OBJECTIVE: JieZe-1 (JZ-1), a Chinese herbal prescription, has an obvious effect on genital herpes, which is mainly caused by herpes simplex virus type 2 (HSV-2). Our study aimed to address whether HSV-2 induces pyroptosis of VK2/E6E7 cells and to investigate the anti-HSV-2 activity of JZ-1 and the effect of JZ-1 on caspase-1-dependent pyroptosis. METHODS: HSV-2-infected VK2/E6E7 cells and culture supernate were harvested at different time points after the infection. Cells were co-treated with HSV-2 and penciclovir (0.078125 mg/mL) or caspase-1 inhibitor VX-765 (24 h pretreatment with 100 μmol/L) or JZ-1 (0.078125-50 mg/mL). Cell counting kit-8 assay and viral load analysis were used to evaluate the antiviral activity of JZ-1. Inflammasome activation and pyroptosis of VK2/E6E7 cells were analyzed using microscopy, Hoechst 33342/propidium iodide staining, lactate dehydrogenase release assay, gene and protein expression, co-immunoprecipitation, immunofluorescence, and enzyme-linked immunosorbent assay. RESULTS: HSV-2 induced pyroptosis of VK2/E6E7 cells, with the most significant increase observed 24 h after the infection. JZ-1 effectively inhibited HSV-2 (the 50% inhibitory concentration = 1.709 mg/mL), with the 6.25 mg/mL dose showing the highest efficacy (95.76%). JZ-1 (6.25 mg/mL) suppressed pyroptosis of VK2/E6E7 cells. It downregulated the inflammasome activation and pyroptosis via inhibiting the expression of nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing protein 3 (P < 0.001) and interferon-γ-inducible protein 16 (P < 0.001), and their interactions with apoptosis-associated speck-like protein containing a caspase recruitment domain, and reducing cleaved caspase-1 p20 (P < 0.01), gasdermin D-N (P < 0.01), interleukin (IL)-1β (P < 0.001), and IL-18 levels (P < 0.001). CONCLUSION JZ-1 exerts an excellent anti-HSV-2 effect in VK2/E6E7 cells, and it inhibits caspase-1-dependent pyroptosis induced by HSV-2 infection. These data enrich our understanding of the pathologic basis of HSV-2 infection and provide experimental evidence for the anti-HSV-2 activity of JZ-1. Please cite this article as: Liu T, Shao QQ, Wang WJ, Liu TL, Jin XM, Xu LJ, Huang GY, Chen Z. The Chinese herbal prescription JieZe-1 inhibits caspase-1-dependent pyroptosis induced by herpes simplex virus-2 infection in vitro. J Integr Med. 2023; 21(3): 277-288.
Caspase 1/metabolism* ; Inflammasomes/pharmacology* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Pyroptosis ; Simplexvirus/metabolism* ; Drugs, Chinese Herbal/pharmacology* ; Herpes Simplex/drug therapy* ; Humans

The study investigated the inhibitory effect and mechanism of tectorigenin derivative(SGY) against herpes simplex virus type Ⅰ(HSV-1) by in vitro experiments. The cytotoxicity of SGY and positive drug acyclovir(ACV) on African green monkey kidney(Vero) cells and mouse microglia(BV-2) cells was detected by cell counting kit-8(CCK-8) method, and the maximum non-toxic concentration and median toxic concentration(TC_(50)) of the drugs were calculated. After Vero cells were infected with HSV-1, the virulence was determined by cytopathologic effects(CPE) to calculate viral titers. The inhibitory effect of the tested drugs on HSV-1-induced cytopathy in Vero cells was measured, and their modes of action were initially explored by virus adsorption, replication and inactivation. The effects of the drugs on viral load of BV-2 cells 24 h after HSV-1 infection and the Toll-like receptor(TLR) mRNA expression were detected by real-time fluorescence quantitative PCR(RT-qPCR). The maximum non-toxic concentrations of SGY against Vero and BV-2 cells were 382.804 μg·mL~(-1) and 251.78 μg·mL~(-1), respectively, and TC_(50) was 1 749.98 μg·mL~(-1) and 2 977.50 μg·mL~(-1), respectively. In Vero cell model, the half maximal inhibitory concentration(IC_(50)) of SGY against HSV-1 was 54.49 μg·mL~(-1), and the selection index(SI) was 32.12, with the mode of action of significantly inhibiting replication and directly inactivating HSV-1. RT-qPCR results showed that SGY markedly reduced the viral load in cells. The virus model group had significantly increased relative expression of TLR2, TLR3 and tumor necrosis factor receptor-associated factor 3(TRAF3) and reduced relative expression of TLR9 as compared with normal group, and after SGY intervention, the expression of TLR2, TLR3 and TRAF3 was decreased to different degrees and that of TLR9 was enhanced. The expression of inflammatory factors inducible nitric oxide synthase(iNOS), tumor necrosis factor-α(TNF-α), and interleukin-1β(IL-1β) was remarkably increased in virus model group as compared with that in normal group, and the levels of these inflammatory factors dropped after SGY intervention. In conclusion, SGY significantly inhibited and directly inactivated HSV-1 in vitro. In addition, it modulated the expression of TLR2, TLR3 and TLR9 related pathways, and suppressed the increase of inflammatory factor levels.
Animals ; Antiviral Agents/therapeutic use* ; Chlorocebus aethiops ; Herpes Simplex/pathology* ; Herpesvirus 1, Human/metabolism* ; Isoflavones ; Mice ; TNF Receptor-Associated Factor 3/pharmacology* ; Toll-Like Receptor 2/metabolism* ; Toll-Like Receptor 3/metabolism* ; Toll-Like Receptor 9/metabolism* ; Toll-Like Receptors/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Vero Cells ; Virus Replication

In this study, emotional stress-induced herpes simplex virus type 1(HSV-1) susceptibility model was employed to simu-late the pathological state of " depression-induced liver fire", and the protection effect of Qingre Xiaoyanning(QX) in clearing liver fire was investigated. BALB/c mice were randomly divided into a normal group, a HSV-1 group, a restraint stress + HSV-1 group,low-(0. 658 g·kg~(-1)) and high-dose(1. 316 g·kg~(-1)) QX groups, and an acyclovir group. Except for the normal group and the HSV-1 group, the mice in other groups received daily restraint stress for 6 h from day 3 of medication. On day 9 of medication, mice were anesthetized by isoflurane and infected intranasally with HSV-1. Survival rate, weight change, encephalitis symptoms, and eye injury of mice were recorded for 14 d after virus infection. Hematoxylin-eosin(HE) staining and immunohistochemical staining were used to detect pathological changes and HSV-1 antigen distribution. Plaque assay was performed to detect the titer of HSV-1. The protein ex-pression of ICP27 in the mouse brain was detected by Western blot. The experimental results showed that QX could increase the survival rate of HSV-1-infected mice loaded with emotional stress(P<0. 001), reduce the titer of HSV-1 in the mouse brain(P<0. 01), relieve brain inflammation(P<0. 05) and eye injury(P<0. 05), down-regulate the expression of ICP27 related to HSV-1(P<0. 05), and decrease the distribution of HSV-1 antigen in the mouse brain. The results demonstrated that QX significantly reduced the susceptibility to HSV-1 induced by emotional stress, which is expected to provide a theoretical basis for the treatment and preven-tion of HSV-1 infection and promote the clinical development and application of Chinese medicine effective in clearing liver fire.
Animals ; Capsules ; Herpes Simplex ; Herpesvirus 1, Human ; Mice ; Mice, Inbred BALB C ; Psychological Distress

PURPOSE: We report a case of herpes simplex keratitis after Descemet membrane endothelial keratoplasty (DMEK). CASE SUMMARY: A 67-year-old male underwent DMEK in his left eye due to pseudophakic bullous keratopathy. One week after DMEK, re-bubbling was performed due to partial detachment of Descemet's membrane at the corneal periphery. After re-bubbling, the cornea remained clear and the patient's visual acuity gradually improved. Two months after DMEK, the patient presented with mild discomfort and decreased visual acuity. The cornea showed an irregular, narrow dendrite with an epithelial defect and surrounding opacity. After confirming that Descemet's membrane was attached, the patient was started on oral valacyclovir for suspected herpes keratitis. Herpes simplex virus type 1 was eventually identified by polymerase chain reaction. The corneal lesion resolved after three weeks of antiviral treatment. CONCLUSIONS: Similar to penetrating keratoplasty, DMEK can trigger outbreaks of herpes simplex keratitis. Herpes simplex keratitis should remain on the clinician's differential diagnosis for patients who present with a corneal epithelial irregularity and decreased visual acuity following DMEK.
Aged ; Cornea ; Corneal Transplantation ; Dendrites ; Descemet Membrane ; Diagnosis, Differential ; Disease Outbreaks ; Herpes Simplex ; Herpesvirus 1, Human ; Humans ; Keratitis ; Keratitis, Herpetic ; Keratoplasty, Penetrating ; Male ; Polymerase Chain Reaction ; Visual Acuity

Chest pain in kidney transplant patients is usually caused by cardiac or pulmonary problems. However, it may be rarely caused by opportunistic esophageal infections. A 66-year-old female kidney transplant recipient was admitted because of chest pain. She had been treated with high-dose steroid and immunosuppressants for acute T-cell-mediated rejection. Cardiologic and pulmonary evaluations had normal results. Endoscopic examination revealed three clear ulcerative lesions in the esophagus. Histological and immunohistochemical staining of the endoscopic biopsy specimens revealed coinfection of herpes simplex virus and cytomegalovirus. The patient was treated with intravenous ganciclovir for 2 weeks. Her symptoms completely resolved, and follow-up endoscopy revealed complete healing of the previous ulcers. Viral esophagitis should be considered in the differential diagnosis in kidney transplant recipients presenting with chest pain.
Aged ; Biopsy ; Chest Pain ; Coinfection ; Cytomegalovirus ; Diagnosis, Differential ; Endoscopy ; Esophagitis ; Esophagus ; Female ; Follow-Up Studies ; Ganciclovir ; Herpes Simplex ; Humans ; Immunosuppressive Agents ; Kidney ; Kidney Transplantation ; Simplexvirus ; Thorax ; Transplant Recipients ; Ulcer

Herpes simplex virus (HSV) is a common pathogen, that causes a broad spectrum of diseases, ranging from minor skin infections to severe encephalitis and widespread infections. Acute retinal necrosis (ARN), one of the most serious manifestations of HSV infection, is defined as a rapidly progressing necrotizing retinopathy that presents discrete areas of circumferential retinal necrosis, along with signs of uveitis, vitreitis, and retinal vasculitis. We encountered a case of a female infant, born at 33 weeks of gestation with a body weight at birth of 2,080 g, who had ARN and encephalomalacia due to HSV infection. ARN associated with HSV infection should be suspected when nonspecific retinal exudates are observed in neonates, especially preterm infants.
Body Weight ; Encephalitis ; Encephalomalacia ; Exudates and Transudates ; Female ; Herpes Simplex ; Herpesvirus 2, Human ; Humans ; Infant ; Infant, Newborn ; Infant, Premature ; Necrosis ; Parturition ; Pregnancy ; Retinal Necrosis Syndrome, Acute ; Retinal Vasculitis ; Retinaldehyde ; Simplexvirus ; Skin ; Uveitis

A previously healthy 2.5-year-old male child presented with vomiting, diarrhea, and fever. During hospitalization he developed odynophagia and refusal to eat. His symptoms did not respond to acid suppressant therapy. He underwent upper endoscopy which showed severe inflammation, ulcerations and abundant necrosis. Histopathological features and serological testing were consistent with herpetic esophagitis. He had no history of recurrent infections or history of sick contacts. His immunological work up showed normal level of immunoglobulins and his White Blood Cells subpopulations were normal. His HSV serology was positive. The patient was started on acyclovir 5 mg/kg q 8 hours. He resolved his symptoms within 24 hours of treatment.
Acyclovir ; Child ; Diarrhea ; Endoscopy ; Esophagitis ; Fever ; Herpes Simplex ; Hospitalization ; Humans ; Immunoglobulins ; Inflammation ; Leukocytes ; Male ; Necrosis ; Serologic Tests ; Ulcer ; Vomiting

Disseminated neonatal herpes simplex virus (HSV) infection is a severe disease with a high mortality rate. Here, we report the patient presented with fulminant hepatic failure secondary to HSV infection followed by renal failure without any mucocutaneous symptoms. The patient recovered after treatment with acyclovir and peritoneal dialysis. This is the first known case of a patient in Korea who survived disseminated HSV infection with fulminant liver failure followed by renal failure without undergoing liver transplantation.
Acyclovir ; Hepatitis ; Herpes Simplex ; Humans ; Infant, Newborn ; Korea ; Liver Failure ; Liver Failure, Acute ; Liver Transplantation ; Mortality ; Peritoneal Dialysis ; Renal Insufficiency ; Simplexvirus

Streptococcus dysgalactiae has two main subspecies: S. dysgalactiae subsp. equisimilis (SDSE) and S. dysgalactiae subsp. dysgalactiae (SDSD). SDSE often colonizes and causes infections in humans; however, SDSD is an animal pathogen which often causes pyogenic infection in domestic animals. We present a case of meningitis with SDSD and herpes simplex virus in a 22-day-old newborn baby who had no exposure to animals.
Animals ; Animals, Domestic ; Bacteria ; Colon ; Herpes Simplex ; Humans ; Infant, Newborn ; Meningitis ; Simplexvirus ; Streptococcal Infections ; Streptococcus

Herpes simplex virus (HSV), including HSV-1 and HSV-2, is an important pathogen that can cause many diseases. Usually these diseases are recurrent and incurable. After lytic infection on the surface of peripheral mucosa, HSV can enter sensory neurons and establish latent infection during which viral replication ceases. Moreover, latent virus can re-enter the replication cycle by reactivation and return to peripheral tissues to start recurrent infection. This ability to escape host immune surveillance during latent infection and to spread during reactivation is a viral survival strategy and the fundamental reason why no drug can completely eradicate the virus at present. Although there are many studies on latency and reactivation of HSV, and much progress has been made, many specific mechanisms of the process remain obscure or even controversial due to the complexity of this process and the limitations of research models. This paper reviews the major results of research on HSV latency and reactivation, and discusses future research directions in this field.
Herpes Simplex ; virology ; Herpesvirus 1, Human ; physiology ; Humans ; Virus Activation ; physiology ; Virus Latency ; physiology ; Virus Replication