Expression Of MiRNAs In Plasma Of Patients With Chronic Gout And The Intervention Effect Of Tiger - Tong - Tong Mixture On The Expression Of Differentially Expressed MiRNAs

Purpose: Micro RNA gene chip technique was applied to detect the expression spectrum of micro RNAs(mi RNAs) in the plasma of patients with chronic gout arthritis, and to screen the differential expression of mi RNAs in the plasma of patients and healthy people. Fluorogenic quantitative PCR(RT-q PCR) technique was applied to check the differential expression of mi RNAs. Chuanhutongbi mixture in the treatment of chronic gouty arthritis clinical efficacy and the intervention effect of mi RNAs differentially expression will be studied simultaneously. Explore the medical mechanism of mi RNAs in chronic gout therapy. Study on targets of Chuanhutongbi mixture in the treatment of chronic gout. Provide new clinical and molecular basis for Chinese traditional medicine for the prevention and treatment of chronic gout diseases.Method:1. Selecting 4 samples of fasting blood plasma in chronic gout arthritis patients and 4 samples of contrast healthy people, use Trizol method to extract RNA. Use LNATM microarray mi RNAs exqion high throughput detection to screen mi RNAs expression spectrum. Use cluster analysis software to analyze the differential expression of mi RNAs. 2. Expanding the sample of patients with chronic gout arthritis up to 60 cases, and the sample of healthy group up to 30 cases, use RT-q PCR method to check and analyze the partial differential expression of mi RNAs. 3. Adopt randomized, double-blind, double simulation, parallel contrast studying method, according to figure random distribution method the 195 cases of chronic gouty arthritis patients will be divided into Chuanhutongbi mixture group, allopurinol group and contrast group. Collecting patients plasma based on before treatment and 4 weeks after treatment and 8 weeks after treatment, use RT-q PCR to detect partial difference of mi RNAs. Adopt Enzyme linked immunosorbent assay(ELISA) method to determine the level of chemotaxis factor 2(CCL2) and chemotaxis factor 8(CXCL8). Analyze Chuanhutongbi mixture difference expression of mi RNAs and intervention effect of chemokine CCL2, CXCL8.Result1. In chronic gouty arthritis patients plasma samples, 48 mi RNAs were differentially expressed. The obviously up regulating expression and the value equal or more than 1.5 times compare with contrast group are 36 mi RNAs, which were mi R-4748, mi R-4726-5p, mi R-10a-3p, mi R-302a-3p, mi R-4251, mi R-5580-5p, mi R-639, mi R-4764-5p, mi R-631, mi R-4531, mi R-1284, mi R-H8-5p, mi R-198; mi R-4466, mi R-874-3p, mi R-5190,mi R-205-3p, mi R-204-5p, mi R-183-3p, mi R-H1, mi R-1471, mi R-660-3p, mi R-92a-2-5p, mi R-1255 a, mi R-505-5p, mi R-765, mi R-4780, mi R-4324, mi R-4738-3p, mi R-4800-5p, mi R-B1-5p, mi R-4455, mi R-5004-3p, mi R-4632-3p, mi R-4634. The obviously down regulating expression and the value less than 1.5 times compare with contrast group are 12 mi RNAs, which were mi R-548 l, mi R-361-5p, mi R-326 mi R-151a-3p, mi R-486-5p, mi R-769-5p, mi R-130b-3p, mi R-423-3p, mi R-4288, mi R-4732-5p, mi R-584-5p, mi R-339-5p. 2. Furthermore, RT-q PCR analysis showed that the expression levels of mi R-486-5p, mi R-361-5p and mi R-339-5p in plasma of patients with chronic gout arthritis were significantly lower than those in healthy contrast group, and the diversity comply with statistically significance(p<0.05), and the results were consistent with the results of mi RNAs gene chip. 3, Clinical trial results show that no significant difference among Chuanhutongbi mixture group, allopurinol group and contrast group of patients baseline characteristics(P >0.05). Comparing the diffrent value among the Three groups after 4 weeks of treatment, Chuanhutongbi mixture and allopurinol groups are much better than contrast group in up-regulated in plasma mir-486-5p and mi R-361-5p and reduce plasma levels of CCL2 aspects. Regarding to up-regulating plasma mir-339-5p and reduce plasma levels of CCL8 aspect, Chuanhutongbi mixture is better than allopurinol and contrast group.Difference comparison after 8 weeks of treatment among three groups, Chuanhutongbi mixture and allopurinol group could do better job on up-regulating plasma mir-486-5p and mi R-361-5p level compared with the contrast group. Chuanhutongbi mixture were better in up-regulating plasma mir-339-5p and reducing the level of blood uric acid compare with allopurinol group and the contrast group. Intentionality analysis showed that Chuanhutongbi mixture and allopurinol group can significantly reduce chronic gouty arthritis of acute exacerbation rate comparing with the contrast group. And what’s more, Chuanhutongbi mixture has less side effects comparing with allopurinol group. 4. Chronic gouty arthritis patients plasma mi R- 486-5p and mi R- 361-5p level instead of pain scores and c-reactive protein(CRP) and blood sedimentation were significantly negative correlation.Conclusion: 1. In plasma samples of chronic gout arthritis The expression of mi R-502-5p, mi R-4748 and other 36 mi RNAs shows up regulating, mi R-339-5p、mi R-486-5p、mi R-361-5p and other12 mi RNAs shows down regulating. 48 differential mi RNAs may play an important role in the pathogenesis of chronic gout arthritis. 2. Plasma levels of mi R-486-5p and mi R-361-5p were correlated with the severity of inflammation in patients with chronic gout arthritis, which could be used as a biological indicator to judge the severity and prognosis of the disease. 3, Chuanhutongbi mixture and allopurinol almost have the same clinical efficacy in the treatment of chronic gouty arthritis. It could play important role during chronic gouty arthritis treatment. Chronic gouty arthritis by up regulating mir-339-5p, mir-486-5p, mi R-361-5p expressions etc. At the same time, long-term using Chuanhutongbi mixture can effectively prevent and treat chronic gout and reducing the frequency of acute episode, and the side effects was significantly lower than those of allopurinol, which is worthy of clinical promotion.