The Therapeutic Effect Of Compound Tufuling Granules On Hyperuricemia And Its Effect On MiR-34a And URAT1Expression In HK-2Cells

Objective1. By observing the changes of uric acid concentration clinical symptoms and safety indexes, To evaluate the clinical effect and security of Compound Tufuling Granules on atients with hyperuricemia.2. HK-2cells were cultured by a routine method and transfected with miR-34a mimics. To explore the influence of the medicine-containing serum of Compound Tufuling Granules on the expression of miR-34a and URAT1mRNA in HK-2cells, that to search for the mechanism of reducing uric acid.Methods1. Clinical observation:105patients with hyperuricemia were randomly divided into control group, the Benzbromarone group and the Compound Tufuling Granules group, the observation period is30days. During the period of treatment, the patients in the control group only to a unified health education, the other two groups were given oral Benzbromarone capsules and Compound Tufuling Granules. The clinical symptoms, physical signs and laboratory data before and after treatment were noted and compared.2. Cell experiment:Cultured HK-2cells were divided into two groups: miR-34a transfection group and Control group. miR-34a transfection group were transfected with miR-34a mimics. Control group were transfected with mimic Negative Control solution. miR-34a and URAT1mRNA expression levels of HK-2cells was measured by qRT-PCR method. High expression of miR-34a HK-2cells were divided into five groups:High-dose containing serum of Comound Tufuling Granules group, Medium-dose containing serum of Compound Tufuling Granules group, and Low-dose containing serum of Compound Tufuling Granules group, SD male mice serum group, and Control group. To detect the expressions of miR-34a and URATl mRNA in HK-2cells with qRT-PCR.Results1. The significant decrease of the degree of uric acid was found in the groups of Hyperuricemic patients after treating with Benzbromarone and Compound Tufuling Granules (P<0.05). The total effective rate was85.71%in Compound Tufuling Granules group,91.18%in Benzbromarone group, and36.36%in Control group. The effect in Compound Tufuling Granules group and Benzbromarone group was better than that in Control group (P<0.0167). There was no statistically significant difference between Compound Tufuling Granules group and Benzbromarone group (P>0.0167).2. The total effective rate in the clinical symptoms of patients was94.12%in Compound Tufuling Granules group,68.57%in Benzbromarone group, and36.36%in Control group. The effect in Compound Tufuling Granules group was better than that in other group.(P<0.0167). The clinical symptoms of patients in Compound Tufuling Granules group, included lassitude, heaviness of limbs, phlegm, loss of appetite, loose stool were improved significantly (P<0.05).3. The blood cholesterol, triglycerides and LDL cholesterol was improved better in Compound Tufuling Granules group as in Benzbromarone group and Control group (P<0.05).4. During treatment, none of the Patients in each group has adverse reactions. The laboratory data of patients in Compound Tufuling Granules, included blood routine, liver function, renal function and urine routine, were no significant difference after treatment (P>0.05).5. Detected by qRT-PCR, miR-34a transfection group was significantly increased by the amount of miR-34a expression compared with the control group, the difference was statistically significant (P<0.05). miR-34a transfection group was significantly decreased by the amount of URAT1mRNA expression compared with the control group, the difference was statistically significant (P<0.05).6. Detected by qRT-PCR, the expressions of miR-34a in High expression of miR-34a HK-2cells were increased significantly in each dose Compound Tufuling Granules group than SD male mice serum group, and Control group (P<0.05), the mRNA exPressions of URAT1were decreased significantly in each dose Compound Tufuling Granules group than SD male mice serum group, and Control group(P <0.05). Gonelusion1. Compound Tufuling Granules can reduce serum uric acid, cholesterol, triglycerides and LDL cholesterol levels, improve a part of clinical symptoms effectively in Hyperuricemic patients, while it is safety.2. Compound Tufuling Granules can effectively increase the expressions of miR-34a, decrease the mRNA expressions of URAT1in HK-2cells. Its mechanism of reducing uric acid might be increasing the expressions of microRNA to inhibit the expressions of URAT1.