Effect Of Charred Radix Et Rhizoma Rhei In A Laser-Induced Choroidal Neovascularization Murine Model

Purpose:A pharmaceutical composition(patentno. WO2012079419) exhibited favorable outcomes in a clinicaltrial of wet age-related macular degeneration. The aims of thepresent study were to explore the effects of one compositioncomponent, charred Radix et Rhizoma Rhei(CRRR), in alaser-induced choroidal neovascularization(CNV) murine model.Methods:1、A total of 30 eight-week-old C57BL/6 mice weresubjected to diode laser treatment,and CNV was inducedby rupturing the Bruch’s membrane.2、The mice were thenrandomly divided into two groups: the CRRR-treated groupthat was administered CRRR water extract(concentration,0.6 g/100 ml; dose, 1 ml/0.1 kg twice a day for 21 days); and thecontrol group that was treated with saline(dose, 1ml/0.1 kgtwice a day for 21 days).3、The retinal tissue was subjected toquantitative polymerase chain reaction(qPCR)and western blotanalysis to determine the expression levels of interleukin-10(IL-10)and vascular epithelial growth factor(VEGF) at dayseven following laser treatment.4、At weeks 2 and 3 after lasertreatment, fundus fluorescein angiography(FFA) was performedand graded to assess the severity of lesion leakage.5、RPE-choroid-sclera flat mounts were prepared for three-dimensional confocal-microscopy at day 22 after laser treatment.Results:1、At days 14 and 21 after laser treatment, no statistically significant differenceswere observed between the clinically relevant lesions of the CRRR-treated and control mice.2 、 At days 14 and 21 after laser treatment,CNV volumes were not found to be significantly different between the CRRR?treated and control mice.3 、 At days 7 following laser treatment,the expression levels of IL?10 were significantly increased in the CRRR-treated mice(P<0.05).4、At days 7 following laser treatment,however, no statistically significant differences were observed between the VEGF expression levels of the CRRR-treated and control mice.Conclusion:1、 CRRR appeared to some significantly inhibit CNV in this murine model.2、 The function of CRRR inthe pharmaceutical composition may be due to the effects of IL-10 and a synergistic effect with other components of the composition.