[Aucubin combined with ADSCs-exos protects TBHP-induced nucleus pulposus cells via TLR4/NF-κB pathway].

This paper aims to investigate the effects and mechanisms of adipose-derived stem cells-exosomes(ADSCs-exos) toge-ther with aucubin in protecting human-derived nucleus pulposus cells(NPCs) from inflammatory injury, senescence, and apoptosis. The tert-butyl hydroperoxide(TBHP)-induced NPCs were assigned into normal, model, aucubin, ADSCs-exos, and aucubin+ADSCs-exos groups. The cell viability was examined by cell counting kit-8(CCK-8), cell proliferation by EdU staining, cell senescence by senescence-associated-ß-galactosidase(SA-ß-Gal), and cell cycle and apoptosis by flow cytometry. Enzyme-linked immunosorbent assay was employed to examine the expression of interleukin-1ß(IL-1ß), IL-10, and tumor necrosis factor-α(TNF-α). Real-time fluorescence quantitative PCR and Western blot were employed to determine the mRNA and protein levels of aggregated proteoglycan(aggrecan), type Ⅱ collagen alpha 1(COL2A1), Toll-like receptor 4(TLR4), and nuclear factor-kappa B(NF-κB). The results showed that compared with the model group, the aucubin or ADSCs-exos group showed enhanced viability and proliferation of NPCs, decreased proportion of G_0/G_1 phase cells, increased proportion of S phase cells, reduced apoptosis and proportion of cells in senescence, lowered IL-1ß and TNF-α levels, elevated IL-10 level, down-regulated mRNA and protein levels of TLR4 and NF-κB, and up-regulated mRNA and protein levels of aggrecan and COL2A1. Compared with the aucubin or ADSCs-exos group, the aucubin+ADSCs-exos combination further increased the viability and proliferation of NPCs, decreased the proportion of G_0/G_1 phase cells, increased the proportion of S phase cells, reduced the apoptosis and proportion of cells in senescence, lowered the IL-1ß and TNF-α levels, elevated the IL-10 level, down-regulated the mRNA and protein levels of TLR4 and NF-κB, and up-regulated the mRNA and protein levels of aggrecan and COL2A1. In summary, both aucubin and ADSCs-exos could exert protective effects by inhibiting inflammatory responses, reducing apoptosis and senescence of NPCs, improving cell viability and proliferation as well as extracellular matrix synthesis, which may be associated with the inhibition of TLR4/NF-κB signaling pathway activation. The combination of both plays a synergistic role in the protective effects.

Comparative efficacy and safety of Chinese herbal medicine for knee osteoarthritis: A protocol for systematic review and network meta-analysis.

BACKGROUND: Knee osteoarthritis (OA) is a major public health concern causing chronic disability as well as a substantial burden on health care and the economy. However, effective treatments for knee OA were still not available. Numerous clinical studies have suggested that Chinese herbal medicine (CHM) seems to be clinically effective in treating knee OA. Thus, this study aims to evaluate the efficacy and safety of CHM in the treatment of knee OA through a systematic review and network meta-analysis. METHODS: A comprehensive search will be performed in PubMed, Cochrane Library, Embase, Web of Science, China National Knowledge Infrastructure, VIP Database, Wanfang Database, Chinese Biomedical Database, and 3 clinical trials registration websites, from the database inception to May 2021. Randomized controlled trials meeting the eligible criteria based on the PICOS framework will be included. All studies fulfilling the eligible criteria will be assessed for risk of bias using the Cochrane Collaboration's tool. The primary outcome will be the visual analog scale (VAS), Western Ontario and McMaster Universities Osteoarthritis Index, and total effective rate. The secondary outcome is the incidence of adverse events. Data analysis will be performed using Stata, Addis, and WinBUGS. DISCUSSION: This study will provide a reliable evidence to assess effectiveness and safety of CHM for knee OA, which may provide guidance for clinical practice. SYSTEMATIC REVIEW REGISTRATION: This study protocol has been registered on INPLASY202160060.

[Influence of Aloe polysaccharide on proliferation and hyaluronic acid and hydroxyproline secretion of human fibroblasts in vitro].

OBJECTIVE: To investigate the effect of Aloe polysaccharide on proliferation and hyaluronic acid and hydroxyproline secretion of human fibroblasts in vitro. METHODS: The fibroblasts were treated with different doses of polysaccharide (0, 25, 50, 100, 200, 400 mg/L). Subsequently, cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, cell cycle by flow cytometry, evaluation of the Aloe polysaccharide toxic effect by acridine orange-ethidium bromide staining, evaluation of the cell injury by lactate dehydrogenase (LDH) assay, and the collagen synthesis by (3)H-proline incorporation. In addition, hyaluronic acid and hydroxyproline levels in the supernatants of cultured fibroblasts were measured by enzyme-linked immunosorbent assay. RESULTS: The proliferation of fibroblasts was induced with polysaccharide in a dose-dependent manner, reaching its highest level on 5th day. Meanwhile, the percentage of cells at phase G(0)/G(1) was decreased, while that at phases G(2)/M and S was increased significantly in Aloe polysaccharide-treated groups as compared with those in the control group (P<0.05). Additionally, the apoptosis of the fibroblasts showed no differences among all groups. The collagen synthesis was increased and cell injury decreased in polysaccharide-treated groups as compared with those in control group (P<0.05), while the levels of hyaluronic acid and hydroxyproline in the supernatants of fibroblasts treated with polysaccharide were significantly higher than those in the control group (P<0.05). CONCLUSION: The Aloe polysaccharide promotes both the proliferation of fibroblasts and the production of hyaluronic acid and hydroxyproline in fibroblasts. This indicates that the Aloe polysaccharide may play an important role in the extracellular matrix remodeling during wound healing.

[The effects of aloe extract on nitric oxide and endothelin levels in deep-partial thickness burn wound tissue in rat].

OBJECTIVE: To investigate the effects of polysaccharide extracted from Aloe barbadensis and Aloe barbedensis containing gel on tissue water contents, nitric oxide (NO) and endothelin (ET) levels in wounds of burned rats. METHODS: Four areas of deep-partial thickness burn wounds with 3 cm in diameter were made on each back of 42 male Wistar rats. Single layer gauze impregnated either with 5% (W/W) aloe raw polysaccharide, 10% (W/W) aloe gel, 1% (W/W) sulfadiazine pyridine silver cream (SD-Ag), or normal saline was respectively applied on different wounds. According to different medications, the wounds were divided into aloe raw polysaccharide group, aloe gel group, SD-Ag group and normal saline group. Six rats in each group were sacrificed at 4, 12, 24, 48 post-scald hour (PSH) and on 7, 14, 21 post-scald day (PSD), and the full-thickness skin of wound was harvested for the determination of wound tissues water contents, NO and ET levels, and for calculation of NO/ET ratio. Another 6 normal rats served as normal controls. RESULTS: The water content in the wound tissue in aloe raw polysaccharide group at 12, 24 and 48 PSH [(73.4 +/- 3.8)%, (76.6+/-3.0)%, (70.6+/-3.8)%] and aloe gel group [(74.5+/-2.6)%, (77.1+/-3.6)%, (71.2 +/- 3.1)%] was obviously lower than those in SD-Ag group [(80.1 +/- 4.1)%, (80.5 +/-3.9)%, (76.1 +/-3.8)%, P <0.05]. During 7-21 PSD, all of them returned to the normal level except that in SD-Ag group, as it was still higher than that in normal controls (P < 0.05). The NO content in wound tissue in each group reached the peak at 12 PSH, decreased thereafter, but it was still obviously higher than that of normal controls on 21 PSD (P < 0.05). The ET content in wound tissue of each group reached the peak on 7 or 14 PSD, decreased thereafter, but it was still evidently higher than that in normal controls on 7 or 14 PSD (P < 0.05). The NO content in wound tissue in aloe raw polysaccharide and aloe gel group were markedly lower than those in SD-Ag and normal saline groups at 12 and 24 PSH ( P < 0.05). The NO/ET ratio in each group reached the peak at 12 PSH, decreased thereafter, and it returned to normal value on 14 PSD. On 7 PSD, the NO/ET ratio in aloe gel, SD-Ag and normal saline groups were still significantly higher than that in normal controls, except that returned to normal value in aloe raw polysaccharide group. CONCLUSION: Both aloe raw polysaccharide and aloe gel can decrease wound tissue NO release, optimize NO/ET ratio, lighten vascular inflammatory reaction, and lessen permeability and edema.

[Effect of Aloe coarse polysaccharide on cytokine secretion of keratinocytes in vitro].

OBJECTIVE: To study the effects of Aloe coarse polysaccharide on the levels of growth factors (EGF, TGF-alpha, TGF-beta1) and interleukins (IL-1beta, IL-6, IL-8) and tumor necrosis factor (TNF) in cultured keratinocytes. METHOD: The cultured keratinocytes were treated with Aloe coarse polysaccharide at concentrations of 75, 150, 300, 600, 1 200 mg x L(-1) land the equal volume of media as control group. The levels of EGF, TGF-alpha, TGF-beta1, IL-1beta, IL-6, IL-8 and TNF in the supernatants of cultured keratinocytes were assayed by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA). RESULT: Compared with the control group, the levels of EGF, TGF-alpha, IL-1beta, IL-6 and IL-8 were significantly increased by treatment with Aloe coarse polysaccharide (P < 0.05, P < 0.01) and in a dose dependent manner, and the levels of TGF-beta1 and TNF were also increased but no statistical significance. CONCLUSION: Aloe coarse polysaccharide may promote keratinocytes to secrete EGF, TGF-alpha, IL-1beta, IL-6 and IL-8.