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
-
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.
-
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。 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。 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。 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。 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。 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。 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。 -
[1] 李杰, 庄辉. 病毒性肝炎流行病学进展[J]. 肝脏, 2012, 17(1): 2-5. https://www.cnki.com.cn/Article/CJFDTOTAL-ZUAN201201004.htm [2] ARAIN SQ, TALPUR FN, CHANNA NA, et al. Serum lipid profile as a marker of liver impairment in hepatitis B cirrhosis patients[J]. Lipids Health Dis, 2017, 16(1): 51. doi: 10.1186/s12944-017-0437-2 [3] 林楠, 黄娇凤, 李建英, 等. 核苷(酸)类似物长期治疗失代偿期乙型肝炎肝硬化的Meta分析[J]. 中国临床研究, 2017, 30(2): 168-174. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGCK201702007.htm [4] CUBERO FJ. Shutting off inflammation: A novel switch on hepatic stellae cells[J]. Hepatology, 2016, 63(11): 1086-1089. http://www.onacademic.com/detail/journal_1000039123469610_efd9.html [5] 中华医学会肝病学分会, 中华医学会感染病学分会. 慢性乙型肝炎防治指南(2015年版)[J]. 实用肝脏病杂志, 2016, 39(1): 1-20. doi: 10.3969/j.issn.1672-5069.2016.01.001 [6] 中国中西医结合学会消化系统疾病专业委员会. 肝硬化中西医结合诊疗共识[J]. 中国中西医结合消化杂志, 2011, 19(4): 277-279. https://www.cnki.com.cn/Article/CJFDTOTAL-ZXPW201104035.htm [7] 中药新药临床研究指导原则[M]. 北京: 中国医药科技出版社, 2002: 85-86. [8] RATHINAM VA, JIANG Z, WAGGONER SN, et al. The AIM2 inflammasome is essential for host defense against cytosolic bacteria and DNA viruses[J]. Nat Immunol, 2010, 11(5): 395-402. doi: 10.1038/ni.1864 [9] INZAUGARAT ME, INZAUGARAT CD, HOLTMANN TM, et al. Nlr family pyrin domain-containing 3 inflammasone activation in hepatic stellate cells induces liver fibrosis in mice[J]. Hepatology, 2019, 69(2): 845-859. doi: 10.1002/hep.30252 [10] WREE A, EGUCHI A, MCGEOUGH MD, et al. NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice[J]. Hepatology, 2014, 59(3): 898-910. doi: 10.1002/hep.26592 [11] 莫志华, 王振常, 黄晶晶, 等. 柔肝化纤颗粒治疗肝硬化肝肾阴虚瘀血阻络证临床观察[J]. 航空航天医学杂志, 2014, 25(2): 246-247. doi: 10.3969/j.issn.2095-1434.2014.02.087 [12] 刘敏, 李羡平. 关幼波治疗肝硬化腹水的经验[J]. 中医药通报, 2006(4): 11-12. doi: 10.3969/j.issn.1671-2749.2006.04.004 [13] 王振常, 柳明, 黄晶晶, 等. 应用模糊集理论对壮肝逐瘀煎抗肝纤维化的优化研究[J]. 中国医药导报, 2013, 10(29): 14-17. doi: 10.3969/j.issn.1673-7210.2013.29.005 [14] 覃伟华, 王振常, 黄晶晶, 等. 柔肝化纤颗粒联合阿德福韦酯治疗慢性乙型肝炎纤维化的临床效果及安全性评价[J]. 检验医学与临床, 2014, 11(11): 1532-1535. doi: 10.3969/j.issn.1672-9455.2014.11.034 [15] 黄瑞华, 王振常, 黄晶晶, 等. 柔肝化纤颗粒联合西药治疗慢性乙肝肝纤维化随机平行对照研究[J]. 实用中医内科杂志, 2014, 28(1): 99-102. https://www.cnki.com.cn/Article/CJFDTOTAL-SYZY201401050.htm [16] 黄瑞华, 王振常, 黄晶晶, 等. 对比分析柔肝化纤颗粒抗肝纤维化前后的指标变化[J]. 航空航天医学杂志, 2014, 25(4): 505-506. doi: 10.3969/j.issn.2095-1434.2014.04.048 [17] 贾新菊, 康岩, 杜丽娜, 等. 黄芪甲苷抑制NLRP3/IL-1β轴改善高糖诱导的血管平滑肌细胞炎症反应[J]. 上海中医药大学学报, 2020, 34(3): 62-69. https://www.cnki.com.cn/Article/CJFDTOTAL-SHZD202003011.htm [18] 孟利娜. 薏苡仁蛋白依赖IKK/NF-κB通道控制炎症及改善2型糖尿病胰岛素抵抗作用[D]. 合肥: 合肥工业大学, 2018. [19] HSU BY, KUO YC, CHEN BH. Polysaccharide isolated from zizyphus jujuba (Hong zao)inhibits interleukin-2 production in jurkat T cells[J]. J Tradit Chin Med, 2014, 4(2): 131-132. [20] 许丹, 林峰, 朱小语, 等. 牡蛎肽对免疫抑制小鼠免疫功能的影响[J]. 北京大学学报(医学版), 2016, 48(3): 392-397. doi: 10.3969/j.issn.1671-167X.2016.03.003 [21] TOSHIHIRO M, MAI M, YU T, et al. Hyaluronidase inhibitors from Takuran, Lycopus lucidus[J]. Chem Pharm Bull, 2010, 58(3): 393-394. http://www.jstage.jst.go.jp/article/cpb/58/3/58_3_394/_pdf [22] TRUONG V, KO SY, JUN M, et al. Quercitrin from toona sinensis(Juss. ) M. Roem. attenuates acetaminophen-induced acute liver toxicity in hepG2 cells and mice through induction of antioxidant machinery and inhibition of inflammation[J]. Nutrients, 2016, 8(7): 430-431. doi: 10.3390/nu8070430 [23] 张目涵, 周力为, 贺欣, 等. IL-27在小鼠结肠炎中的作用及对NLRP3炎症小体的影响[J]. 中国病理生理杂志, 2018, 34(6): 1089-1094. doi: 10.3969/j.issn.1000-4718.2018.06.021
计量
- 文章访问数: 443
- HTML全文浏览量: 80
- PDF下载量: 281
- 被引次数: 0