Study On The Anti-anemia Activity And Safety Toxicological Evaluation Of Roselle Water Extract

Hibiscus Sabdariffa L.,also known as Luo Shen Hua and Hongtao K in China,is a herbal and edible plant.Its roots,seeds,leaves as well as calyxes can be used as medicine,but the most versatile part is its calyxes.Iron deficiency anemia?IDA?,the most universal type of anemias in the clinic,is a very common global disease which can cause a certain degree of harm to the human body.In fact,there are few studies on the role of Hibiscus Sabdariffa in mitigating IDA.Therefore,In this study the effect of aqueous extract of Hibiscus Sabdariffa?HS?on improving the symptoms of iron deficiency anemia in rats,preliminary analysis of extraction molecular mechanisms of antiserum activity and their related safety and toxicology studies were systematically investigated.The main contents and results are as follows:1. Evaluation of the effectiveness of HS against iron deficiency anemia in rats.The IDA rat model was based on low-iron feed and several exsanguinations.According to body weight and hemoglobin?HGB?levels,they were randomly divided into low iron group,Fe SO₄group,low,medium and high sample dose groups and normal group.The sample dose groups were given 250 mg/kg·bw,500 mg/kg·bw,1000 mg/kg·bw of HS,respectively.The positive control group was provided with 2mg/kg·bw ferrous sulfate solution,and the low iron group and normal control group were given normal saline of equal quantity.Intragastric administration was conducted once a day for 30 days.The relevant indicators of iron deficiency anemia were recorded to evaluate the effect of HS on IDA.The results showed that the HGB levels of the dose groups were significantly higher than that of the low iron group,which were significantly different from the normal group,but both were within the normal range;while the content of free erythrocyte protoporphyrin?FEP?and serum transferrin receptor?s Tf R?were markedly lower than that of the low iron group.To sum up,the different extract dosage administered by the rats can improve the iron deficiency anemia of rats to some extent.2. Preliminary analysis of the molecular mechanism of the extract anti-anemic activity.IDA rats were treated with different doses of HS.Then,Q-PCR and Western blot were used to detect the expression of divalent metal transporter 1?DMT-1?and ferroportin?FPN?,hephaestin?Hp?and their m RNA in the duodenum and the expression levels of hepcidin and its m RNA in liver of the rats.The effects of the extract exerting upon related genes and proteins were tentatively analyzed here.The results showed that HS inhibited the relative expression of DMT-1,FPN-1,Hp and their m RNA in the duodenum of IDA rats.The expression of Hp as well as its m RNA was only affected by the extracts within a certain dose range.Low extract concentration promoted the expression of Hepcidin-25 in the liver of IDA rats,however,high concentration inhibited the expression of Hepcidin-25.All dose groups showed inhibiting effect on the related m RNA expression.3. Safety and toxicology evaluation of HS.The safety of HS was assessed based on the acute toxicity test and 30-day test.The results of the acute toxicity test showed that LD₅₀of HS was greater than 5000 mg/kg,which was considered to be virtually non-toxic.The results of the 30-day test indicated that the indicators of the rats in each dose group?general health status,body weight,feed intake,hematology and serum biochemical main indicators,major organ weight and coefficients?,changed within the normal range,showing no abnormality when compared with the normal control group.No pathological changes were observed in the structure and morphology of the major organs of each test group.It indicated that the HS can be consume safely.In summary,the aqueous extract of HS has a good effect on iron deficiency anemia of rats,and can regulate the expression of genes and proteins related to iron absorption and transport to a certain extent.It is safe and non-toxic.This provides a theoretical basis for the safety of Hibiscus Sabdariffa-related health care products.