Longdanxiegan granules resistant genital herpes virus and on dendritic cell function in vitro

Objective: To study medicine Longdanxiegan Granules for genital herpes virus proliferation; through research Longdanxiegan Granules for genital herpes in patients with peripheral blood dendritic cell surface signs of maturity CD83, CD86 expression and effects on dendritic shaped cells secrete interleukin-12 levels, and explore Longdanxiegan granules immune mechanism of action targets. Method: 1) using cell culture technology for water soluble granules Longdanxiegan objects outside the efficacy of anti-HSV-2 experimental research. The proliferation inhibition assay, and directly kill infected occlusion method three kinds of mode of action, different concentrations of the drug for genital herpes virus proliferation. 2) by in vitro cell culture methods used Longdanxiegan Water soluble granules for genital herpes in patients with the role of dendritic cells, and then detects the peripheral blood of patients and healthy DC surface markers CD83, CD86 expression levels. 3) detected by ELISA normal control group and patients with genital herpes group of DC culture supernatant IL-12 content. Results: 1) drug cytotoxicity assay: normal control cells were less than 25% of the CPE, Longdanxiegan granules and acyclovir (ACV) on Vero cells the maximum non-toxic concentration (TC0) were 0.16,0 . lmg / ml. calculated by Reed-Muench method Longdanxiegan Granules, ACV TC50 on Vero cells were 5.631,0.457 mg / ml. 2) Determination of viral infection: the virus stock line delivery times for 10-fold dilution for determination of viral infection, the measured HSV-2 333 strains TCID50 = 10-4/0.1ml. 3) drugs on the CPE caused by HSV-2 suppression results: three different medications to join mode, virus control group established by experiment are visible typical CPE, normal cells in the control group had no significant lesions. 4) MTT method Drug Efficacy of anti-HSV-2: normal cells was determined by MTT control group and the virus control group, the average A490 values ??were 0.294 ± 0.053,0.032 ± 0.0065. Mode Ⅰ dosing, the concentration of almost all experimental group and western medicine control group, the average A490 values ??with the virus control group were statistically significant P (lt; 0.05 or P lt; 0.01); way Ⅱ dosing, partial concentration of medicine A490 experimental group, the average value of the control group compared with the virus, a significant difference (P lt; 0.05); way Ⅲ dosing, almost all experimental groups, the average concentration of medicine A490 values ??with the virus control group were statistically significant ( P lt; 0.05 or P lt; 0.01); way Ⅱ and Ⅲ dosing, the A490 comparison group mean and virus control group, there was no statistically significant difference (P gt; 0.05). 5) to drug concentration as abscissa (μg / ml), viral suppression rate (IR%) plotted as ordinate manner different drugs added dose-response relationship diagram. Way IACV on the inhibition of HSV-2 was enhanced with the concentration there is an obvious dose-effect relationship; while the amount of particles Longdanxiegan effect relationship was not significant. Concentration of 40μg/ml medicine, its inhibitory effect on HSV-2 but decreased with increasing concentration. TCM mode Ⅱ concentration exceeds 20μg/ml, its inhibition of HSV-2 was enhanced with the concentration. Way Ⅲ medicine also occurs when the concentration of 80μg/ml concentration decreased with IR. 6) normal control group DC surface markers CD83, CD86 expression is low; while the normal control group TCM DC surface markers CD83, CD86 expression was significantly increased, compared with the control group, there were significant differences (P lt ; 0.05); genital herpes in patients in the control group of DC surface markers CD83, CD86 expression was significantly increased compared with normal control group were significantly different (P lt; 0.01). Genital herpes medicine medicine after action group to join DC surface markers CD83, CD86 and downregulated the control group of patients, there is a significant difference (P lt; 0.01) 7) normal control group of IL-12 content of 84.57ng / ml, genital herpes in patients with normal control group, the content of the blank control group was significantly higher, there is a significant difference (P lt; 0.01); while in the control group of normal Chinese culture supernatant of IL-12 content and normal control group compared decreased, but the difference was not statistically significant (P gt; 0.05). Medicines in patients with genital herpes culture supernatant levels of IL-12 compared with the control group of patients was significantly reduced, there is a significant difference (P lt; 0.01). Conclusions: 1) concentration of less than 0.16mg/ml of Longdanxiegan Granules on Vero cells non-toxic, very safe on Vero cells. 2) Longdanxiegan granules and acyclovir could inhibited to varying degrees within the HSV-2 Vero cell proliferation Longdanxiegan Granules for HSV-2 has a direct role in the killing, and for HSV-2 in infection have some preventive effect, and ACV no direct killing and prevention. Longdanxiegan granules in vitro effects from multiple strong links to suppress HSV-2. 3) ACV against HSV-2 inhibition of proliferation in a dose-response relationship; Longdanxiegan Granules for HSV-2 infection blocking role of dose-effect relationship exists; but its effect on the proliferation of HSV-2 inhibition and direct killing for no apparent dose-response relationship. 4) Chinese medicine granules Longdanxiegan effect on normal human peripheral blood dendritic cells, antigen presenting cells significantly enhanced the ability of Chinese medicine acting on genital herpes in patients with peripheral blood dendritic cells, antigen presenting cells of some inhibitory ability, indicating Longdanxiegan granules on peripheral blood dendritic cell antigen-presenting function play a regulatory role. 5) Chinese medicine on dendritic cells in both normal subjects and patients, the dendritic cell cytokine production of IL-12 strongly inhibited, and far lower than the normal level, indicating Longdanxiegan mainly in the liver granules cytokine IL-12 production is to suppress the point of differentiation of Th1 lymphocytes to treat genital herpes.