[Objective]To evaluate the effect of polysaccharide from Polygonatum sibiricum on Toll-like receptor 4(TLR4)activation and clarify its antigen specific response-enhancing effect on mice.[Methods]The HEK-BlueTM hTLR4 cell line which was stably transfected with genes of human TLR4 and secreted embryonic alkaline phosphatase(SEAP)was used to quickly evaluate the activation of four different types of polysaccharide from Polygonatum sibiricum,including Polygonatum kingianum NP H2O(PKNH),Polygonatum kingianum AP H2O(PKAH),Polygonatum kingianum NP 5%EtOH(PKNEt)and Polygonatum kingianum AP 5%EtOH(PKAEt)on TLR4.C57BL/10ScNJ(TLR4-/-)mice were used to determine the relationship between the drug effect and TLR4.Ovalbumin(OVA)immunized mouse model was used to evaluate the enhancement of drug on specific immune response.The serum immunoglobulin G1(IgG1)and the cytokines related to the TLR4 signaling pathways such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interferon-γ inducible protein-10(IP-10)levels were detected by enzyme-linked immunosorbent assay(ELISA),and lymphocyte proliferation was evaluated by 5-[3-(Aarboxymethoxy)phenyl]-3-(4,5-dimethyl-2-thiazolyl)-2-(4-sulfophenyl)-2H-tetrazolium innersalt(MTS)assay.The mRNA expression levels of cytokines related to the TLR4 signaling pathways was determined by reverse transcription-polymerase chain reaction(RT-PCR).[Results]TLR4 was activated most strongly by treatment of neutral polysaccharides from PKNH for 24 h(P<0.001).Compared with wild type,the cell proliferation and the expressions of TNF-α,IL-6 and IP-10 were significantly decreased in PKNH treated splenic lymphocyte from C57BL/10 ScNJ mice(P<0.001).Intragastric administration of PKNH significantly increased serum IgG1 level(P<0.01)and promoted splenic lymphocyte proliferation in OVA immunized mice(P<0.05).[Conclusion]The polysaccharide from Polygonatum sibiricum PKNH enhances the antigen specific immune response through activation of TLR4.

[Objective]To evaluate the effect of polysaccharide from Polygonatum sibiricum on Toll-like receptor 4(TLR4)activation and clarify its antigen specific response-enhancing effect on mice.[Methods]The HEK-BlueTM hTLR4 cell line which was stably transfected with genes of human TLR4 and secreted embryonic alkaline phosphatase(SEAP)was used to quickly evaluate the activation of four different types of polysaccharide from Polygonatum sibiricum,including Polygonatum kingianum NP H2O(PKNH),Polygonatum kingianum AP H2O(PKAH),Polygonatum kingianum NP 5%EtOH(PKNEt)and Polygonatum kingianum AP 5%EtOH(PKAEt)on TLR4.C57BL/10ScNJ(TLR4-/-)mice were used to determine the relationship between the drug effect and TLR4.Ovalbumin(OVA)immunized mouse model was used to evaluate the enhancement of drug on specific immune response.The serum immunoglobulin G1(IgG1)and the cytokines related to the TLR4 signaling pathways such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interferon-γ inducible protein-10(IP-10)levels were detected by enzyme-linked immunosorbent assay(ELISA),and lymphocyte proliferation was evaluated by 5-[3-(Aarboxymethoxy)phenyl]-3-(4,5-dimethyl-2-thiazolyl)-2-(4-sulfophenyl)-2H-tetrazolium innersalt(MTS)assay.The mRNA expression levels of cytokines related to the TLR4 signaling pathways was determined by reverse transcription-polymerase chain reaction(RT-PCR).[Results]TLR4 was activated most strongly by treatment of neutral polysaccharides from PKNH for 24 h(P<0.001).Compared with wild type,the cell proliferation and the expressions of TNF-α,IL-6 and IP-10 were significantly decreased in PKNH treated splenic lymphocyte from C57BL/10 ScNJ mice(P<0.001).Intragastric administration of PKNH significantly increased serum IgG1 level(P<0.01)and promoted splenic lymphocyte proliferation in OVA immunized mice(P<0.05).[Conclusion]The polysaccharide from Polygonatum sibiricum PKNH enhances the antigen specific immune response through activation of TLR4.

Objective There are few researches for the serum betatrophin level and diabetic nephropathy (DN) recently.The aim of this study was to investigate the change of serum betatrophin level and the correlation of serum betatrophin and urinary albumin-to-creatintine ratio (UACR) in patients with type 2 diabetes mellitus (T2DM).Methods A total of 150 Chinese subjects from Mar 2013 to Jul 2016 were enrolled in the study, including 90 patients with type 2 diabetes and 60 healthy controls.According to the level of UACR, the diabetic patients were divided into two groups:normal UACR group (UACR<30 mg/g, n=60) and abnormal UACR group(UACR>30 mg/g, n=30).Serum betatrophin was measured by enzyme linked immunosorbent assay (ELISA).UACR was measured by turbidimetric inhibition immune assay.Blood glucose blood lipid were measured simultaneously.Results The serum betatrophin level was significantly higher in abnormal UACR group than that in normal UACR group[677.37±59.02 vs 486.13±41.22 pg/mL, P<0.05];Serum betatrophin level in T2DM patients was positively correlated with age (r=0.246), waist hip ratio (WHR) (r=0.240), fasting blood glucose (FPG) (r=0.234), 2 hour plasma glucose (2hPG) (r=0.363), glycosylated hemoglobin (HbA1c) (r=0.346), fasting insulin (FINS) (r=0.249), insulin resistance index (HOMA-IR) (r=0.309), blood urea nitrogen (BUN) (r=0.223), creatinine (CREA) (r=0.277) and UACR (r=0.244) (P<0.05),and negatively correlated with glomerular filtration rate (GFR) (r=0.308) (P<0.01).Serum betatrophin level in normal UACR group was positively correlated with age, HbA1c and UACR (P<0.05);Serum betatrophin level in abnormal UACR group was positively correlated with WHR (r=0.504), 2hPG (r=0.600), HbA1c (r=0.449), HOMA-IR (r=0.395) (P<0.05).The WHR, HbA1c, HOMA-IR and GFR were the influential factors of the serum betatrophin level.Conclusion The level of serum betatrophin was significantly increased in T2DM patients with albuminuria, which suggests that the betatrophin might play an important role in the pathogenesis of DN.