Quantum communication protocols are considered secure provided that all devices included in the system are fully characterized, and side channels are closed. However, as a result of laser radiation exposure, it is possible to change quantum communication systems components’ characteristics. This leads to vulnerabilities appearing in the quantum key distribution system. In this paper, we consider the effect of pulsed laser radiation on fiber-optic isolators used in quantum communication systems. Isolators protect the source of the system from attacking optical radiation coming from the “eavesdropping” side via the quantum channel. Lowering the isolation factor can bring the entire system out of a secure state. This gives an eavesdropper access to information about the secret key. The scenario of the most probable attack to the source of the quantum key distribution system via a pulsed laser was simulated. The experimental setup provided exposure of fiber isolators with pulsed laser radiation at a wavelength of 1064 nm (within the transparency window of the isolators) with a mean power up to 840 mW in four different pulse generation modes. The isolation factor and throughput of tested samples were monitored using a laser diode with a wavelength of 1550 nm and average power of 10.5 mW. Spectrally selective splitters were used to separate the lasers. It is shown that the isolation factor (isolator attenuation in the direction from the quantum channel to the system) at a wavelength of 1550 nm decreases from the initial value of 59.1 dB to 24.5 dB. The throughput (in the direction from the system to the quantum channel) at the same wavelength decreases from 0.6 dB to 1.2–12.3 dB or remains the same, depending on the acting pulsed laser radiation parameters. Temperature monitoring showed that the temperature of the isolator body changes insignificantly when exposed to pulsed radiation. The obtained effects of changing the isolation coefficient and throughput can be explained by the presence of nonlinear effects in the magneto-optical crystal of the isolator. The results of the work can be applicable in the practical evaluation of quantum communication systems security, in particular, the security evaluation of quantum key distribution systems. The results can be used to prepare the standards for certification procedures for assessing the security of quantum communication systems. The work gives recommendations for enhancing signal source unit security in quantum communication systems. Namely, as countermeasures to protect against the effects of pulsed laser radiation, it is proposed to set optical fuses with a limited threshold power, detectors for monitoring input optical radiation power, and narrow-band optical filters at the entrance of the quantum communication system.