Efficacy and safety of Salvia miltiorrhiza (Salvia miltiorrhiza Bunge) and ligustrazine injection in the adjuvant treatment of early-stage diabetic kidney disease: A systematic review and meta-analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Salvia miltiorrhiza and ligustrazine injection is a compound injection composed of the extract from Salvia miltiorrhiza and Ligusticum striatum (Ligusticum striatum DC.), has been frequently used for the adjuvant treatment of early-stage diabetic kidney disease (DKD) in China. Safety and efficacy studies in terms of evidence-based medical practice have become more prevalent in application to Chinese Herbal Medicine. It is necessary to assess the efficacy and safety of Salvia miltiorrhiza and ligustrazine injection in the adjuvant treatment of early-stage diabetic kidney disease by conducting a systematic review and meta-analysis of available clinical data. AIM OF THE STUDY: The aim of this systematic review and meta-analysis was to evaluate the efficacy and safety of Salvia miltiorrhiza and ligustrazine injection as an adjunctive therapy to conventional therapies for early-stage DKD. MATERIALS AND METHODS: The Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) checklist was used to structure this study. We searched the English databases PubMed, Cochrane library, and Chinese databases including Chinese journal full text database (CNKI), China Biomedical Documentation Service System (SinoMed), Wanfang digital periodical full text database and Chinese Scientific Journal Database (VIP). Relevant studies were selected based on the inclusion and exclusion criteria. Meta-analysis was performed with RevMan 5.3 software after data extraction and the quality of studies assessment. The quality of the evidence was assessed with the Cochrane risk of bias tool. Sensitivity analysis and Egger's test were performed using Stata 15.0 software. RESULTS: A total of 22 trials were included with 1939 patients. Meta-analysis showed that compared with the control group of conventional western medicine alone, Salvia miltiorrhiza and ligustrazine injection combined with conventional western medicine can achieve better efficacy in the treatment of early-stage DKD, reduce urinary albumin excretion rate (12RCTs, 1181 participants; SMD = -1.82, 95% CI [-2.62, -1.01], P < 0.00001), serum creatinine (13RCTs, 1228 participants; MD = -13.21 µmol/L, 95% CI [-19.58, -6.83], P < 0.0001), ß2-microglobulin (9RCTs, 669 participants; SMD = -1.45, 95% CI [-2.43, -0.48], P = 0.003) and reduce interleukin-6 (4RCTs, 331 participants; MD = -6.38 ng/L, 95% CI [-9.03, -3.78], P < 0.00001), interleukin-18 (2RCTs, 177 participants; MD = -29.78 ng/L, 95% CI [-41.51, -18.05], P < 0.00001), tumor necrosis factor-α (4RCTs, 331 participants; MD = -18.03 ng/L, 95% CI [-22.96, -13.09], P < 0.00001), with statistical differences and alleviate the body inflammatory response effectively. CONCLUSION: Based on the existing evidence, that Salvia miltiorrhiza and ligustrazine injection in the adjuvant treatment of early-stage diabetic kidney disease is safe and effective. However, due to the limitation of the quality of the included studies, the above conclusions need to be further verified by more relevant randomized controlled trials with high-quality large samples.

Anti-fibrotic effects of Salvia miltiorrhiza and Ligustrazine Injection on LX-2 cells involved with increased N-myc downstream-regulated gene 2 expression.

OBJECTIVE: To investigate the effects of Salvia miltiorrhiza and Ligustrazine Injection (SML) on proliferation and apoptosis of human hepatic stellate cell LX-2 and the expression of N-myc downstreamregulated gene 2 (NDRG2, a tumor suppressor gene). METHODS: HSCs from the LX-2 cell line were cultured in vitro. The proliferative state of different initial LX-2 cell numbers was measured using a 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. LX-2 cells were plated in 96-well plates at an approximate density of 2.50×104 cells/mL and cultured for 24 h followed by the application of different concentrations of SML (1, 2, 4 and 8 µL/mL). Cell proliferation was measured using the MTT assay at 24 and 48 h. Apoptosis was detected by flow cytometry at 24 h. LX-2 cells were treated with different concentrations of SML and extracted with protein lysis buffer. The levels of NDRG2 and ß-catenin were measured by Western blot. RESULTS: With the exception of the 1 and 2 µL/mL concentrations, 4 and 8 µL/mL SML inhibited cell proliferation in a concentration-dependent manner at 24 and 48 h (P<0.05). With the exception of the 1 and 2 µL/mL concentrations, the NDRG2 expression level was greatly increased in a concentration-dependent manner. However, the level of ß-catenin was unaffected. CONCLUSION: SML inhibit LX-2 cell proliferation in a concentration-dependent manner, and the mechanism may be associated with NDRG2 over-expression.

Anti-fibrotic effects of Salvia miltiorrhiza and Ligustrazine Injection on LX-2 cells involved with increased N-myc downstream-regulated gene 2 expression / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of Salvia miltiorrhiza and Ligustrazine Injection (SML) on proliferation and apoptosis of human hepatic stellate cell LX-2 and the expression of N-myc downstreamregulated gene 2 (NDRG2, a tumor suppressor gene).</p><p><b>METHODS</b>HSCs from the LX-2 cell line were cultured in vitro. The proliferative state of different initial LX-2 cell numbers was measured using a 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. LX-2 cells were plated in 96-well plates at an approximate density of 2.50×10cells/mL and cultured for 24 h followed by the application of different concentrations of SML (1, 2, 4 and 8 μL/mL). Cell proliferation was measured using the MTT assay at 24 and 48 h. Apoptosis was detected by flow cytometry at 24 h. LX-2 cells were treated with different concentrations of SML and extracted with protein lysis buffer. The levels of NDRG2 and β-catenin were measured by Western blot.</p><p><b>RESULTS</b>With the exception of the 1 and 2 μL/mL concentrations, 4 and 8 μL/mL SML inhibited cell proliferation in a concentration-dependent manner at 24 and 48 h (P<0.05). With the exception of the 1 and 2 μL/mL concentrations, the NDRG2 expression level was greatly increased in a concentration-dependent manner. However, the level of β-catenin was unaffected.</p><p><b>CONCLUSION</b>SML inhibit LX-2 cell proliferation in a concentration-dependent manner, and the mechanism may be associated with NDRG2 over-expression.</p>