1.Study on Intervention Effect of Sang-Ji Mixture in TLR-4 Expression on Peripheral Blood Monocytes in Hypertensive Patients with Overabundant Liver-fire and Phlegm Syndrome
Keke ZUO ; Mingjun ZHANG ; Ning GU
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(5):1103-1107
This study was aimed to observe the effect of Sang-Ji (SJ) mixture in the treatment of Toll-like receptor 4 (TLR-4) expression on peripheral blood monocytes (PBMCs) in hypertensive patients. A total of 60 cases of hyper-tensive cases with overabundant liver-fire and phlegm syndrome were randomly divided into the treatment group and the control group with 30 cases in each group. Both groups received Felodipine and/or Benazapril treat-ment. The treatment group was added with SJ mixture. The observation duration was 28 days. The detection was made on indicators such as blood pressure , traditional Chinese medicine ( TCM ) syndrome scores and TLR-4 ex-pression on PBMCs before and after treatment. The results showed that after treatment, the blood pressure, TCM syndrome scores and TLR-4 expression on PBMCs of both groups decreased compared with that of the pretreat-ment . Changes on TCM syndrome scores and TLR-4 expression of the treatment group were more significant than that of the control group ( P < 0 . 05 ) . It was concluded that SJ mixture improved clinical symptoms in pa-tients with overabundant liver-fire and phlegm syndrome , decreased TLR-4 expression on PBMCs , in order to inhibit the immune response to a certain extent .
2.Effects of ligustilide on the extracellular recombinant human heat shock protein 60 induced inflammatory reactions in the THP-1 cells and the related mechanisms
Ying GAO ; Rui CHEN ; Ning GU ; Xiaoli HE ; Zhijun YANG ; Keke ZUO
Chinese Journal of Cardiology 2016;44(9):793-798
Objective To investigate the effects of ligustilide (LIG) on extracellular recombinant human heat shock protein 60 (HSP60) induced inflammatory reactions in the THP-1 cells and the related mechanisms.Methods THP-1 cells were differentiated to macrophages by incubation with phorbol-12-myristate-13-acetate (PMA).The immunofluorescence method was used to screen the optimum transfection concentration of MyD88 siRNA.The macrophages were divided into six groups (n =3),including blank control (siRNA transfection reagent),model (siRNA transfection reagent + HSP60 10 mg/L),negative control (MyD88 negative control + HSP60 10 mg/L),LIG group(siRNA transfection reagent + HSP60 10 mg/L +LIG 20 mg/L),RNA interfering(RNAi) group(MyD88 siRNA + HSP60 10 mg/L) and RNAi + LIG group (MyD88 siRNA + HSP60 10 mg/L + LIG 20 mg/L).The protein expression level of MyD88 and phosphonuclear factor-κB (p-NF-κB) in macrophages and the level of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the culture supernatant were assessed by Western blot analyses or ELISA,respectively.Results (1)The protein expression levels of MyD88 (1.196 ± 0.125 vs.0.341 ± 0.063,P < 0.01) and p-NF-κB(0.817 ±0.034 vs.0.312 ±0.046,P <0.01) were significantly higher in the model group than those in the blank control group.The protein expression levels of MyD88 (0.554 ± 0.043) and p-NF-κB(0.538 ±0.063) in the RNAi group were significantly lower than those in the model group (all P <0.01) but significantly higher than those in the blank control group (all P < 0.05).The protein expression levels of MyD88(0.694 ±0.087,P <0.05) and p-NF-κB(0.669 ±0.043,P <0.01)in the LIG group were markedly lower than those in the model group,but higher than those in the RNAi group (P < 0.05) and the blank control group (P < 0.01).The protein expression levels of MyD88 (0.409 ± 0.069) and p-NF-κB(0.395 ±0.046) in the RNAi + LIG group were significantly lower than in the model group (all P < 0.01) and in the LIG group (P < 0.05 or 0.01),and were similar to the blank control group (P > 0.05).The expression level of p-NF-κB in the RNAi + LIG group was significantly lower than in the RNAi group (P<0.05).(2) The contents ofTNF-α((312.24±28.69) ng/Lvs.(5.99 ±1.03) ng/L,P< 0.01) and IL-6 ((233.45 ± 57.77) ng/L vs.(2.25 ± 0.67) ng/L,P < 0.01) were significantly higher in the model group than in the blank control group.The contents of TNF-α((235.66 ±25.12) ng/L) and IL-6((131.59 ± 13.99) ng/L) were significantly lower in the RNAi group than in the model group (P <0.01) The contents of TNF-α ((258.13 ± 44.80) ng/L) and IL-6 ((175.92 ± 28.27) ng/L) were also significantly lower in the LIG group than in the model group (P < 0.05) while the content of IL-6 was significantly higher in the LIG group than in the RNAi group(P <0.01).The contents of TNF-α((88.57 ± 16.10) ng/L) and IL-6((59.99 ± 10.31) ng/L) were significantly lower in the RNAi + LIG group than those in the model group,the RNAi group and the LIG group (P < 0.05 or 0.01).Conclusions The MyD88/NF-κB signaling pathway is one of the key signaling pathways of human HSP60 induced inflammation in THP-1 cells.Ligustilide could exhibit the anti-inflammatory effect probably by inhibiting the MyD88/NF-κB signaling pathway.