Antiviral effect of Chinese herbal prescription JieZe-1 on adhesion and penetration of VK2/E6E7 with herpes simplex viruses type 2

Abstract

Ethnopharmacological relevance: The Chinese Herbal Prescription JieZe-1(JZ-1), added and subtracted from Yihuang Decoction, a famous formula in the 12th year of Kangxi in Qing Dynasty, has a clear effect on Genital Herpes (GH) and no obvious adverse reactions occur clinically. JZ-1 also has preventive and therapeutic effects on Trichomonas vaginitis, Candida albicans vaginitis and GH in vitro and in vivo experiments. Aim of study: The effect and mechanism of JZ-1 on anti-herpes simplex virus type 2(HSV-2) in vitro focusing on adhesion and penetration stages were investigated. Materials and methods: A model of HSV-2 infection of VK2/E6E7 was developed. In order to explore JZ-1's anti-HSV-2 effect in vitro, cell morphology, ultrastructural pathology, cell viability and expression of viral glycoprotein D (gD) were assessed at 6 h, 12 h, 18 h, and 24 h of JZ-1 treatment. Then we measured the exact time required for adhesion and penetration of HSV-2 into VK2/E6E7 among a series of times at room temperature and under temperature control techniques. We treated VK2/E6E7 with JZ-1, penciclovir, or berberine and explored the mechanism of JZ-1 in blocking HSV-2 adhesion and penetration of host cells by assessing the cell ultrastructural pathology, viability, viral proteins gB, gD, VP16, ICP5, and ICP4 and host cell proteins HVEM, Nectin-1, and Nectin-2. Results: HSV-2 can fully adhere and penetrate into VK/E6E7 within 5 mins at room temperature while it takes 60mins under temperature control techniques. JZ-1 and penciclovir showed significant anti-HSV-2 effects, with improved host cell morphologies and increased host cell viabilities observed after treatment for 24 h. The anti-HSV-2 effect of JZ-1 can be detected after treatment for 6 h while that of penciclovir was not obvious until treatment for 12 h. JZ-1 showed distinct effect on HSV-2 adhesion and penetration stages by significantly reducing the expression of viral proteins gB, gD, VP16, ICP5, and ICP4, improving cell morphology and increasing cell viability. However, these effects were not exerted via downregulated expression of membrane fusion-related proteins such as HVEM, Nectin-1, or Nectin-2. The specific anti-HSV-2 mechanism of JZ-1 need to be further explored. Conclusion: The anti-HSV-2 effect of JZ-1 was superior to that of penciclovir and berberine in vitro, and was mainly mediated by enhancing host cell defense and blocking adhesion and penetration of HSV-2.

The Chinese herbal prescription JZ-1 induces autophagy to protect against herpes simplex Virus-2 in human vaginal epithelial cells by inhibiting the PI3K/Akt/mTOR pathway

“…Because of the complex components in the JZ-1 extract, HPLC fingerprinting was used to control the chemical components in the extract better. The results of HPLC fingerprint have been shown in a previous article (Duan et al, 2019), a total of six JZ-1 components were determined, namely berberine, paeoniflorin, neoastilbin, astilbin, paeonol and dictamnine.…”

“…JZ-1 consists of 10 traditional Chinese medicines, and the details of the herbal ingredients are shown in Table 1. JZ-1 was prepared as indicated previously (Duan et al, 2019). Briefly, Phellodendri Chinensis Cortex was extracted twice and filtered.…”

“…The primary chemical constituents of JZ-1 were identified by 3Dhigh performance liquid chromatography (3D-HPLC) method and were described in detail in a previous article (Duan et al, 2019).…”

“…Cytotoxicity and anti-HSV-2 activity of JZ-1 were examined by MTT assays following the method described in a previous article (Duan et al, 2019). Briefly, exponentially growing VK2/E6E7 cells were seeded into 96-well culture plates for 24 h, then various concentrations (0.625, 1.25, 2.5, 5, 10 and 20 mg/mL) of JZ-1 were added to the VK2/E6E7 cells for another 24 h in the presence or absence of HSV-2.…”

“…Our previous experiments showed that JZ-1 decoction could protect human vaginal epithelial cells against HSV-2. The anti-HSV-2 effect of JZ-1 is superior to that of berberine or penciclovir, and the effect is mainly achieved by augmenting the host cell defenses ability and blocking the adhesion and penetration of HSV-2 (Duan et al, 2019). In a model of HSV-2-infected VK2/E6E7 cells, the expression of the HSV-2 envelope protein gD increased gradually over time, and JZ-1 reduced this gD expression but had no effect on HVEM in the human vaginal epithelial cells infected by HSV-2 (Ai and Chen, 2017;Qiao and Chen, 2016).…”