of RC extract on lipopolysaccharide （LPS）-induced neuroinflammation in BV2 microglial cells and the underlying mechanisms involved. BV2 cells were incubated with normal medium （control group）, LPS, LPS plus 30 pg/mL RC extract, or LPS plus 100 pg/mL RC extract. The BV2 cell morphology was observed under an optical microscope and cell viability was detected by MTT assay. Nitric oxide level in BV2 cells was detected using Griess regents, and the levels of interleukin-6, interleukin-1 β, and tumor necrosis factor u in BV2 cells were determined by ELISA. The expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 proteins were detected by western blot assay. Compared with the LPS group, both 30 and 100 μg/mL RC extract had no significant effect on the viability of BV2 cells. The levels of nitric oxide, interleukin-6, interleukin-1β and tumor necrosis factor ct in BV2 cells were all significantly increased after LPS induction, and the levels were significantly reversed after treatment with 30 and 100 μg/mL RC extract. Furthermore, RC extract significantly inhibited the protein expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 in LPS-induced BV2 cells. Our findings suggest that RC extract alleviates neuroinflammation by downregulating the TLR4/MyD88 signaling pathway.展开更多
基金supported by a grant from the National Natural Science Foundation of China,No.81473383a grant from the Medical and Health Innovation Project of Chinese Academy of Medical Sciences,No.2016-I2M-3-007a grant from Key Project of New-Drugs Creation of Science and Technology of China,No.2012ZX09103101-078 and 2017ZX09101003-003-019
文摘Ramulus Cinnamomi （RC）, a traditional Chinese herb, has been used to attenuate inflammatory responses. The purpose of this study was to investigate the effect of RC extract on lipopolysaccharide （LPS）-induced neuroinflammation in BV2 microglial cells and the underlying mechanisms involved. BV2 cells were incubated with normal medium （control group）, LPS, LPS plus 30 pg/mL RC extract, or LPS plus 100 pg/mL RC extract. The BV2 cell morphology was observed under an optical microscope and cell viability was detected by MTT assay. Nitric oxide level in BV2 cells was detected using Griess regents, and the levels of interleukin-6, interleukin-1 β, and tumor necrosis factor u in BV2 cells were determined by ELISA. The expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 proteins were detected by western blot assay. Compared with the LPS group, both 30 and 100 μg/mL RC extract had no significant effect on the viability of BV2 cells. The levels of nitric oxide, interleukin-6, interleukin-1β and tumor necrosis factor ct in BV2 cells were all significantly increased after LPS induction, and the levels were significantly reversed after treatment with 30 and 100 μg/mL RC extract. Furthermore, RC extract significantly inhibited the protein expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 in LPS-induced BV2 cells. Our findings suggest that RC extract alleviates neuroinflammation by downregulating the TLR4/MyD88 signaling pathway.
文摘目的:探究右美托咪定(dexmedetomidine,DEX)对博莱霉素(bleomycin,BLM)诱导的肺纤维化小鼠的保护作用以及对Toll样受体4/髓样分化因子(toll-like receptor 4/myeloid differentiation factor88,TLR4/MyD88)信号通路的影响。方法:通过注射BLM诱导实验性小鼠肺纤维化。将小鼠分为生理盐水(Sham)组、BLM组、BLM+DEX组、BLM+PFD组和BLM+DEX+LPS组。苏木素-伊红(hematoxylin and eosin,HE)染色和Masson染色分析肺组织病理学变化;免疫组织化学检测纤连蛋白(fibronectin,Fn)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)和Ⅰ型胶原蛋白(collagenⅠ)表达情况;原位缺口末端标记法(TdT mediated d UTP nick end labeling,TUNEL)检测肺组织中细胞凋亡;检测支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中总细胞数量及蛋白质含量;Western blot检测肺组织中TLR4/MyD88信号通路和细胞凋亡相关蛋白水平。结果:与Sham组相比,BLM刺激后肺组织中肺泡炎评分[(2.56±0.24)分vs.(0.15±0.02)分]和Ashcroft评分[(5.68±0.52)分vs.(0.09±0.01)分]明显增高,Fn[(63.63±5.48)%vs.(25.12±2.16)%]、α-SMA[(58.63±5.03)%vs.(17.56±1.25)%]和collagenⅠ[(55.32±5.16)%vs.(12.03±1.20)%]表达升高(P<0.05),肺部炎症程度和纤维化程度增加;同时TUNEL阳性细胞率增高,细胞中bcl-2相关X蛋白(bcl-2 associated X protein,Bax)水平增高,B淋巴细胞瘤-2蛋白(B-cell lymphoma-2 protein,Bcl-2)水平降低(P<0.05);BALF中总细胞数量[(3.54±0.06)×10^(5)个/mL vs.(0.98±0.07)×10^(5)个/mL]和蛋白质含量[(2.85±0.20)mg/mL vs.(0.29±0.03)mg/mL]升高(P<0.05);肺组织中TLR4和MyD88蛋白水平升高(P<0.05)。DEX治疗可减轻BLM诱导的小鼠肺纤维化和炎症,降低TUNEL阳性细胞率,并逆转Fn、α-SMA、collagenⅠ、Bax和Bcl-2表达(P<0.05);同时DEX也降低BLM诱导的小鼠BALF中总细胞数量和蛋白质含量(P<0.05)。此外,DEX降低BLM诱导的小鼠肺组织中TLR4和MyD88蛋白水平(P<0.05)。吡非尼酮(pirfenidone,PFD)与DEX具有相似
