Methods for attenuating parasite virulence
A technology of parasites and malaria parasites, applied in the direction of pharmaceutical formulations, organic active ingredients, and resistance to vector-borne diseases, etc., can solve problems affecting the infection process and disease onset process
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[0038] Example 1 Host feeding affects survival and parasite load. C57BL / 6 mice (5-8 weeks old, body weight 20-28 g) were allowed free access to water and food, or placed in a calorie restricted (CR) group. Mice in the CR group were given 60-70% of the food per day relative to the food consumed by the ad libitum control group (AL). Food intake in both groups was measured once a day, and body weight was measured at least 3 times a week. When 15–20% body weight loss is achieved, adjust the daily food amount allocated to CR mice or rats to stabilize lower body weight for the remainder of the experimental period. By intradermal (i.d.) injection of 5 x 10 3 Freshly dissected Plasmodium berghei sporozoites ( figure 2 ) or by intraperitoneal (i.p.) injection of 10 6 Plasmodium berghei-infected erythrocytes were used to infect mice, and Plasmodium berghei-infected erythrocytes were obtained from the previous passage of C57BL / 6 mice ( image 3 ). House 3-5 mice per cage.
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example 2
[0040] Example 2 AMPK activating compounds mimic CR-mediated AMPK activation. Western blot analysis of AMPK phosphorylation in liver homogenates from uninfected C57BL / 6 mice in the AL and DR fed groups and uninfected mice one hour after salicylate or vehicle injection, see Figure 4 . The liver was dissolved in ice-cold lysis buffer (50 mM HEPES, 150 mM NaCl, 10 mM NaF, 1 mM sodium pyrophosphate, 0.5 mM EDTA, 1 mM DTT, 1% octylphenol polyethylene glycol ether (triton), 1 mM Na 3 VO 4 , 250mM sucrose, protease inhibitor cocktail and phosphatase inhibitor). The total protein content in each homogenate was measured by the Bradford Assay (Bio-Rad) according to the manufacturer's instructions. Dissolve 50 μg of total liver lysate on 8% SDS-PAGE or Any kD TM Precast gels (Bio-Rad) and transferred to nitrocellulose membranes using a standard wet transfer with 1× Tris-glycine buffer (containing 20% methanol) at a constant voltage of 100 V for 2 hours, or using Gel Transfer S...
example 3
[0042] Example 3 Reduction of Plasmodium liver stage infection by AMPK activating compounds. 72 hours post infection h.p.i. of C57BL / 6 mice infected with freshly dissected P. Relative parasitaemia in C57BL / 6 mice was analyzed by flow cytometry (as described in Example 1). One hour before infection, salicylate (300 mg / Kg, Sigma No. 71945) was administered once daily by intraperitoneal injection. Metformin (500 mg / Kg, Sigma No. D150959) was provided in drinking water one week before and during infection. see results Figure 5 .
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