| Objectives:Through the establishment of herpes simplex keratitis disease(HSK)model mice to the track lymphangiogenesis law in herpes simplex keratitis pathogenesis of mice’s cornea,to explore the relationship between herpes simplex virus keratitis and lymphangiogenesis.To provides a new strategy for the clinical prevention and treatment of herpes simplex virus keratitis.Methods:Corneal trephine was used to create regular circular defects in the central epithelium of BLA/C mice.The experimental group mice were inoculated with herpes simplex virus type 1(HSV-1)in the right eye,and the left eye was inoculated with an equal volume of DMEM medium.Another group of mice was inoculated with the same volume in both eyes The volume is the blank control group.Use slit lamp microscope to observe and evaluate the clinical symptoms,corneal opacity,corneal staining score of the experimental group and the blank control group at inoculated immediately,and 3D,5D,7D,10D,14D,28D,and 58D after virus infection.To evaluate the clinical symptoms of the eye in three aspects,take pictures and record the data;then take the mice cornea at each time point and fix it,and perform LVYE-1 and CD31 immunofluorescence staining to observe the position,morphology and distribution of corneal lymphatic vessels and blood vessels at each stage of the disease process,calculate the corneal blood vessel density,lymphatic vessel density,the distance from the end of the blood vessel or lymphatic vessel to the limbus in each infection stage.The data was statistically analyzed and compared to obtain the clinical symptoms at each infection stage during the onset of HSK,and summarize the characteristics and rules of lymphatic and blood vessel growth to obtain lymphatic vessel density and blood vessel density,The distance from the end of the blood vessel or lymphatic vessel to the limbus are related to clinical symptoms.Results:1).The right eye’s cornea can be repaired within 3 days after the formation of the corneal epithelial defect by the trephine,but the clinical symptoms of the eye gradually worsen afterwards,and the corneal ulcer and corneal edema form at 10 to 14 days.The severity of the clinical symptoms reaches a peak,and immune response mediated may occur.Corneal stromal inflammation,the condition progressed to 28 days after the virus inoculation,the ocular symptoms were relieved,the corneal epithelium was repaired,the corneal edema subsided,and the residual corneal opacity.Throughout the course of the disease,the scores of ocular clinical symptoms,corneal turbidity,and fluorescent staining all showed an upward trend from 3 days to 14 days after inoculation with HSV-1.The clinical symptoms and fluorescent staining scores of the eyes were 28 days after inoculation decreased,scarring of the cornea have residued,and corneal transparency reduced.The left eye of the mice and the blank control group had no clinical symptoms of infection during the entire course of the disease.2).Three days after infection with HSV-1,the lymphatic vessels tended to grow into the cornea earlier than the blood vessels.After the disease course progressed to 5D,the lymphatic vessels and blood vessels had grown into the cornea at the same time.The end of the blood vessel is closer to the corneal ulcer than the lymphatic vessel.After 28 days,the lymphatic and blood vessel are close to the corneal ulcer or have grown into the corneal ulcer.In the left eye of the mice and the blank control group no blood vessels and lymphatic vessels grew into the cornea during the entire observation period.3)The blood vessel density and lymphatic vessel density in the cornea of the right eye showed a gradual increase throughout the course of the disease.On the 7th day,the blood vessel density and the lymphatic vessel density were equivalent,and by the 28th day to the 58th day,both the blood vessel density and the lymphatic vessel density had no significant change in a stability.4)Within 5 days of infection,the distance from the end of the lymphatic vessel to the limbus is farther than the distance from the end of the blood vessel to the limbus.After that,the growth of blood vessels is larger than that of the lymphatic vessels and closer to the center of the cornea,and the distance from the end of the blood vessels to the limbus is greater than that from the end of the lymphatic vessels to the cornea.Until 28D days later,the end of the blood vessel tends to approach the center of the cornea and the ulcer tends to focus,and the lymphatic vessels can also approach the corneal ulcer or grow into it.5)The blood vessel density and lymphatic vessel density are similar to the growth pattern of the distance from the end of the blood vessel or lymphatic vessel to the limbus.They show a significant increase in the acute phase from 3 days to 14 days,and the upward trend slowly tends to increase from 28 days to 58 days,showing a similar trend with ocular clinical symptoms.In the acute phase,the infection is 14 days,blood vessel density,lymphatic vessel density,distance from the end of the blood vessel to the limbus,and the end of the lymphatic vessel to the limbus are consistent with the development and changes of the clinical symptoms of the eye,in this stage,the clinical symptoms of the eye were severe and the course of the disease progressed rapidly.The blood vessel density,lymphatic vessel density,the distance from the end of the blood vessel to the limbus,and the end of the lymphatic vessel to the limbus all increased significantly.When the disease progressed to the chronic stage and 28 days later,the clinical symptoms gradually relieved and stabilized,the blood vessel density,lymphatic vessel density,the distance from the end of the blood vessel to the limbus and the end of the lymphatic vessel to the limbus also basically formed no significant changes.Conclusions:1)The early corneal epithelium of herpes simplex virus keratitis can be repaired within 3 days,but the condition is still developing.2)The pathogenesis of herpes simplex keratitis is accompanied by the proliferation of blood vessels and lymphatic vessels in the cornea.3)During the onset of herpes simplex keratitis,the proliferation of lymphatics and blood vessels not be synchronized;4)In the course of herpes simplex keratitis,lymphatic vessels grow into the cornea earlier than blood vessels,and blood vessels grow faster than lymphatics and blood vessels in the later stages of the disease,the end is closer to the central corneal ulcer.5)In the course of herpes simplex virus keratitis,the acute stage of herpes simplex virus keratitis clinical symptoms develops rapidly,and the density of lymphatic vessels and blood vessels gradually increase,and the scale of the disease does not progress until the late stage of the disease.6)The growth of lymphatic vessels can aggravate the local immune inflammatory response and cause corneal damage,but the formation of corneal lymphatic vessels can also help reduce corneal edema.7)The regulation of blood vessels and lymphatic vessels in different periods is expected to become a target for the prevention and treatment of herpes simplex virus keratitis. |
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