| Objective: To explore the effects of intramuscular injection of botulinum toxin typeA(BTX-A)on the expression of pregnant SD rats serum estrogen, progesterone and itsreceptors in the reproductive organs, to understand the toxicity of BTX-Aon embryodevelopment, and to provide theoretical and experimental reference data for BTX-Ain clinical medication safety.Methods:1. The acute toxicity test of botulinum type A: intramuscular injection of BTX-Aon MaleSD rats to detect minimum lethal dose (MLD)and median lethal dose(LD50).(1)chosehealthy adult o-level20male SD rats, randomly divided into4groups, the right side ofthe gastrocnemius muscle was injected of15U/kg,30U/kg,60U/kg,90U/kg WB4doses of BTX-A, and observed the minimal lethal dose of BTX-Aon rats (90U/kg).(2)chose50male SD rats,were randomly divided into5groups, MLD was based on todesign the injection of the right side of the gastrocnemius muscle:62U/kg,77U/kg,96U/kg,120U/kg,150U/kg WB5doses groups, and observed poisoning and death rats ineach group, according to the look-up table method of improved by sun-style synthesis,calculate the LD50of BTX-Aon7days SD rats.2. The effects of BTX-A on the expression of pregnant SD rats serum estrogen, progesterone and its receptors and on embryo development: Chose60pregnant SD rats, rand-omly divided into6groups,10for each group. The negative control group (NS,0.5ml/only)intramuscular injection, Botulinum toxin A group was divided into four doses(BTX-A,30U/kg,60U/kg,90U/kg,120U/kgWB) intramuscular injection and retinoicacid drug control group(100mg/kg WB lavage).Pregnant rats of each group were onday eighth of gestation were administered1times, in pregnancy18drunk from Gastro-dia elata.(1)pregnant rat testing indexes: measuring the pregnancy weight gain rate,the ovarian and uterine weight index, the serum estrogen and progesterone levels using ELISA method, and the expression of reproductive receptor using IHC method.(2)embryos detection indexes: embryo malformation and survival rate, fetal weight, bodylength, tail length and the length of the main bones on limbs, and the detection ofossification.Results:1. The acute toxicity test of botulinum type A:(1) MLD: normal adult male rats weredivided into four groups of intramuscular injection of BTX-A(15U/kg,30U/kg,60U/kg,90U/kg),when the dose was90U/kg,there was the symptoms of poisoning and rats dead,suggesting that MLD of BTX-A in rat is90U/kg.(2) LD50: on the basis of MLD, set upother5dose groups of BTX-A:62U/kg,77U/kg,96U/kg,120U/kg,150U/kg,and thehighest dose group(150U/kg)showed the most severe toxic symptoms, the mortality ratein7days was100%, according to the look-up table method of sun-style synthesis improv-ement,the LD50of BTX-A on rats was114.77U/kg.2The effect of BTX-A on pregnant SD rats body weight gain rate,serum hormones,reproductive organs weight index and hormone receptors: detected pregnant rats of eachgroup at the18thday of gestation:(1)on the15thday of gestation, except for the BTX-A30U/kg dose group, the weight of pregnant rats in other three BTX-A doses groupswere all lower than the negative control group; on the18thday of gestation, only120U/kg dose group weighed less than negative control group, and showed significantdifference (P<0.05).(2) except for the30U/kg dose group, the serum estrogen and proges-terone values of BTX-A4dose groups were all lower than the negative control group,and showed significant difference (P＜0.05).(3) Ovarian and uterine90U/kg,120U/kgdose group weight index were lower than that of the negative control group, and therewas significant difference(P<0.05).(4) Each dose group of BTX-A pregnant rat uterineand ovarian corpus luteum ER, PR expression were higher than that of the negative contr-ol group,there was significant difference (P＜0.05). And enhance with the increase inthe expression of BTX-Adose. 3The effect of BTX-A on the development of in rat embryo E18(1) the BTX-A dose is120U/kg, embryonic live births rate lower than with the negative control group, therewas significant difference (P＜0.05). Embryo deformity appeared in BTX-A60U/kg,90U/kg dose groups.(2)the tail length, embryo weight and body length of the BTX-A120U/kg dose groups were lower than the negative control group, with significant differ-ence (P＜0.05), the placenta weight of the four BTX-Adose groups were lower than thenegative control group, with significant difference (P＜0.05).The upper shoulder blades,humerus, ulna and radius hip and leg length of BTX-A dose60U/kg,90U/kg were lessthan the negative control group, with significant difference (P＜0.05).The length of thefemur, tibia and fibula of BTX-A30U/kg,60U/kg,90U/kg dose were lower than thenegative control group, with significant difference (P＜0.05).The length all bone ossific-ation point in the doses of BTX-A30U/kg,60U/kg,90U/kg were less than negative cont-rol group, with significant difference (P＜0.05).Conclusion:4.1Minimum lethal dose of rat intramuscular injection of BTX-Ais90U/kg, and the med-ian lethal dose is114.77U/kg.4.2Under the conditions of this experiment, when pregnant rats intramuscular injectionof BTX-Adose was less than30U/kg, there was no maternal toxicity, no embryo toxicity.When the dose was greater than60U/kg with the doses increased, pregnant rat embryodevelopmental toxicity and maternal toxicity is aggravated gradually.Tip: In the safe dose range BTX-Atrace intramuscular injection (less than30U/kg)had notoxicity and maternal toxicity in pregnant rats,embryo and teratogenicity. |
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