Application of RSK signal channel inhibitors to restraining chlamydia trachomatis infection
A technology of Chlamydia trachomatis and signaling pathway, which is applied in the field of biomedicine and can solve problems such as no reports of Chlamydia infection-assisted targeted host therapy.
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Embodiment 1
[0059] Example 1 LJH685 and LJI308 inhibit Chlamydia infection
[0060] The purpose of this example is to clarify the role of RSK signaling pathway inhibitors LJH685 and LJI308 in Chlamydia infection. Hela cells were cultured and inoculated with Chlamydia trachomatis type D. Set up positive and negative controls at the same time. Centrifuge the inoculated 24-well culture plate at 35°C and 1500g for 1h. After centrifugation, suck all the inoculated sample solution, replace each well with DMEM medium containing 1 μg / ml cycloheximide, add 80 μM LJH685 and 20 μM LJI308, 5% CO 2 1. Cultivate for 48 hours under the condition of constant temperature and humidity at 35°C, and then observe the results. Chlamydia inclusion bodies were observed under an inverted microscope by iodine staining and immunofluorescence. Randomly count 20 400-fold visual fields, and calculate the infection rate of chlamydia. 48 hours after infection, collect 80 μM LJH685 and 20 μM LJI308-treated cells wit...
Embodiment 2
[0075] Example 2 LJH685 and LJI308 have similar effects in different cell lines
[0076] The purpose of this example is to clarify the role of LJH685 and LJI308 in different cells. Rat fibroblast McCoy cells and green monkey kidney Vero cells were cultured and inoculated with Chlamydia trachomatis type D. Set up positive and negative controls at the same time. Centrifuge the inoculated 24-well culture plate at 35°C and 1500g for 1h. After centrifugation, suck all the inoculated sample solution, replace each well with DMEM medium containing 1 μg / ml cycloheximide, add 40 μM LJH685 and 20 μM LJI308, 5% CO 21. Cultivate for 48 hours under the condition of constant temperature and humidity at 35°C, and then observe the results. Chlamydia inclusion bodies were observed under an inverted microscope by iodine staining and immunofluorescence. Randomly count 20 400-fold visual fields, and calculate the infection rate of chlamydia. 48 hours after infection, collect 80 μM LJH685 and ...
Embodiment 3
[0092] Example 3 LJH685 and LJI308 have similar effects on different serotypes of Chlamydia infection
[0093] The purpose of this example is to clarify the role of LJH685 and LJI308 in different serotypes of Chlamydia infection. The specific experimental methods and results are as follows:
[0094] 1. Experimental method:
[0095] Hela cells were cultured and inoculated with Chlamydia trachomatis. Iodine staining method and immunofluorescence method were used to observe Chlamydia inclusion bodies under an inverted microscope, calculate the infection rate of Chlamydia, reinoculate Chlamydia trachomatis, and count the number of infectious Chlamydia.
[0096] 2. Experimental results:
[0097] The result is as Figure 15-16 :
[0098] Figure 15 : The role of LJH685 and LJI308 in L1 Chlamydia infection: Hela cells were infected with L1 serotype Chlamydia trachomatis, and found that LJH685 and LJI308 can inhibit the infection of L1 serotype Chlamydia. Immunofluorescence obs...
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