Abstract
Stroke has been regarded as an important health problem that produces a great impact on an annual healthcare expenditure, quality of life of the population, and the productivity of the country. However, the therapeutic efficacy is still limited because abundant of the stroke survivors live with the disability. Therefore, the efforts to reduce and to prevent the disability in the stroke survivors is essential and required. Our primary data in an animal model of stroke reveal that an anthocyanin-rich mulberry fruit can decrease brain infarcted volume, and brain dysfunction so we hypothesize that a consumption of an anthocyanin-rich mulberry fruit as an adjuvant therapy with the current therapy should decrease disability and dependency in the stroke survivors. The aim of this project is to proof this concept and to determine the consumption safety in the stroke volunteers. In the development of mulberry capsules for the application in volunteer testing, the product developed had a high humidity of 11.32%. It is slightly above the norm for most herbal products that should not exceed 10% except for some herbs because it is easy to have a microorganism accumulation in the high humidity condition. However, no contaminants were found, and the qualification of the developed mulberry product achieves the requirements according to the notification regarding to herbal pills of the Ministry of Public Health. In the developed mulberry capsule, anthocyanins especially cyaniding-3-glucoside (755.240±6.78 μg/g sample), were found in the highest amounts, followed by cyaniding-3-rutinoside (285.440±1.26 μg/g sample). In addition, chlorogenic acid (236.220±0.34 μg/g sample), gallic acid (108.500±0.24 μg/g sample), and quercetin (15.700 ±0.02 μg/g sample) also present, but in much smaller amounts. The developed mulberry capsule reveals the biological activities associated with stroke, especially the antioxidant, anti- inflammation, and neurotransmitters enhancement activities. When evaluating the storage time, the reduction of the phytochemical content, especially anthocyanin, and bioactivity when stored at 4 °C less were less than those at room temperature. Over a period of 3 months, the reduction of important phytochemicals and bioactivities remains in the range of about 20-25%. However, the longer they are stored, the lower their levels and bioactivities. Therefore, the process of extending product life must be developed in the case of commercial development. In the study regarding to the effect of mulberry capsule consumption in cerebrovascular disease subjects, the project is accredited by the Human Research Council, Khon Kaen University (HE631600), and it is registered in the Thai Clinical Trials Registry (TCTR) under the reference number TCTR20201222006. The research model is a three armed, randomized, double-blind, placebo-controlled experimental study. The volunteers are patients at the age between 18-85 years old with transient and ischemic stroke patients in the last 5-10 days that had weakness in one side of the body. All subjects are divided into 3 separated groups as following; the placebo group, and the groups that received mulberry fruit capsules at the doses of 1,000 mg, and 2,000 mg. Prior to the consumption of mulberry fruit capsule, and at 2 and 6 weeks of consumption, all volunteers are subjected to the assessments of the flowing parameters; body mass index, electrocardiography, blood pressure, respiration, clinical signs, and stroke severity with NIHSS score assessment. The assessments of hematological changes, and clinical chemistry are also performed to monitor the consumption safety of mulberry fruit capsule. The assessment of serum biomarkers consisting of MMP-9, S100β, vWF, and VCAM1 are performed prior to the consumption, and at a period of 1 and 6 weeks of consumption whereas the assessment of lactic acid producing bacteria which is regarded as probiotic bacteria particularly Lactobacillus spp and Bifidobacterium spp. In feces is performed prior to the consumption and at a 6 week-consumption period. The current results demonstrated that at 2 and 6 weeks of consumption, the Modified Rankin Scale (mRS) after stroke of volunteer in both placebo and mulberry fruit capsule show the significant reduction in mRS. However, the rates of change in mRS of the mulberry consumption groups both at 1,000 and 2,000 mg per day are significantly higher than that of placebo group. It has been revealed that the volunteers in a mulberry fruit consumption at a dose of 1,000 mg per day show a significant improvement of mRS only at 1 week after consumption whereas the volunteers in a mulberry fruit consumption at a dose of 2,000 mg per day show a significant improvement of the mentioned parameters after 1 week of consumption and this change is observed throughout the study period. In addition, the volunteers in the mulberry fruit treated group at a dose of 2,000 mg per day show the significant reduction of VCAM1 after 1 week of consumption, and the reduction of both VCAM1 and MMP9 are observed at 6 weeks of consumption. The current results suggest that mulberry fruit capsule at a dose of 2000 mg per day shows the good potential to reduce mRS in stroke and the underlying mechanism is partly associated with the reduction of VCAM1 and MMP9.
