动物造模,药效评价,生殖器疱疹 z-bio 2024-07-24 493

　　Genital herpes is a sexually transmitted disease caused by the human herpes simplex virus, with 90% of the pathogens being HSV-2. Injecting HSV-2 virus solution into the vagina of animals is a common method for establishing animal models of genital herpes. The genital herpes model of female guinea pigs is considered the best model for studying latent infection of herpes virus and new antiviral drugs and vaccines. Meanwhile, through the establishment of the model, the pathogenic mechanism of herpes virus infection can also be studied, and the immune mechanism of the body against viruses can be investigated.

　　1、 Establishment and Research Methods of Genital Herpes Animal Model

　　At present. The animal models of genital herpes used for pharmacological research reported abroad mainly include mouse, monkey, and guinea pig models. Although mice can develop HSV-2 genital infections, they rarely show visible lesions and are prone to death; The vaginal HSV-2 infection pattern of monkeys is similar to that of humans, making it an ideal primate model, but its use is limited by its high cost; Guinea pig vaginal HSV-2 infection can not only present clinical manifestations similar to human genital herpes initial and recurrent infections, but also establish latent infections in the nervous system with low mortality and low cost. Therefore, establishing a genital herpes animal model using female guinea pigs is currently a feasible and ideal choice.

　　1. Method for establishing a model of primary genital herpes in guinea pigs: First, clean the guinea pig's external genitalia with physiological saline, rub the vagina several times with a dry cotton swab, and then gently tap the guinea pig's external genitalia with a plum blossom needle to make it congested and slightly oozing blood. Clean the external genitalia with a clean cotton swab. Next, use a 1ml syringe to aspirate 0.15ml of human herpes simplex virus-2Sav strain virus solution, and insert a gavage needle into the guinea pig's vagina 3-4 cm away. Inject the virus solution into the vaginal vault and slowly withdraw. Finally, insert a gelatin sponge into the vaginal opening to maintain the virus solution. A small amount of venom drips onto the external genitalia from the needle, and is gently spread evenly with a glass rod to allow the virus to penetrate the skin. Observe the appearance of skin lesions.

　　2. Establishment method of guinea pig HSV-2 recurrent infection model: About 20 days after the initial infection in guinea pigs recovered, cyclophosphamide and physiological saline were prepared into a solution at a concentration of 50mg/ml and injected intraperitoneally into guinea pigs at a dose of 1ml/kg. Considering the long modeling period of cyclophosphamide, after 10 days of injection of cyclophosphamide, ultraviolet radiation (wavelength 270-320nm) was used, and the dose was adjusted to 7000 μ W/cm2. After anesthesia, guinea pigs were exposed to ultraviolet radiation for 10 minutes for a total of 5 days to induce the appearance of skin lesions in the perineum. Observe changes in the external genitalia. Generally, blisters appear after 5 days.

　　3. The current literature mostly refers to Kern et al.'s method for symptom scoring: by observing the degree of lesions in the external genitalia of each guinea pig, a score is given: 0 is asymptomatic; 0.5 is only red and swollen without blisters; 1.5 is a single small blister (≤ 2mm); 2.0 is a single large blister (>2mm); 2.5 refers to multiple small blisters and/or vaginal ulcers (bleeding); 3.0 represents multiple large blisters; 3.5 indicates severe swelling of the external genitalia; 4.0 represents the fusion of multiple small (large) blisters; 4.5 is paralysis of the hind limbs; 5.0 is an external genital ulcer.

　　2、 Application research on animal model of genital herpes

　　One important use of establishing a genital herpes animal model in the research of new anti herpesvirus drugs is for pharmacological studies. Rose et al. found through a guinea pig model that oral administration of the newly synthesized dipeptide SCV-07 (γ - D-glutaryl-L-l-transerine) has potential antiviral and immunomodulatory activity, and has a good therapeutic effect on recurrent HSV-2 infection. Moreover, taking it on an empty stomach in the morning has a better effect, providing a good choice for the treatment of recurrent genital herpes. Kratz et al. have shown that both gallic acid and pentanylgallic acid have local inhibitory effects on herpes virus replication. Domestic scholars have also conducted research on the mechanism of traditional Chinese medicine treatment for recurrent genital herpes through guinea pig animal models. The results show that traditional Chinese medicine antiviral capsules have a preventive effect on HSV.2 infection and damage to ganglia in recurrent genital herpes animal models.

　　Another important application of establishing a genital herpes animal model in the development of herpes virus vaccines is the development of herpes vaccines. The development and efficacy evaluation of HSV vaccines through animal models is currently a research hotspot. At present, some of these vaccines have entered the clinical evaluation stage, such as subunit vaccines, attenuated live vaccines, DNA vaccines, etc.

　　The DNA vaccine is currently the most extensively researched and promising HSV vaccine for development and use. Among them, the most researched vaccines are HSVgD vaccine and HSV gB vaccine. Bemstein et al. found that using cationic liposomes The HSV-2gD2 recombinant protein subunit vaccine with DNA complex as adjuvant, compared with the single gD2 vaccine and the HSV-2gD alum MPL vaccine, can effectively prevent HSV infection, significantly reduce the replication of the virus in the vagina, reduce the latent infection of the virus, and significantly reduce the clinical incidence rate. Reszka et al. demonstrated through guinea pig model studies that a new recombinant HSV-2dl5-29-41.1 has excellent anti herpesvirus activity and has the potential to become a vaccine for the development of HSV-2. Hoshino et al.'s study showed that dl5-29-41L and dl5-29 have equivalent immunity to HSV-2 infection in guinea pigs, while dl5-29 is safer for animals with immune dysfunction. Although these vaccines have achieved good results in animal experiments, they cannot effectively prevent the recurrence and infection of the virus in humans. Therefore, in the future, by establishing more comprehensive animal models, it is expected to develop more effective vaccines for humans.

     3. Application in the study of the pathogenic mechanism of herpes virus: Establishing a genital herpes animal model can provide a more in-depth understanding of the pathogenic mechanism of herpes virus. Crostarosa et al. established a unique animal model of rhesus monkeys, and the results showed that vaginal HSV-2 infection in rhesus monkeys has a synergistic effect on HIV infection. Zariffard et al.'s study also found that vaginal HSV-2 infection in mice promotes HIV infection in mice. This study suggests that this animal model can also be used to investigate the mechanism of human HSV-2 infection in promoting HIV susceptibility and explore possible preventive measures. Thapa et al. demonstrated through a mouse model that CXCL9 or CXCL10 play a crucial role in mobilizing effector cells such as NK cells and CD8+T cells to the site of infection in the host after HSV-2 infection. Gill et al. conducted experiments on a mouse HSV-2 infection model, and the results showed that IL-15 has inherent antiviral and immune effects. Once IL-15 is lacking, the body's adaptive immune response cannot provide defense against HSV-2 infection. Wilson et al. found that damage or incompleteness of vaginal epithelial cells in mice increases the probability of HSV-2 infection and also increases the risk of HIV infection. Pennock et al.'s study showed that estradiol can enhance the efficacy of mouse genital herpes vaccines, providing an idea for the development of other vaccines, but also suggesting whether there are differences in vaccine efficacy between genders. Interestingly, Pennock et al. also found that the drug methylenedioxymethamphetamine (ecstasy) can increase the susceptibility of mice to genital herpes, and there have been no similar studies reported in humans.

　　In summary, establishing an animal model of HSV-2 infection is of great significance for further studying the pathogenesis of genital herpes, as well as for the development of herpes virus vaccines and new drugs. Although there are certain limitations in research, such as the effectiveness and impact of vaccines on different genders, and whether new drugs can exhibit the same effects in humans as in animal models, further research and exploration are needed. It is believed that with the continuous advancement of experimental technology in the future, the establishment of animal models can play a greater role and better serve humanity.