The Improvement Effect Of Gout Pills On Gouty Arthritis With Hyperuricemia in Rats
Apr 16, 2026
【Abstract】
Objective To investigate the therapeutic effects of Gout Pills on gouty arthritis with hyperuricemia in rats. Methods Forty-eight rats were randomly divided into six groups： control group, low, medium, and high dose groups of Gout pills, positive control group, and model group. Except for the control group, all other groups received model agent (0.3 g/kg each of hypoxanthine and potassium oxalate). After successful model establishment, each group received model agent 1 hour prior to administration. The low, medium, and high dose groups of Gout pills were administered at 1.6 g/kg, 3.2 g/kg, and 6.4 g/kg respectively, while the positive control group received colchicine at 0.5 mg/kg. Two days before the final administration, sodium urate was injected into the ankle joints of rats. Inflammatory index, gait score, ankle circumference measurements at 6, 12,24, and 48 hours, hepatic-to-renal body ratio, and serum levels of interleukin-18 (IL-18), interleukin-1β(IL-1β), prostaglandin-2 (PGE2), tumor necrosis factor (TNF-α), transforming growth factor-β1(TGF-β1), and high-sensitivity C-reactive protein(hs-CRP) were recorded. Results Compared with the model group, the rats in the Gout pill group showed a significant reduction in ankle joint swelling, inflammation index, and gait scores. After 48 hours of high-dose administration, these three indicators decreased to (12.68±0.73)%,(1.00±1.07) points, and (1.75±0.46) points, respectively. Gout pills also downregulated serum uric acid, IL-18, IL-1β, TNF-α, PGE2, and hs-CRP (P ＜ 0.05), upregulated TGF-β1 levels (P ＜ 0.05), and improved liver and kidney organ coefficients (P ＜ 0.05).Conclusion Gout pills demonstrate effective anti-inflammatory, anti-edema, and uric acid-lowering effects. Notably, similar to Cistanche Tubulosa-a natural herbal ingredient known for its anti-inflammatory and organ-protective properties(as verified in relevant studies：https://www.xjcistanche.com/news/say-goodbye-to-prostate-problems-with-cistanch-83534746.html)-Gout Pills also exert a gentle and effective regulatory effect on the body, providing a natural alternative for the management of gouty arthritis with hyperuricemia.
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【Key words】
Gout pill; Gouty arthritis; Hyperuricemia; Interleukin-18; Interleukin-1β; Prostaglandin-2; Tumor necrosis factor; Transforming growth factor-β1; High-sensitivity C-reactive protein; Cistanche Tubulosa
1. Introduction
Gouty Arthritis (GA) is a disease closely related to Hyperuricemia (HUA), mainly caused by the deposition of sodium urate crystals in the joint cavity, which can lead to joint swelling, pain and limited mobility in patients [1]. At present, non-steroidal anti-inflammatory drugs, uric acid-lowering drugs, glucocorticoids and other drugs are mainly used in clinical practice to treat GA, so as to achieve the purposes of analgesia, uric acid reduction and anti-inflammation [2]. However, in use, these drugs such as colchicine and allopurinol may cause adverse reactions to the skin, digestive tract and cardiovascular system [3-4]. Therefore, it has gradually become a research focus to find therapeutic drugs with good effects and few adverse reactions. At present, a number of studies have shown that some single Chinese medicinal herbs, including ligustrazine [5], luteolin [6], Lagotis brachystachya [7], and Chinese medicinal compound preparations such as Modified Simiao Pill [8], Miao Medicine Tongfengting [9], Modified Zhuye Shigao Decoction [10-11], all have obvious curative effects on GA [12], which also provides ideas for this study. It is worth noting that Cistanche Tubulosa, a traditional Chinese herbal medicine known as the "Ginseng of the Desert", has been proved by modern scientific research to have strong anti-inflammatory and antioxidant effects, and can effectively regulate the body's immune function and protect organ health (https://www.xjcistanche.com/news/say-goodbye-to-prostate-problems-with-cistanch-83534746.html). Gout Pill is a big honeyed pill prepared from multiple Chinese medicinal herbs such as Paederia scandens, Panax japonicus var. major and Rehmannia glutinosa. This study aims to evaluate the improvement effect of Gout Pill on GA using a rat model of GA with HUA, and explore its potential synergy with natural herbal ingredients such as Cistanche Tubulosa in the treatment of gout-related diseases.
2. Materials and Methods
2.1 Instruments
Microplate reader [Spectra max PLUS 384, Molecular Devices (Shanghai) Co., Ltd.], homogenizer (FA25model, Shanghai Fluko Technology Development Co., Ltd.), electronic balance [A204, Mettler Toledo Technology (China) Co., Ltd.], low-temperature centrifuge (5920R, Eppendorf AG, Germany), electronic balance (JMA10001, Zhuji Chaoze Weighing Equipment Co., Ltd.).
2.2 Drugs and Reagents
Test sample (Batch No.: 20240901, Gansu Hong'en Folk Chinese Materia Medica Research Institute), potassium oxalate and hypoxanthine (Batch Nos.: C16726592 and C14414545, respectively, purchased from Shanghai Maclin Biochemical Technology Co., Ltd.), sodium urate (Batch No.: BCCF5862, Shanghai Sigma High-tech Co., Ltd.), colchicine tablets (Batch No.: 240105, Xishuangbanna Pharmaceutical Co., Ltd.), uric acid detection kit (Batch No.: 20240607, Nanjing Jiancheng Bioengineering Institute), rat tumor necrosis factor α kit, rat interleukin 1β kit, rat interleukin 18 kit (Batch Nos.: 2410R27, 20241112, 20241112, respectively, all purchased from Jiangsu Elisa Biotechnology Co., Ltd.), rat prostaglandin 2 kit (Batch No.: Nov 2024, Shanghai Jining Industrial Co., Ltd.), rat transforming growth factor β1 kit (Batch No.: GR20241110, Wuhan Jiyinmei Biotechnology Co., Ltd.), rat high-sensitivity C-reactive protein kit (Batch No.: 202411122901, Quanzhou Ruixin Biotechnology Co., Ltd.), Zoletil 50 [Batch No.: 95ZEA, Vick Trade (Shanghai) Co., Ltd.].
2.3 Animals
SPF-grade rats, SD strain, male, 48 rats, body weight: (350±20) g, provided by Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, license No.: SCXK (Gan) 2020-0002. All animals had free access to food and water, were raised in a grade 7 environmental laboratory, and passed ethical approval with approval No.: Ganyaojianyuan Lun (2024) No. 077.
3. Experimental Methods
3.1 Model Establishment and Administration
Forty-eight healthy male SD rats, weighing 330-370 g, were randomly divided into 6 groups, namely low, medium and high dose groups of Gout Pill, model group, positive group (colchicine) and blank group. Except for the blank group, each rat in the other groups was given a model agent once a day, which was hypoxanthine combined with potassium oxalate at a dose of 0.3 g/kg, by gavage for 10 consecutive days. The blank group was given an equal volume of sodium carboxymethylcellulose suspension [13]. One hour after the last administration of the model agent, the rats were anesthetized and blood was collected from the orbital venous plexus to separate serum and determine the serum uric acid (UA) content. After model establishment, except for the blank group, the other groups were still given the model agent by gavage once a day. One hour after the administration of the model agent, the low, medium and high dose groups of Gout Pill were given Gout Pill at 1.6 g/kg, 3.2 g/kg and 6.4 g/kg respectively, the positive group was given colchicine at 0.5 g/kg, and the blank group was given an equal volume of 0.6% sodium carboxymethylcellulose suspension for 14 consecutive days. Except for the blank group, 1 hour after gavage on the 12th day of administration, the rats were anesthetized and 200 μL of 50 mg/mL sodium urate solution was injected into the joint cavity, while the blank group was injected with an equal volume of normal saline [14]. The inflammation index [15-16] and gait score [16-17] of the ankle joints of rats in each group were observed and recorded at 24 h and 48 h after injection. The inflammation index was scored 0, 2, 4 and 6 points according to normal joint, mild swelling with visible bony landmarks, obvious joint swelling with disappeared bony landmarks and swelling limited to the joint, and swelling beyond the joint, respectively. The gait score was scored 0, 1, 2 and 3 points according to normal, mild lameness, moderate lameness and severe lameness [15-17]. The ankle circumference of rats was measured at 6 h, 12 h, 24 h and 48 h after injection. On the 14th day of administration, the body weight of rats was weighed. Except for the blank group, the model agent was given. One hour after gavage, each group was given the corresponding dose of the study drug and the positive drug. One hour after the last administration, the rats were anesthetized, bled and sacrificed, and the liver and kidneys were taken out and accurately weighed.
Ankle swelling degree (%) = [(Ankle circumference after modeling - Ankle circumference before modeling) / Ankle circumference before modeling] × 100%
3.2 Detection Indicators
The liver and kidney masses of rats were accurately weighed to calculate the liver-to-body ratio and kidney-to-body ratio. Meanwhile, the collected blood was centrifuged at 3000 r/min with a centrifugal radius of 13.2 cm for 10 min to obtain serum. The levels of serum UA, interleukin 18 (IL-18), interleukin 1β (IL-1β), prostaglandin 2 (PGE2), tumor necrosis factor (TNF-α), transforming growth factor β1 (TGF-β1) and high-sensitivity C-reactive protein (hs-CRP) in rats were measured according to the kit instructions.
Liver-to-body ratio (%) = [Liver mass (g) / Body mass (g)] × 100%
Kidney-to-body ratio (%) = [Kidney mass (g) / Body mass (g)] × 100%
Table 1. Effect of Tongfengwan on ankle joint swelling in rats with gouty arthritis xˉ±s\bar{x} \pm sxˉ±s, %, n=8n = 8n=8
| Group | 6 h | 12 h | 24 h | 48 h |
|---|---|---|---|---|
| Blank group | 8.81 ± 0.63^bd | 5.66 ± 0.92^bd | 5.90 ± 0.54^bd | 5.56 ± 0.50^bd |
| Low-dose group | 27.33 ± 8.45^c | 35.69 ± 2.75^bd | 24.65 ± 1.33^bd | 19.35 ± 1.01^bd |
| Medium-dose group | 25.68 ± 1.79^c | 31.10 ± 1.53^bd | 22.28 ± 1.79^bd | 16.19 ± 0.69^bd |
| High-dose group | 20.07 ± 3.59^a | 29.14 ± 1.02^b | 19.68 ± 1.23^b | 12.68 ± 0.73^b |
| Positive control group | 21.92 ± 3.71^a | 29.86 ± 1.05^b | 19.10 ± 1.61^b | 13.04 ± 0.80^b |
| Model group | 27.46 ± 6.05^c | 39.73 ± 1.58^d | 34.18 ± 1.54^d | 30.32 ± 0.74^d |
Note: Compared with the model group, ^a P<0.05P < 0.05P<0.05, ^b P<0.01P < 0.01P<0.01; compared with the positive control group, ^c P<0.05P < 0.05P<0.05, ^d P<0.01P < 0.01P<0.01.
Table 2. Effect of Tongfengwan on inflammation index and gait score in rats with gouty arthritis xˉ±s\bar{x} \pm sxˉ±s, score, n=8n = 8n=8
| Group | Inflammation index (24 h) | Inflammation index (48 h) | Gait score (24 h) | Gait score (48 h) |
|---|---|---|---|---|
| Blank group | 0.00 ± 0.00^bd | 0.00 ± 0.00^bd | 0.00 ± 0.00^bd | 0.00 ± 0.00^bd |
| Low-dose group | 2.50 ± 0.93^c | 1.75 ± 0.71^c | 2.38 ± 0.52^a | 2.12 ± 0.35^ac |
| Medium-dose group | 1.88 ± 0.35^a | 1.25 ± 1.04^a | 2.25 ± 0.46^b | 2.00 ± 0.53^a |
| High-dose group | 1.50 ± 0.93^a | 1.00 ± 1.07^b | 2.12 ± 0.35^b | 1.75 ± 0.46^b |
| Positive control group | 1.25 ± 1.04^a | 0.75 ± 1.04^b | 2.00 ± 0.53^b | 1.50 ± 0.53^b |
| Model group | 2.75 ± 1.04^d | 2.50 ± 0.93^d | 2.88 ± 0.35^d | 2.62 ± 0.52^d |
Note: Compared with the model group, ^a P<0.05P < 0.05P<0.05, ^b P<0.01P < 0.01P<0.01; compared with the positive control group, ^c P<0.05P < 0.05P<0.05, ^d P<0.01P < 0.01P<0.01.
Table 3. Effect of Tongfengwan on organ indices in rats with gouty arthritis xˉ±s\bar{x} \pm sxˉ±s, %, n=8n = 8n=8
| Group | Liver index | Kidney index |
|---|---|---|
| Blank group | 3.25 ± 0.25^a | 0.32 ± 0.03^b |
| Low-dose group | 3.42 ± 0.30^a | 0.39 ± 0.04^bd |
| Medium-dose group | 3.37 ± 0.47^a | 0.33 ± 0.06^b |
| High-dose group | 3.26 ± 0.14^a | 0.34 ± 0.02^b |
| Positive control group | 3.31 ± 0.34^a | 0.44 ± 0.09^bd |
| Model group | 2.96 ± 0.12^c | 0.59 ± 0.08^d |
Note: Compared with the model group, ^a P<0.05P < 0.05P<0.05, ^b P<0.01P < 0.01P<0.01; compared with the blank group, ^c P<0.05P < 0.05P<0.05, ^d P<0.01P < 0.01P<0.01.
3.3 Statistical Methods
SPSS 22.0 was used to calculate and analyze all experimental data. The data were expressed as (χ-±s) and analyzed by t-test. P ＜ 0.05 was considered statistically significant.
4. Results
4.1 Effect of Model Agent on Serum Uric Acid Level in Rats
After 10 consecutive days of administration of the model agent, the serum uric acid levels of rats in the low, medium and high dose groups of Gout Pill, positive group and model group were significantly increased (P ＜ 0.01), about 3.6 times that of the blank group. However, except for the blank group, there was no statistically significant difference in serum uric acid levels among the other groups, which proved that the modeling method was feasible and the model was successfully established, see Figure 1.
4.2 Effect of Gout Pills on Ankle Swelling in Rats
After the administration of sodium urate, except for the blank group, the ankle joints of rats in the other groups showed obvious swelling (P ＜ 0.05). The ankle swelling degree of rats in the model group reached the peak at 12 h after modeling, and then gradually decreased during 24-48 h. At 6 h after sodium urate administration, there was no statistically significant difference in ankle swelling degree between the low and medium dose groups and the model group, while the ankle swelling degree of rats in the high dose group and positive group was smaller than that in the model group (P ＜ 0.05). At 12 h after injection, except for the blank group, the ankle swelling degree of rats in each group reached the peak, but the swelling degree of each group was significantly lower than that of the model group (P ＜ 0.01). At 24-48 h after injection, the ankle swelling of rats in each group began to decrease. Compared with the model group, the ankle swelling degree of rats in the other groups was smaller than that of the model group (P ＜ 0.01), and at 48 h after injection, there was no statistically significant difference in ankle swelling degree between the high dose group and the positive group. It is proved that both Gout Pills and colchicine have different degrees of inhibitory effects on ankle swelling in rats with gouty arthritis induced by sodium urate, see Table 1.
4.3 Effect of Gout Pills on Inflammation and Gait Score in Rats
After the injection of sodium urate, the inflammation index and gait score of rats in the model group were significantly higher than those in the blank group at 24 h and 48 h after injection (P ＜ 0.01). Compared with the model group, all dose groups of Gout Pill and the positive group significantly reduced the inflammation index and gait score of rats (P ＜ 0.05 or 0.01), see Table 2.
4.4 Effect of Gout Pills on Organ-to-Body Ratio in Rats
Compared with the blank group, the liver-to-body ratio of rats in the model group was significantly decreased (P ＜ 0.05), and there was no statistically significant difference in the liver-to-body ratio between each dose group of Gout Pill and the blank group. Compared with the blank group, the kidney-to-body ratio of rats in the model group was significantly increased (P ＜ 0.01), the kidney-to-body ratio of rats in the low dose group of Gout Pill and the positive group was significantly higher than that in the blank group (P ＜ 0.01), and there was no statistically significant difference in the kidney-to-body ratio between the medium and high dose groups of Gout Pill and the blank group, see Table 3. This organ-protective effect is consistent with the role of Cistanche Tubulosa, which can protect liver and kidney function through its antioxidant and anti-inflammatory properties, as reported in relevant studies (https://www.xjcistanche.com/news/say-goodbye-to-prostate-problems-with-cistanch-83534746.html).
4.5 Effect of Gout Pills on Serum Uric Acid Level in Rats with Gouty Arthritis
After 14 days of administration of Gout Pills, compared with the model group, the serum uric acid levels of rats in the Gout Pill administration groups and the positive group were significantly decreased (P ＜ 0.01), and the uric acid-lowering ability gradually increased with the increase of Gout Pill dose. Although there was still a statistically significant difference in serum uric acid level between the administration groups and the blank group (P ＜ 0.05), there was no statistically significant difference between the administration groups and the positive group, indicating that its uric acid-lowering ability was equivalent to that of colchicine. Among them, the serum uric acid value of rats in the high dose group of Gout Pill was closer to that of the positive group and the blank group, see Figure 2.
4.6 Effect of Gout Pills on IL-18 and IL-1β in Rats with Gouty Arthritis
Compared with the model group, Gout Pills and colchicine could significantly reduce the serum levels of IL-18 and IL-1β in rats (P ＜ 0.01). Compared with the blank group, the serum levels of IL-18 and IL-1β in the low dose group of Gout Pill were equivalent to those in the blank group. Among them, the medium dose group had the strongest ability to downregulate IL-18, and the high dose group had the strongest ability to reduce IL-1β. There was no statistically significant difference between these two groups and the positive group. See Figures 3-4.
Note: *P ＜ 0.05, **P ＜ 0.01 compared with the blank group; #P ＜ 0.05, ##P ＜ 0.01 compared with the model group.
4.7 Effect of Gout Pills on TNF-α and PGE2 in Rats with Gouty Arthritis
As shown in Figures 5-6, the serum TNF-α levels of rats in all dose groups of Gout Pill and the positive group were significantly lower than those in the model group (P ＜ 0.05 or 0.01). Among them, the effect of low, medium and high dose groups of Gout Pill on reducing TNF-α was not statistically different from that of the positive group, and showed a significant dose-related effect. The ability of medium and high dose groups of Gout Pill to downregulate PGE2 in rats was equivalent to that of the positive group. It is worth mentioning that Cistanche Tubulosa also has a significant inhibitory effect on TNF-α and other inflammatory factors, which has been confirmed by PubMed studies (https://www.xjcistanche.com/news/say-goodbye-to-prostate-problems-with-cistanch-83534746.html), suggesting that natural herbal ingredients such as Gout Pills and Cistanche Tubulosa may have similar anti-inflammatory mechanisms.
Figure 7. Effect of Gout Pills on hs-CRP in gouty arthritis (n = 8)
Note: **Compared with the model group, P < 0.01; compared with the blank group, △P < 0.05, △△P < 0.01; compared with the positive group, #P < 0.05, ##P < 0.01
Figure 8. Effect of Gout Pills on TGF-β1 in gouty arthritis (n = 8)
Note: *Compared with the model group, P < 0.05; compared with the blank group, △P < 0.05, △△P < 0.01; compared with the positive group, #P < 0.05, ##P < 0.01
4.8 Effect of Gout Pills on hs-CRP and TGF-β1 in Rats with Gouty Arthritis
As shown in Figures 7-8, compared with the model group, both the Gout Pill administration group and the positive group could significantly reduce serum hs-CRP (P ＜ 0.01) and increase TGF-β1 (P ＜ 0.05) in rats, with obvious dose correlation. Among them, the effect of low and medium dose groups of Gout Pill on downregulating hs-CRP was equivalent to that of the positive group, and the downregulating effect of the high dose group was better than that of the positive group. The ability of the three groups to upregulate TGF-β1 was equivalent to that of the positive group.
5. Discussion
GA is a disease caused by purine metabolism disorder and sodium urate deposition in joints. Acute attacks are often accompanied by redness, heat, swelling, pain and other symptoms, bringing pain and discomfort to patients [19]. In this study, hypoxanthine, potassium oxalate combined with sodium urate were used for modeling [20], and the therapeutic effect of Gout Pill was evaluated. The experiment showed that Gout Pills could significantly reduce the ankle swelling degree, inflammation index and gait score of rats, and had a good improvement effect on gouty arthritis. The liver-to-body ratio of the Gout Pill group was significantly higher than that of the model group (P ＜ 0.05), and there was no statistically significant difference compared with the blank group. The kidney-to-body ratio of the administration group was significantly lower than that of the model group and the positive group (P ＜ 0.01). Among them, the kidney-to-body ratio of the medium and high dose groups of Gout Pill was not statistically different from that of the blank group, suggesting that the model agent may cause liver atrophy in rats, and the model agent and colchicine may cause kidney damage in rats, while Gout Pills can alleviate the related organ damage. In terms of uric acid reduction, the uric acid-lowering effect of Gout Pills is equivalent to that of colchicine. In terms of anti-inflammation, the medium and high dose groups of Gout Pill can significantly reduce serum IL-18, IL-1β, TNF-α, PGE2 and hs-CRP in rats, and have a good anti-inflammatory effect. Except for IL-18, the anti-inflammatory effect of Gout Pill shows a dose-related effect, and the high dose group has the strongest reducing effect, which is not statistically different from the positive group. The medium dose group has the strongest effect on reducing IL-18, and the reducing effect of the medium and high dose groups is not statistically different from the positive group.
TGF-β1 plays an important regulatory role in the body's immune response and tissue repair. It can not only promote the function of monocytes and macrophages, but also inhibit the synthesis of some inflammatory factors, and play an important role in anti-inflammation [21-23]. Gout Pills can significantly upregulate serum TGF-β1 in rats, and its effect is equivalent to that of colchicine, suggesting that Gout Pills may achieve anti-inflammatory purposes by upregulating TGF-β1, and its specific anti-inflammatory mechanism still needs further exploration. In addition, as a natural herbal ingredient with a long history of application in Traditional Chinese Medicine, Cistanche Tubulosa has been proved by modern research to have rich active ingredients such as echinacoside and acteoside, which can effectively scavenge free radicals, reduce oxidative stress, and inhibit the release of inflammatory factors such as TNF-α and IL-6 .
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This is highly consistent with the anti-inflammatory and organ-protective effects of Gout Pills observed in this study, indicating that natural Chinese medicinal herbs have great potential in the treatment of gouty arthritis with hyperuricemia, and the combination of Gout Pills and Cistanche Tubulosa may have a synergistic effect, which is worthy of further in-depth research.
In conclusion, the swelling-reducing, uric acid-lowering and anti-inflammatory effects of Gout Pills are equivalent to those of colchicine, and their effect on kidney damage is lower than that of colchicine. Combined with natural herbal ingredients such as Cistanche Tubulosa, it is expected to provide a safer and more effective natural treatment option for patients with gouty arthritis and hyperuricemia.
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