Effect of Rougan Huaxian Granules on the Expression of NLRP3 Inflammasomes and Related-Products in Peripheral Blood of Patients with Liver and Kidney Yin Deficiency Type of Compensated Hepatitis B Cirrhosis
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摘要: 目的 观察柔肝化纤颗粒对代偿期乙肝肝硬化肝肾阴虚型患者的临床疗效，及对血清核苷酸寡聚化结构域样受体蛋白3(NLRP3)炎症小体及其产物表达水平的影响。方法 80例患者随机分为治疗组、对照组各40例。2组按指南给予恩替卡韦口服，治疗组加服柔肝化纤颗粒。2组疗程均为24周。疗程结束后，比较2组肝功能、肝纤四项[Ⅲ型前胶原(PC-Ⅲ)、Ⅳ型胶原(Ⅳ-C)、层黏蛋白(LN)、透明质酸(HA)]、肝脾影像学指标、乙肝病毒(HBV)-DNA、中医证候积分、临床疗效、血清白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)、肿瘤坏死因子-α(TNF-α)蛋白表达水平及NLRP3炎症小体、半胱氨酸天冬氨酸蛋白酶1(Caspase-1)、IL-1β、IL-18 mRNA的表达。结果 治疗后2组中医证候积分均较治疗前降低(P < 0.01), 治疗组优于对照组(P < 0.01)；2组肝功能、肝纤四项、证候积分、肝脾影像学指标均优于治疗前(P < 0.01)，且治疗组各项指标优于对照组(P < 0.01)；2组血清IL-1β、IL-18、TNF-α水平均较治疗前下降，NLRP3、Caspase-1、IL-1β、IL-18 mRNA水平优于治疗前(P < 0.01)，治疗组均较对照组作用显著(P < 0.01)；治疗组总有效率高于对照组(P < 0.05)。结论 柔肝化纤颗粒联合恩替卡韦治疗代偿期乙肝肝硬化肝肾阴虚型患者临床疗效显著，其机制可能是通过抑制NLRP3炎症小体、Caspase-1、IL-1β、IL-18及TNF-α表达水平，有效抑制机体炎症反应，从而改善患者临床症状。Abstract: OBJECTIVE To observe the clinical efficacy of Rougan Huaxian granules on patients with liver and kidney-yin deficiency type of compensated hepatitis B cirrhosis and the effects on the expression levels of serum NLRP3 inflammasomes and related-products.METHODS 80 patients were randomly divided into the treatment group and the control group, 40 patients for each. The two groups were given Entacvir orally according to the guideline, while the treatment group was given Rougan Huaqian granules in addition to Entacvir. The course lasted 24 weeks. At the end of the treatment course, the liver function, four indicators of liver fibrosis [procollagen type Ⅲ(PC-Ⅲ), collagen type Ⅳ (Ⅳ-C), laminin (LN), hyaluronic acid (HA)], liver and spleen imaging indexes, HBV DNA, traditional Chinese medicine (TCM) syndrome scores, clinical efficacy, serum protein levels of interleukin-1β (IL-1β), interleukin-18 (IL-18), and tumor necrosis factor-α (TNF-α), serum levels of NLRP3 inflammasomes, cysteine aspartate protease 1 (Caspase-1), IL-1β, IL-18 mRNA were compared between the two groups.RESULTS After treatment, the TCM syndrome scores of both groups were lower than those before treatment (P < 0.01), and the scores of the treatment group were better than those of the control group (P < 0.01). The liver function, four indicators of liver fibrosis, TCM syndrome scores, liver and spleen imaging indexes of both groups were better than those before treatment (P < 0.01). The indexes of the treatment group were better than those of the control group (P < 0.01). The serum protein levels of IL-1β, IL-18, TNF-α in both groups were lower than those before treatment. The levels of NLRP3, Caspase-1, IL-1β, and IL-18 mRNA were better than those before treatment (P < 0.01), and the treatment group was significantly better than the control group (P < 0.01). The total effective rate of the treatment group was 87.5%, higher than the control group (P < 0.05).CONCLUSION The clinical efficacy of Rougan Huaxian granules combined with Entacvir is significantly improved in the treatment of patients with liver and kidney yin deficiency type of compensated hepatitis B cirrhosis. The mechanisms may involve inhibiting the expression levels of NLRP3 inflammasomes, Caspase-1, IL-1β, IL-18, and TNF-α, effectively controlling the inflammatory response of the body, thus improving the clinical symptoms of patients.
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表 1 荧光定量PCR实验所用引物序列
引物名 引物序列(5'-3') Caspase-1 F: ACGCCTTGCCCTCATAAT
R: TCTAATACATCTGGGACTTCTTIL-1β F: ATGCACCTGTACGATCACTG
R: ACAAAGGACATGGAGAACACCIL-18 F: CATTGACCAAGGAAATCGGC
R: CACAGAGATAGTTACAGCCATACCNLRP3 F: GTGTTTCGAATCCCACTGTG
R: TCTGCTTCTCACGTACTTTCTGGAPDH F: AGACAGCCGCATCTTCTTGT
R: CTTGCCGTGGGTAGAGTCAT
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表 2 2组患者治疗前后中医证候积分比较( x±s，n=40)
组别 时间 胁肋隐痛 腰膝酸软 五心烦热 头昏目眩 两目干涩 失眠多梦 治疗组 治疗前 2.65±0.27 2.45±0.41 2.44±0.43 2.12±0.45 2.09±0.56 2.76±0.71 治疗后 0.98±0.09**## 0.31±0.10**## 0.44±0.21**## 0.34±0.20**## 0.76±0.30**## 0.98±0.51**## 对照组 治疗前 2.57±2.24 2.34±0.44 2.66±0.44 2.13±0.49 2.19±0.57 2.89±0.50 治疗后 1.53±0.52** 0.82±0.14** 1.23±0.29** 1.23±0.31** 1.59±0.45** 1.87±0.38** 注：与同组治疗前比较，**P＜0.01；与对照组同期比较，##P＜0.01。
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表 3 2组患者治疗前后肝功能指标比较( x±s，n=40)
组别 时间 ALT/(U·L-1) AST/(U·L-1) TBIL/(μmol·L-1) ALB/(g·L-1) 治疗组 治疗前 65.92±18.78 66.02±18.88 41.57±4.98 25.99±2.17 治疗后 47.01±4.96**## 46.79±5.03**## 18.04±2.14**## 40.43±3.65**## 对照组 治疗前 69.10±19.40 70.19±18.28 40.72±4.88 25.24±1.79 治疗后 52.68±8.78** 54.94±9.37** 26.94±2.97** 34.93±3.03** 注：与同组治疗前比较，**P＜0.01；与对照组同期比较，##P＜0.01。
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表 4 2组患者治疗前后肝纤四项水平比较( x±s，ng·mL-1, n=40)
组别 时间 PC-Ⅲ Ⅳ-C LN HA 治疗组 治疗前 157.02±68.09 145.51±47.31 235.12±47.19 202.49±92.81 治疗后 110.93±55.55**## 96.01±21.19**## 184.13±14.77**## 101.67±85.97**## 对照组 治疗前 165.01±71.98 153.17±49.95 231.71±45.82 197.26±85.97 治疗后 124.51±50.48** 113.27±27.48** 204.08±35.55** 153.23±53.85** 注：与同组治疗前比较，**P＜0.01；与对照组同期比较，##P＜0.01。
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表 5 2组患者治疗前后脾厚度、Dpv、Dsv及血清HBV-DNA水平比较( x±s, n=40)
组别 时间 HBV-DNA/ (×104U·mL-1) 脾厚度/mm Dsv/mm Dpv/mm 治疗组 治疗前 5.24±0.43 43.71±3.67 10.98±1.02 15.13±1.31 治疗后 ＜0.01** 36.62±2.48**## 8.02±0.67**## 10.96±0.91**## 对照组 治疗前 5.28±0.35 45.73±3.88 10.86±1.01 15.01±1.42 治疗后 ＜0.01** 40.49±3.62** 9.12±0.85** 12.54±1.10** 注：与同组治疗前比较，**P＜0.01；与对照组同期比较，##P＜0.01。
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表 6 2组患者治疗前后血清IL-1β、IL-18及TNF-α水平比较( x±s，n=40)
组别 时间 TNF-α/(pg·mL-1) IL-1β/(ng·L -1) IL-18/(ng·L -1) 治疗组 治疗前 33.04±8.25 32.31±9.34 26.71±3.58 治疗后 21.99±8.08**## 19.34±7.33**## 20.54±2.28**## 对照组 治疗前 51.92±8.74 31.21±8.34 42.38±5.79 治疗后 36.86±6.29** 24.35±7.61** 29.77±4.20** 注：与同组治疗前比较，**P＜0.01；与对照组同期比较，##P＜0.01。
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表 7 2组患者治疗前后NLRP3、Caspase-1、IL-1β及IL-18 mRNA表达比较( x±s，n=40)
组别 时间 NLRP3 Caspase-1 IL-1β IL-18 治疗组 治疗前 2.69±1.56 1.35±0.28 5.87±2.12 6.37±2.86 治疗后 1.95±0.78**## 0.61±0.17**## 3.33±1.41**## 4.21±1.17**## 对照组 治疗前 2.70±1.59 1.34±0.31 5.88±2.15 6.38±2.89 治疗后 2.34±0.98** 0.98±0.18** 4.78±1.47** 5.18±2.21** 注：与同组治疗前比较，**P＜0.01；与对照组同期比较，##P＜0.01。
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表 8 2组患者临床疗效比较(n=40)
组别 治愈 显效 有效 无效 总有效率/% 治疗组 20 9 6 5 87.50* 对照组 15 7 5 13 67.50 注：与对照组比较, χ2=4.59，*P＜0.05。
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