Qiong-Yu-Gao Protects Against Cisplatin-Induced Nephrotoxicity Without Reduce The Anti-Tumor Effect In Mice

Background:Cisplatin, cell cycle non-specific drug, can inhibit cancer cell DNA replication process, has broad-spectrum anti-cancer effect, often used in a variety of solid tumors in clinical. Cisplatin has side effects, including kidney toxicity which is the most common and severe toxicity reaction. At present, research against cisplatin renal toxicity is more and more, but so far no one can solve cisplatin renal toxicity with no affect its curative effect. Qiong-Yu-Gao(QYG) is a classic agent from Song Dynasty, with the function of nourishing yin to strong water, replenishing qi to invigorate the spleen and other effects. It has effects of moistening lung for arresting cough, gastrointestinal conditioning, anti-aging effects, related to the mechanism of anti-inflammatory, antioxidant effects. QYG has been reported can reduce cisplatin renal toxicity when combination with cisplatin. In view of this, QYG could have certain therapeutic potential in the nephrotoxicity of cisplatin.Objective:This research mainly aims to evaluate the effect of QYG against cisplatin renal toxicity through the in vivo experiment, and to explore its preliminary mechanism.Methods:(1) Using liquid chromatography-mass spectrometry(LC-MS) to study the quality control of QYG, detect the composition and their content in QYG. (2) C57BL / 6 male mice were randomly divided into blank group (Control) and Cisplatin group, QYG + Cisplatin group. Mice of control and Cisplatin groups with physiological saline lavage (0.2 ml/day),10 days. QYG+Cisplatin (14 g/kg QYG,0.2 ml/day) 10 days, at day 7, intraperitoneal injection with a single dose of Cisplatin (20 mg/kg) on Cisplatin group and QYG+Cisplatin groups. Three days after injection of cisplatin, take blood from eye, collect both sides of the kidney in mice. According to the creatinine and urea nitrogen detection kit and H&E staining method to evaluate the type of renal function. (3) Using inductively coupled luminous mass spectrometry and immunohistochemical method to detect Pt+ content in mice tissues and urine. The content of Ctrl and OCT2 to evaluate the type of cisplatin transfer function. Using TUNEL staining, immunohistochemical method, the Western bloting assay and qPCR assay to detect cell apoptosis and proliferation, mice kidney tissue inflammation reaction was evaluated by cox-2, IL-1β, TNF alpha mRAN level. (4) Establish a tumor-burdened Lewis Model in mice, the mice were randomly divided into blank group (Control), Model group, Cisplatin, QYG and combined treatment group (Cisplatin+QYG). Control group lavage daily with normal saline, each mouse lavage daily 0.2 ml, model group, the daily physiological saline lavage,0.2 ml per mouse lavage daily, QYG group: QYG lavage, daily dose of 14 g/kg,0.2 ml per mouse lavage daily, cisplatin group:intraperitoneal injection of cisplatin solution, dose of 2 mg/kg, on time, a total of 3 times, cisplatin+ QYG group:intraperitoneal injection of cisplatin solution, dose of 2 mg/kg, day time, a total of 3 times; At the same time daily with QYG lavage,14 g/kg, how much dose each mouse lavage daily 0.2 ml. To measure and record a tumor-burdened mice tumor volume dead mice to evaluate QYG on the effects of a tumor-burdened tumor growth and survival time of mice.Results:21 compounds of QYG were detected using LC-MS, at the same time, the content of 13 of these compounds were showed. Body weight of mice were reduced after cisplatin administrated, serum urea nitrogen and creatinnie values also increased (serum urea nitrogen:9.87+1.27mmol/L to 15.34 2.47mmol/L, creatinnie values:16.08±1.05μmol/L to 31.14±1.81 mol/L), H & E staining showed severe kidney damage. Pretreated QYG could prevent the serum urea nitrogen and creatinine values increased and relieve kidney damage (serum urea nitrogen: 10.28±3.08 mmol/L, creatinnie values:21.01±6.08μmol/L). Compared with the model group mice, QYG can also reduce the accumulation of Pt+in the kidney, downregulated the expression of Ctrl and OCT2, reduced the apoptosis of renal tubular cells, reduced the P53 expression, promoted cell proliferation, reduced mRNA relative expression of TNF-α, COX-2, IL-1β. In the tumor bearing mice model, QYG was found not affect the anti-tumor effect of cisplatin and could reduce the mortality rate of these mice.Conclusion: QYG could effectively reduce cisplatin induced nephrotoxicity, but did not reduce the anti-tumor effect. It could also reduce the mortality of tumor bearing mice. It may be a potential assistant drug of cisplatin in the clinical.