ABSTRACT
ObjectiveTo study the inhibitory effect of Banxia Houputang (BHT) on lipopolysaccharide (LPS)-induced inflammation of microglia (BV2) cells and the neuroprotective effect on human neuroblastoma (SH-SY5Y) cells. MethodAfter the neuroinflammatory model was constructed by LPS inducing BV2 cells, model group (LPS 100 µg·L-1), administration groups (LPS+1 g·L-1 BHT, LPS+2 g·L-1 BHT, LPS+5 g·L-1 BHT, LPS+10 g·L-1 BHT), and blank group were given DEME medium at the same volume. In addition, neuronal apoptosis model was established by co-culture of LPS-induced BV2 cell inflammation medium and SH-SY5Y cells (LPS-DMEM) and was administrated according to the above grouping. Cell viability was detected by Cell Counting Kit-8 (CCK-8) assay. The content of nitric oxide (NO) and that of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) were determined by Griess aasay and enzyme-linked immunosorbent assay (ELISA), respectively. The mRNA levels of TNF-α, IL-1β, interleukin-4 (IL-4), nitric oxide synthase (iNOS), and interleukin-10 (IL-10) were measured by real-time polymerase chain reaction (Real-rime PCR). Western blot was used to detect the expression levels of signal transducer and activator of transcription 3 (STAT3), Janus kinase 2 (JAK2) and nuclear factor kappa-B (NF-κB p65), protein kinase B (Akt), inhibitor of nuclear factor κB α (IκBα), B-cell lymphoma-2 (Bcl-2), and Bcl-2 associated X protein (Bax). ResultCompared with blank group, LPS increased the NO release, levels of TNF-α, IL-1β, IL-6, and iNOS and protein expression of Akt, NF-κB p65, IκBα, JAK2 and STAT3, decreased the content of IL-4 and IL-10 in BV2 cells, and induced apoptosis of co-cultured SH-SY5Y cells (P<0.01). Compared with model group, BHT reduced the content of NO, TNF-α, IL-1β, and iNOS (P<0.01) and protein expression of Akt, NF-κB p65, IκBα, JAK2 and STAT3 (P<0.01), elevated the content of IL-4 and IL-10 (P<0.01), and inhibited the apoptosis of SH-SY5Y cells induced by LPS-DMEM (P<0.01). ConclusionThis experiment reveals that BHT inhibited LPS-induced inflammation in BV2 cells by regulating Akt/NF-κB/JAK2/STAT3 signaling pathway and showed neuroprotective effects on SH-SY5Y cells.
ABSTRACT
ObjectiveTo reveal the mechanism of action of Huangqi Guizhi Wuwutang in the treatment of rheumatoid arthritis by pharmacological research based on its clinical application. MethodThe collagen-induced arthritis (CIA) rat model was established by injecting bovine type Ⅱ collagen and Freund's adjuvant at the tail, and was treated with different concentrations of Huangqi Guizhi Wuwutang. The rats were randomly divided into blank group, model group, methotrexate (0.9 mg·kg-1) group, and Huangqi Guizhi Wuwutang low- and high-dose (5.13, 20.52 g·kg-1·d-1) groups, with continuous intragastric administration for 4 weeks. The degree of joint swelling, weight, degree of foot swelling and arthritis index score were determined and the pathological changes of ankle joints were detected by hematoxylin and eosin (HE) staining to observe the therapeutic effect of Huangqi Guizhi Wuwutang on rheumatoid arthritis. In addition, enzyme-linked immunosorbent assay (ELISA) and Western blot were used to measure the expression of interleukin 1β (IL-1β), interleukin 6 (IL-6), interleukin 10 (IL-10) and tumor necrosis factor-α (TNF-α) in serum and the expression of nuclear factor kappa-B (NF-κB) pathway related proteins in synovial tissue, respectively to clarify the molecular mechanism of Huangqi Guizhi Wuwutang in the treatment of rheumatoid arthritis. ResultCompared with the conditions in blank group, the body weight and IL-10 level were decreased (P<0.01), and the degree of foot swelling and arthritis index score, the levels of IL-1β, IL-6 and TNF-α, and the expression of NF-κB pathway related proteins were increased (P<0.01,) in the model group, with impaired morphology and function of the ankle joint. Additionally, compared with the model group, Huangqi Guizhi Wuwutang low- and high-dose groups had increased body weight of rats and IL-10 level (P<0.01), and reduced degree of foot swelling and arthritis index score (P<0.05, P<0.01), levels of IL-1β, IL-6 and TNF-α (P<0.01) and expression of NF-κB pathway related proteins (P<0.05, P<0.01), with improved function and morphology of the ankle joint. ConclusionHuangqi Guizhi Wuwutang can significantly alleviate joint inflammatory injury by down-regulating NF-κB pathway and reducing the inflammatory response in CIA rats.
ABSTRACT
In this paper, the extraction rate of crude polysaccharides and the yield of polysaccharides from Hippocampus served as test indicators. The comprehensive evaluation indicators were assigned by the R language combined with the entropy weight method. The Box-Behnken design-response surface methodology(BBD-RSM) and the deep neural network(DNN) were employed to screen the optimal parameters for the polysaccharide extraction from Hippocampus. These two modeling methods were compared and verified experimentally for the process optimization. This study provides a reference for the industrialization of effective component extraction from Chinese medicinals and achieves the effective combination of modern technology and traditional Chinese medicine.