Glutathione (r-glutamyl cysteingl+glycine, GSH) is a kind ofγ-Amide bond andMercapto groupTripeptide fromglutamate、CysteineandglycineComposition exists in almost every cell of the body[1]。
Glutathione can help maintain normal immune system function, and has antioxidant effect and integrationDetoxification[1]。The sulfhydryl group on cysteine is its active group (often abbreviated as G-SH), which is easy to combine with some drugs, toxins, etc., making it have an integrated detoxification effect[2]。Glutathione can not only be used as medicine, but also as the base material of functional food. It is widely used in functional food such as anti-aging, immunity enhancement, anti-tumor, etc.[2]
Glutathione has two forms: reduced form (G-SH) and oxidized form (G-S-S-G)Reduced glutathioneThe majority.Glutathione reductaseIt can catalyze the tautomerism between the two typescoenzymeIt can also provide NADPH for pentose phosphate bypass metabolism[2]。
Glutathione (GSH) is composed of glutamic acid, cysteine and glycine, and contains sulfhydrylTripeptide, with antioxidant effect and integrationDetoxification。The sulfhydryl group on cysteine is the active group of glutathione (so glutathione is often abbreviated as G-SH), which is easily associated with certain drugs (such asParacetamol）Toxins (such as free radicalsIodoacetic acid、Mustard gas, lead, mercury, arsenic and other heavy metals).Therefore, glutathione (especially glutathione in hepatocytes) can participate in biological transformation, thus transforming harmful toxins in the body into harmless substances and excreting them out of the body.Glutathione also helps maintain normal immune system function[3]。
Content distribution
Announce
edit
Glutathione widely exists in animals and plants, and plays an important role in organisms.It has a high content of 100~1000 mg/100g in baker's yeast, wheat germ and animal liver, 26~34 mg/100g in human blood, 58~73 mg/100g in chicken blood, 10~15 mg/100g in pig blood, high content of 12~33 mg/100g in tomatoes, pineapples and cucumbers, and low content of 0.06~0.7 mg/100g in sweet potatoes, mung bean sprouts, onions and mushrooms[4]。
physiological function
Announce
edit
one
Detoxification: combine with poisons or drugs to eliminate their toxic effects[5]；
two
Participate in redox reaction: as an important reducing agent, participate in various redox reactions in the body[5]；
three
Protect the activity of sulfhydrylase: keep SH, the active group of sulfhydrylase, in a reduced state[5]；
four
Maintain the stability of erythrocyte membrane structure: eliminate the destruction of oxidants on erythrocyte membrane structure[5]。
Action mechanism
Announce
edit
As an important intracellular metabolic regulator, GSH is not only the cofactor of glyceraldehyde phosphate dehydrogenase, but also the cofactor of glyoxalase and triose dehydrogenase. It participates in the tricarboxylic acid cycle and sugar metabolism in the body, and can activate a variety of enzymes, such as sulfhydryl (SH) enzyme coenzyme, to promote the metabolism of sugars, fats and proteins.GSH molecule is characterized by active sulfhydryl group (- SH), which is the most important functional group. It can participate in many important biochemical reactions in the body, protect the sulfhydryl group of important enzymes in the body from oxidation and inactivation, and ensure energy metabolism and cell utilization.At the same time, it can directly reduce free radicals into acidic substances through the combination of sulfhydryl group and free radicals in the body, thus accelerating the excretion of free radicals and combating the damage of free radicals to important organs.Haddad et al. found that GSH was involved in the regulation of lipopolysaccharide induced cytokine transcription and I-KB/NF-KB signaling pathway.Armstrong et al found that the decrease of GSH content is a potential early activation signal of apoptosis, and the subsequent oxygen free radicals promote cell apoptosis[6]。
Main applications
Announce
edit
Clinical drugs
Glutathione drug is widely used in clinic. In addition to using its sulfhydryl group to chelate heavy metals, fluoride, mustard gas and other toxins, it is also used in hepatitis, hemolytic diseases, keratitiscataractAnd retinal diseases, as a treatment or adjuvant treatment of drugs.In recent years, Western scientists, especiallyJapanScholars found that glutathione has the function of inhibiting HIV[7]。
The latest research also shows that GSH can correct the imbalance of acetylcholine and cholinesterase, play an anti allergic role, prevent skin aging and pigmentation, reduce the formation of melanin, improve the skin's antioxidant capacity and make the skin shiny. In addition, GSH also has a very good role in the treatment of corneal diseases and improving sexual function[7]。
Antioxidant effect
As an important antioxidant in the body, glutathione can eliminate free radicals in the body;Because GSH itself is easily oxidized by some substances, it can protect the sulfhydryl groups in many proteins, enzymes and other molecules from being oxidized by harmful substances in the body, thus ensuring the normal functioning of proteins, enzymes and other molecular physiological functions;There is a lot of glutathione in human red blood cells, which is of great significance in protecting the sulfhydryl group of protein on the red blood cell membrane from being reduced and preventing hemolysis[7]。
food additives
Adding glutathione to food can play an unexpected role:
1. When added to flour products, it can play a reducing role.It not only shortens the time for making bread to one half or one third of the original time, greatly improves the working conditions, and plays a role in strengthening food nutrition and other functions[8]。
2. When added to yogurt and infant food, it is equivalent to vitamin C and can act as a stabilizer[8]。
3. Stir it in the fish cake to prevent the color from deepening[8]。
4. When added to meat products, cheese and other foods, it has the effect of enhancing flavor[8]。
Detection means
Announce
edit
There are many methods to detect glutathione, includingIodometry、DTNB(Ellman reagent)method、Sodium nitroso ferricyanide method、Fluorescence method、Alloxan methodandHigh performance liquid chromatography（ HPLC）。Each of the above methods has its own advantages. When detecting GSH content, a suitable, sensitive and accurate detection method can be selected according to the actual situation[9]。
DTNB colorimetry
Principle:The thiol reaction of DTNB with glutathione produces yellow 5-thio-2-nitrobenzoic acid, which has a maximum absorption peak at 412 nm, while DTNB has almost no absorption peak above 400 nm.Add a small amount of DTNB to the sample containing glutathione, measure the absorption value of the reaction solution at 412 nm, and calculate the glutathione content according to the absorption value[9]。
Method steps:
1. Preparation of DTNB storage solution: dissolve 0.01 mol/L DTNB in 0.05 mol/L phosphoric acid buffer (pH7.0) to form DTNB storage solution[9]。
2. Preparation of DTNB analytical solution: dilute DTNB storage solution 100 times with Tris HCl buffer of 0.5 mol/L and pH 8.0 to prepare DTNB analytical solution, place it away from light, and prepare it for use[9]。
3. Operation steps: Take 0.5 mL of glutathione standard solution or sample solution to be tested (0.1~3.0 mol/L), add it to 1.5 mL of 0.15 mol/L NaOH solution, add 0.5 mL of 3% formaldehyde solution, and react at pH 8.0 and 25 ℃ for 2 minutes. After the reaction, take 0.5 mL of reaction solution and add it to DTNB analytical solution, react at 25 ℃ for 5 minutes, and determine the absorption value at 412 nm.Calculate the difference according to the absorbance of the two and substitute it into the standard curve to calculate the content of glutathione[9]。
Iodometry
Principle:Use the reducibility of glutathione to react with potassium iodate. When glutathione completely reacts, potassium iodate oxidizes potassium iodide into iodine, and iodine turns the starch indicator blue, which is the titration end point[9]。
Operation steps: Accurately weigh 0.05 mg of glutathione sample, put it into a 10 mL beaker, and use 2% HPOthreeDissolve, transfer to a 100 mL volumetric flask, and dilute to 100 mL with water.Suck 5mL of the above solution into a 100mL triangular flask, and then add 2% HPOthree5 mL of solution is made into 10 mL, and then 1 mL of 5% potassium iodide and 2 drops of starch indicator are added into it.Use 0.001 mol/L potassium iodate for titration. When the consumption value of 0.001 mol/L potassium iodate is consistent with the theoretical value of 2.445 mL, the content of glutathione in the finished product is 100%[9]。
Calculation formula:
, where V is the volume of 0.001 mol/L potassium iodate consumed (mL), and the volume of 2.445 - 100% content should be consumed (mL)[9]。
High performance liquid chromatography
principle: High performance liquid chromatography is a process of continuous separation of solutes due to different partition coefficient, affinity, molecular size, adsorption capacity, etc. between the same stationary phase and mobile phase[9]。
Operation steps：
① Chromatographic conditions
Chromatographic column: Kromasil C18 column (4.6 mm × 250 mm, 5μm）[9]。
Mobile phase: mixed solution of sodium dihydrogen phosphate and sodium octanesulfonate (3.0 g of sodium dihydrogen phosphate and 1.0 g of sodium octanesulfonate, dissolved with water and fixed to 500 mL, adjusted with phosphoric acid to pH 3): acetonitrile=96:4 (volume ratio)[9]。
Accurately weigh an appropriate amount of glutathione standard, dissolve it in a volumetric flask with deionized water, mix it evenly and make it into 25μg/mL、50 μg/mL、100 Vg/mL、200 μg/mL、400 μg/mL、800 μG/mL series concentration sample solution as standard solution[9]。
③ Sample preparation
Accurately weigh the glutathione sample, add deionized water, mix well, and make the concentration of 25~800μG/mL sample solution as analytical solution[9]。
④ Draw standard curve
Suck 10% of the prepared glutathione standard solution respectivelyμL sample injection measurement, draw the standard curve with the concentration and peak area of glutathione and glutathione production technology, and obtain the regression equation of the curve according to the standard curve.The change of peak area should have a linear relationship with the mass concentration of glutathione, and its fitting degree should be above 0.9995[9]。
⑤ Determination of samples
Take glutathione sample analysis solution 10μ50. Use the high performance liquid chromatograph to measure the sample injection, get the peak area of the sample solution, and calculate the content of glutathione according to the regression equation[9]。
Glutathione is a tripeptide compound containing sulfhydryl group, which has important physiological activities in human body, such as activating redox system, activating enzymes, detoxification and so on.Glutathione exists in the body in two forms: reduced and oxidized. Its active component is reduced glutathione, which participates in the tricarboxylic acid cycle and sugar metabolism in the body, promotes the production of high energy in the body, and plays the role of coenzyme.Reduced glutathione is the coenzyme of glyceraldehyde phosphate deoxygenase, glyoxalase and triose phosphate deaminase.Reduced glutathione can activate SH enzyme in the body, promote the metabolism of carbohydrate, fat and protein, and regulate the metabolic process of cell membrane.Reduced glutathione can combine with a variety of exogenous and endogenous toxic substances to produce detoxification substances[10]。
Indications
This product is used for adjuvant treatment of chronic liver diseases.It includes liver damage caused by virus, drug toxicity and alcohol toxicity. It can also be used for poisoning of organic phosphorus, amino or nitro aromatic compounds[10]。
Usage and dosage
Oral
It is placed between the buccal mucosa and the gingiva. For adults, 0.4 g (4 tablets) is taken 3 times a day. Generally, 12 weeks is a course of treatment[10]。
Intramuscular and intravenous administration
(1) It can be used as an auxiliary drug for chemotherapy (cisplatin, cyclophosphamide, adriamycin, daunorubicin, bleomycin), which can reduce the damage caused by chemotherapy without affecting the efficacy, thus increasing the dose of chemotherapy[10]。
First dose 1500 mg/mtwo, dissolved in 100 ml of normal saline or 5% glucose injection, intravenous infusion within 15 minutes;On the second to fifth days, 600 mg was injected intramuscularly every day.After cyclophosphamide treatment, intravenous infusion for 15 minutes should be carried out immediately to reduce the impact of chemotherapy on the urinary system.For cisplatin treatment, the dose of reduced glutathione should not exceed 35mg/mg cisplatin, so as not to affect chemotherapy or follow the doctor's advice[10]。
(2) It can be used for auxiliary treatment of liver injury caused by alcohol, virus, drugs and other chemicals.
① For viral hepatitis, 1200 mg, once a day, intravenous drip, 30 days; ②Severe hepatitis, 1200~2400 mg, once a day, intravenous drip, 30 days; ③Active cirrhosis, 1200mg, once a day, intravenous drip, 30 days; ④Fatty liver, 1800 mg, once a day, intravenous drip, 30 days; ⑤Alcoholic hepatitis, 1800 mg, once a day, intravenous drip, 14-30 days; ⑥Drug hepatitis, 1200~1800 mg, once a day, intravenous drip, 14~30 days[10]。
(3) It is used as adjuvant medicine for radiotherapy, and is administered after irradiation, with a dose of 1500 mg/mtwo, or as directed by your doctor[10]。
(4) It can be used for the treatment of hypoxemia. The dose is 500 mg/m, dissolved in 100 ml of normal saline, administered intravenously, and maintained by 300~600 mg intramuscular injection every day[10]。
(5) Note for use: (300-600 mg) must be completely dissolved in the solution when intramuscular injection, and the solution must be clear and colorless.For intravenous administration, the drug can be dissolved by the lysate and then injected slowly. At least 20 ml of lysate is required for intravenous drip administration[10]。
Adverse reactions
1. Mild oral leukoplakia, ulcer, tongue coating peeling and pain and other oral discomfort[10]。
2. Rare sudden rash[10]。
3. Occasionally, they suffer from anorexia, nausea, vomiting, epigastric pain, etc.It disappeared after drug withdrawal, and the injection site was slightly painful[10]。
matters needing attention
1. It is recommended to discontinue this product in case of adverse oral reaction[10]。
2. Keep away from children[10]。
3. Use with caution for patients with phenylketonuria[10]。
4. The dissolved solution shall be used immediately, and the remaining solution shall not be reused[10]。
5. Intramuscular injection is only used when this route is needed, and repeated injections at the same site should be avoided[10]。
