Therapeutic Effect Of Compound Tufuling Granules In Hyperuricemia At Remission Of Gout And The Mechanism Of Reducing Uric Acid Based On ABCG2

Objective1.To evaluate 12-week clinical efficacy and safety of Compound Tufuling Granules in patients of hyperuricemia at remission of gout with syndrome of blockage of phlegm and blood stasis, and to provide drug selection and basis for clinical treatment.2. Based on uric acid transport protein ABCG2 to explore the urate lowering, mechanism of Compound Tufuling Granules, and to provide the experimental basis for clinical application.Method1. Clinical research:66 subjects in hyperuricemia at remission of gout with syndrome of blockage of phlegm and blood stasis were included, and were randomized to Compound Tufuling Granules group or Benzbromarone group by 1:1 ratio, which were given corresponding drug therapy for 12-week. Evaluate the efficacy of traditional Chinese medicine and Western medicine, as well as the safety evaluation.2. Experimental study:48 SPF male SD rats were randomly divided into six groups, as the control group, model group, Benzbromarone group, high-dose Compound Tufuling Granules group, Medium-dose Compound Tufuling Granules group, and Low-dose Compound Tufuling Granules group. Except the control group, the rest of the groups were given potassium oxonate and hypoxanthine to induce hyperuricemia. Modeling agent administered once daily and continued eight days. Four days after the start of the experiment, at 30 minites later of given modeling agent, Benzbromarone group, High-dose, Medium-dose and Low-dose Compound Tufuling Granules group were given corresponding doses of drugs orally. Model group and the control group were only given the normal saline. The treatments were given once a day, a total of five days of treatment. Eight days after the start of the experiment, at 3 hours later of drug treatments, serum uric acid(sUA), serum creatinine and blood urea nitrogen were tested. Except the Benzbromarone group, ABCG2 mRNA expression of the rest of groups were detected with qRT-PCR method and ABCG2 expression of the rest of groups were detected with Western Blot and immunohistochemical methods in liver, kidney and small intestine.Result1.63 patients completed the study, including 32 cases of Compound Tufuling Granules group and 31 cases of Benzbromarone group.2. At the end of the 12 week treatment, the percentages of patients achieved total effective standard of Tradition Chinese Medicine in Compound Tufuling Granules group and Benzbromarone group were 90.625% and 93.548% respectively, and there was no significant difference (P=1.000).3. At the end of the 12 week treatment, the percentages of patients with sUA<420umol/L in Compound Tufuling Granules group and Benzbromarone group were 84.4% and 90.3%. There was no significant difference between the two groups (P=0.708).4.The percentage of patients with sUA<420umol/L in Compound Tufuling Granules group was 50.0% at week 4 and 75.0% at week 8. That percentage of Benzbromarone group was 67.7% at week 4 and 74.2% at week 8,There was no significant difference between the two groups at week 4 or 8(P=0.203,1.000).5.SUA levels of the Compound Tufuling Granules group in week 4,8 and 12 were significantly lower than those of the baseline sUA (P=0.000、0.000、 0.000). SUA levels of the Benzbromarone group in week 4,8 and 12 were significantly lower than those of the baseline sUA(P=0.000、0.000、0.000). There was no significant difference between the two groups at week 4 or 8 or 12 (P=1.000、1.000、1.000)6. The percentages of patients with sUA<300umol/L in Compound Tufuling Granules group and Benzbromarone group were 34.4% and 45.2% at week 4, and there was no significant difference (P=0.446). The percentages of patients with sUA<300umol/L in Compound Tufuling Granules group and Benzbromarone group were 50.0% and 51.6% at week 8, and there was no significant difference (P=1.000). That percentages of two groups were 62.5% and 67.7% at week 12 (P=0.793).7.During the 12 week period,0 patient (0%) with acute gout arthritis occurred in Compound Tufuling Granules group,and 2 patients (6.5%) patients with acute gout arthritis in Benzbromarone group. There was no significant difference between the two groups (P=0.238).8. During the 12 week period, there was no clinical significance of the abnormal test and check. No adverse events occurred.9. The level of sUA in the model group was significantly higher than that in the control group (P=0.000). The level of sUA in the High-dose Compound Tufuling Granules group or Benzbromarone group was equal to the control group (P=1.000、0.905). The level of sUA in the Benzbromarone group or High-dose Compound Tufuling Granules group or Medium-dose Compound Tufuling Granules group or Low-dose Compound Tufuling Granules group was lower than the model group (P=0.000、0.000、0.001、0.003). The level of sUA in the High-dose Compound Tufuling Granules group was lower than the Medium-dose Compound Tufuling Granules group (P=0.002), and the level of sUA in the Medium-dose Compound Tufuling Granules group was lower than the Low-dose Compound Tufuling Granules group (P=0.014)10. There were no significant differences in serum creatinine and urea nitrogen levels in each group (P=0.563、0.697)11. QRT-PCR method was used to detect the expression of ABCG2 mRNA in liver, kidney and small intestine of rats in each group. The expression of liver ABCG2 mRNA in model group was significantly lower than that in the control group (P=0.000). The expression of liver ABCG2 mRNA in high, medium and low dose Chinese medicine group was significantly higher than that in model group (P=0.000、0.000、0.000). The expression of liver ABCG2 mRNA in high dose Chinese medicine group was higher than that in medium dose Chinese medicine group (P=0.000). The expression of liver ABCG2 mRNA in medium dose Chinese medicine group was higher than that in low dose Chinese medicine group(P=0.000). The results of ABCG2 mRNA expression in the kidney and small intestine of rats in each group were similar to the liver.12. Western Blot method was used to detect the expression of ABCG2 in liver, kidney and small intestine of rats in each group. The expression level of liver ABCG2 in the model group was significantly lower than that in the control group (P=0.000). The expression levels of liver ABCG2 in the high, medium and low dose Chinese medicine group were higher than those in the model group(P=0.000、 0.000、0.000).The expression levels of liver ABCG2 in the high, medium and low dose Chinese medicine group were in order to reduce (F＜0.05). The results of ABCG2 expression in the kidney or small intestine of rats in each group were similar to the liver.13. The expression of ABCG2 in liver, kidney and small intestine of rats was detected by immunohistochemical methods. The expression of ABCG2 in the liver of the control group was higher than that in the model group, and the expression of ABCG2 in the liver of each dose Chinese medicine group was higher than that in the model group. The expression of kidney ABCG2 in the control group was higher than that in the model group, and the expression of kidney ABCG2 in each dose Chinese medicine group was higher than that in the model group. The expression of ABCG2 in the small intestine of the control group was higher than that in the model group, and the expression of ABCG2 in the small intestine of each dose Chinese medicine group was higher than that in the model group.Conclusion1. Compound Tufuling Granules is an effective and safe approach in the treatment of hyperuricemia at remission of gout with syndrome of blockage of phlegm and blood stasis, which can provide drug selection and basis for clinical treatment.2. Compound Tufuling Granules have good urate lowering effect, and the urate lowering effect would increase gradually by the increasing doses of Compound Tufuling Granules. The urate lowering mechanism of Compound Tufuling Granules might be promoting the expression of transport protein ABCG2 to increase the excretion of uric acid.